gov.noaa.nodc:0294041 eng utf8 dataset NOAA National Centers for Environmental Information ncei.info@noaa.gov https://www.ncei.noaa.gov/contact HTTPS NOAA Contact Information Information for contacts at NCEI. information custodian 2026-07-10T20:41:12Z ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data ISO 19115-2:2009(E) ModMon Neuse River Estuary and Pamlico Sound Water Quality Monitoring from 1994-01-24 to 2022-01-22 (NCEI Accession 0294041) 2025-01-27 publication 2025-01-27 creation NOAA National Centers for Environmental Information gov.noaa.nodc:0294041 NCEI Archive Management System 0294041 NOAA National Centers for Environmental Information ncei.info@noaa.gov https://www.ncei.noaa.gov/contact HTTPS NCEI Contact Information Information for contacts at NCEI. information publisher Chris Turner Alaska Ocean Observing System (AOOS) chris@axiomalaska.com http://www.aoos.org HTTP Alaska Ocean Observing System (AOOS) website Institution web page information resourceProvider Axiom Data Science https://www.axiomdatascience.com/ HTTPS Axiom Data Science website Institution web page information resourceProvider Nathan Hall University of North Carolina - Chapel Hill (UNC) nshall@email.unc.edu principalInvestigator Hall, Nathan University of North Carolina - Chapel Hill author Paerl, Hans author Rossignol, Karen author Sloup, Randy author University of North Carolina - Chapel Hill (UNC) contributor tableDigital The Neuse River Estuary Water Quality Dataset is a compilation of the biological, chemical and physical water quality data that was collected along the length of the Neuse River Estuary, North Carolina from January 24, 1994 to the present. The primary purpose of this dataset was to provide long-term environmental information to supplement experimental, process-based research, including the Atlantic Coast Environmental Indicators Consortium (ACE-INC) project as well as other laboratory studies. This dataset is composed of four spreadsheets (xlsx) containing regular water sampling and in situ measurements collected at fixed sampling stations in the Neuse River and Pamlico Sound. The data in the files titled "SECOORA_NRWQ..." and SECOORA_PSWQ..." contain water samples and in situ measurements collected at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). The data in the file titled "SECOORA_NR_YSI..." and "SECOORA_PS_YSI..." contain in situ measurements performed throughout the water column in 0.5 meter depth increments. Parameters measured include: temperature, salinity, specific conductivity, dissolved oxygen (DO), pH, chlorophyll fluorescence, photosynthetically active radiation (PAR), turbidity, barometric pressure, Secchi depth, colored dissolved organic matter (CDOM), particulate organic carbon (POC) and nitrogen (PN), dissolved organic and inorganic carbon, dissolved inorganic nutrient concentrations (nitrate/nitrite, ammonium, total dissolved nitrogen, phosphate and silicic acid), chlorophyll a, primary productivity and diagnostic phytoplankton pigment concentrations (chlorophylls and carotenoids). Calculated parameters include: diffuse light attenuation coefficient (Kd), carbon to nitrogen molar ratio (C:N), dissolved inorganic nitrogen (DIN; nitrate/nitrite plus ammonium), dissolved organic nitrogen (DON; total dissolved nitrogen minus dissolved inorganic nitrogen) and the nitrogen to phosphorus molar ratio (N:P). The Neuse River Estuary Modeling and Monitoring Project (ModMon) is a collaborative effort between the University of North Carolina and the North Carolina Department of Environment and Natural Resources. The ModMon project provides high quality, readily available data on water quality parameters in the Neuse River. These parameters can be used to evaluate the overall health of the river and address the modeling and monitoring needs for the Neuse River Estuary Total Maximum Daily Load (TMDL). The data collected are available on a continuous basis to government agencies and universities. ModMon also supports numerous undergraduate field trip experiences for universities. pending NOAA National Centers for Environmental Information ncei.info@noaa.gov https://www.ncei.noaa.gov/contact HTTPS NCEI Contact Information Information for contacts at NCEI. information pointOfContact asNeeded https://www.ncei.noaa.gov/access/metadata/landing-page/bin/gfx?id=gov.noaa.nodc:0294041 Graphic not available. PNG 0294041 NCEI ACCESSION NUMBER 2024-04-03 publication AMMONIUM (NH4) BAROMETRIC PRESSURE CHLOROPHYLL CHLOROPHYLL A - INTEGRATED CHLOROPHYLL A CONCENTRATION Chromophoric Dissolved Organic Matter (CDOM) CONDUCTIVITY DEPTH - OBSERVATION Diadinoxanthin diatom DISSOLVED INORGANIC CARBON (DIC) DISSOLVED ORGANIC CARBON Dissolved Organic Nitrogen DISSOLVED OXYGEN fucoxanthin LIGHT ATTENUATION monadoxanthin nitrate + nitrite content (concentration) NITROGEN - PARTICULATE PARTICULATE ORGANIC CARBON PERCENT OXYGEN pH PHOTOSYNTHETIC ACTIVE RADIATION (PAR) PRIMARY PRODUCTIVITY SALINITY Secchi depth silica suspended solids Total Dissolved Nitrogen (TDN) turbidity Violaxanthin WATER TEMPERATURE theme NODC DATA TYPES THESAURUS bottle flow injection analyzer fluorometer oxygen sensor PAR Sensor pH sensor Secchi disk thermometer Total Organic Carbon (TOC) analyzer turbidity sensor instrument NODC INSTRUMENT TYPES THESAURUS in situ laboratory analyses theme NODC OBSERVATION TYPES THESAURUS University of North Carolina - Chapel Hill dataCentre NODC COLLECTING INSTITUTION NAMES THESAURUS Axiom Data Science dataCentre NODC SUBMITTING INSTITUTION NAMES THESAURUS Southeast Coastal Ocean Observing Regional Association (SECOORA) project NODC PROJECT NAMES THESAURUS North Atlantic Ocean Pamlico Sound place NODC SEA AREA NAMES THESAURUS oceanography theme WMO_CategoryCode 2012-09-15 publication Axiom Data Science dataCentre Global Change Master Directory (GCMD) Data Center Keywords 2026 revision 24 Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA) Greenbelt MD https://forum.earthdata.nasa.gov/app.php/tag/GCMD%2BKeywords HTTPS GCMD Keyword Forum Page Global Change Master Directory (GCMD). 2025. GCMD Keywords, Version 21. Greenbelt, MD: Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA). URL (GCMD Keyword Forum Page): https://forum.earthdata.nasa.gov/app.php/tag/GCMD+Keywords information custodian IOOS > Integrated Ocean Observing System project Global Change Master Directory (GCMD) Project Keywords 2026 revision 24 Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA) Greenbelt MD https://forum.earthdata.nasa.gov/app.php/tag/GCMD%2BKeywords HTTPS GCMD Keyword Forum Page Global Change Master Directory (GCMD). 2025. GCMD Keywords, Version 21. Greenbelt, MD: Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA). URL (GCMD Keyword Forum Page): https://forum.earthdata.nasa.gov/app.php/tag/GCMD+Keywords information custodian EARTH SCIENCE > ATMOSPHERE > ATMOSPHERIC PRESSURE EARTH SCIENCE > BIOLOGICAL CLASSIFICATION > PLANTS > MICROALGAE > DIATOMS EARTH SCIENCE > BIOSPHERE > ECOLOGICAL DYNAMICS > ECOSYSTEM FUNCTIONS > PRIMARY PRODUCTION EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > AMMONIA EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > CHLOROPHYLL EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > INORGANIC CARBON EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > NITRATE EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > NITRITE EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > NITROGEN EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > ORGANIC CARBON EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > ORGANIC MATTER EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > OXYGEN EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > PH EARTH SCIENCE > OCEANS > OCEAN CHEMISTRY > SUSPENDED SOLIDS EARTH SCIENCE > OCEANS > OCEAN OPTICS > ATTENUATION/TRANSMISSION EARTH SCIENCE > OCEANS > OCEAN OPTICS > PHOTOSYNTHETICALLY ACTIVE RADIATION EARTH SCIENCE > OCEANS > OCEAN OPTICS > SECCHI DEPTH EARTH SCIENCE > OCEANS > OCEAN OPTICS > TURBIDITY EARTH SCIENCE > OCEANS > OCEAN TEMPERATURE > WATER TEMPERATURE EARTH SCIENCE > OCEANS > SALINITY/DENSITY > CONDUCTIVITY EARTH SCIENCE > OCEANS > SALINITY/DENSITY > SALINITY theme Global Change Master Directory (GCMD) Science Keywords 2026 revision 24 Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA) Greenbelt MD https://forum.earthdata.nasa.gov/app.php/tag/GCMD%2BKeywords HTTPS GCMD Keyword Forum Page Global Change Master Directory (GCMD). 2025. GCMD Keywords, Version 21. Greenbelt, MD: Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA). URL (GCMD Keyword Forum Page): https://forum.earthdata.nasa.gov/app.php/tag/GCMD+Keywords information custodian 19'-Butanoyloxyfucoxanthin 19'-Hexanoyloxyfucoxanthin 9'-cis Neoxanthin Carbon to nitrogen molar ratio Chlorophyllide a Corrected Chromophoric Dissolved Organic Matter (CDOM) Cryptophytes Diffuse light attenuation coefficient Dinoflagellates Dissolved inorganic nitrogen Lutein Myxoxanthophyll PAR Depth PHOTOSYNTHETIC ACTIVE RADIATION (PAR) - Channel 1 PHOTOSYNTHETIC ACTIVE RADIATION (PAR) - Channel 2 Peridinin Pheophorbide a Prasinoxanthin Total CHLOROPHYLL A Total suspended solids YSI Temperature YSI Time YSI barometric pressure YSI chlorophyll concentration YSI chlorophyll raw fluorescence YSI depth YSI dissolved oxygen YSI dissolved oxygen saturation YSI ph YSI salinity YSI specific conductivity YSI turbidity Zeaxanthin alloxanthin antherexanthin assimilation index beta-carotene canthaxanthin chlorophyll a corrected chlorophyll c1 and c2 chlorophytes cyanobacteria diatoxanthin divinyl chlorophyll b echinenone gyroxanthin nitrogen to phosphorus molar ratio orthophosphate pheophytin a uncorrected Chromophoric Dissolved Organic Matter (CDOM) theme Provider Keywords FIA > FLOW INJECTION ANALYSIS FLUOROMETERS > FLUOROMETERS OXYGEN METERS > OXYGEN METERS PAR SENSORS > Photosynthetically Active Radiation Sensors PH METERS > PH METERS SECCHI DISKS > SECCHI DISKS THERMOMETERS > THERMOMETERS TOC > Total Organic Carbon Analyzer TURBIDITY METERS > TURBIDITY METERS instrument Global Change Master Directory (GCMD) Instrument Keywords 2026 revision 24 Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA) Greenbelt MD https://forum.earthdata.nasa.gov/app.php/tag/GCMD%2BKeywords HTTPS GCMD Keyword Forum Page Global Change Master Directory (GCMD). 2025. GCMD Keywords, Version 21. Greenbelt, MD: Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA). URL (GCMD Keyword Forum Page): https://forum.earthdata.nasa.gov/app.php/tag/GCMD+Keywords information custodian UWO-PAR 6067 YSI - handheld multi-parameter instrument field light simulator glass fiber filters integrated water sampler spherical underwater quantum sensor instrument Provider Instruments OCEAN > ATLANTIC OCEAN > NORTH ATLANTIC OCEAN place Global Change Master Directory (GCMD) Location Keywords 2026 revision 24 Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA) Greenbelt MD https://forum.earthdata.nasa.gov/app.php/tag/GCMD%2BKeywords HTTPS GCMD Keyword Forum Page Global Change Master Directory (GCMD). 2025. GCMD Keywords, Version 21. Greenbelt, MD: Earth Science Data and Information System, Earth Science Projects Division, Goddard Space Flight Center (GSFC), National Aeronautics and Space Administration (NASA). URL (GCMD Keyword Forum Page): https://forum.earthdata.nasa.gov/app.php/tag/GCMD+Keywords information custodian 9440GJ Send2NCEI Submission Package ID Data in this archival information package are not currently available for public download from NCEI. Please contact NCEI directly for access to these data. accessLevel: restricted public otherRestrictions Cite as: Hall, Nathan; Paerl, Hans; Rossignol, Karen; Sloup, Randy (2025). ModMon Neuse River Estuary and Pamlico Sound Water Quality Monitoring from 1994-01-24 to 2022-01-22 (NCEI Accession 0294041). [indicate subset used]. NOAA National Centers for Environmental Information. Unpublished Dataset. https://www.ncei.noaa.gov/archive/accession/0294041. Accessed [date]. Distribution liability: NOAA and NCEI make no warranty, expressed or implied, regarding these data, nor does the fact of distribution constitute such a warranty. NOAA and NCEI cannot assume liability for any damages caused by any errors or omissions in these data. If appropriate, NCEI can only certify that the data it distributes are an authentic copy of the records that were accepted for inclusion in the NCEI archives. otherRestrictions Use liability: NOAA and NCEI cannot provide any warranty as to the accuracy, reliability, or completeness of furnished data. Users assume responsibility to determine the usability of these data. The user is responsible for the results of any application of this data for other than its intended purpose. eng; USA utf8 environment oceans climatologyMeteorologyAtmosphere -77.2 -76.2 34.9 35.3 1994-01-24 2022-01-22 NOAA National Centers for Environmental Information +1-301-713-3277 ncei.info@noaa.gov pointOfContact In most cases, electronic downloads of the data are free. However, fees may apply for custom orders, data certifications, copies of analog materials, and data distribution on physical media. Data in this archival information package are not currently available for public download from NCEI. Please contact NCEI directly for access to these data. Originator data format https://www.ncei.noaa.gov/archive/accession/0294041 HTTPS Web browser NCEI Dataset Landing Page Navigate directly to the URL for a descriptive web page with download links. information https://www.ncei.noaa.gov/archive/accession/oas/294041 HTTPS Web browser Descriptive Information Navigate directly to the URL for a descriptive web page. offlineAccess https://www.ncei.noaa.gov/archive/accession/download/294041 HTTPS Web browser HTTPS Navigate directly to the URL for a descriptive web page. offlineAccess https://www.ncei.noaa.gov/contact HTTPS NOAA National Centers for Environmental Information Users may contact NCEI for more information and to request access to the archived data. User requests must be approved by NCEI and are subject to order fulfillment constraints. offlineAccess dataset Parameter or Variable: YSI Temperature (measured); Units: degrees C; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI depth (measured); Units: meter; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI specific conductivity (measured); Units: millisiemen/centimeter; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI salinity (calculated); Units: ppt; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI dissolved oxygen (measured); Units: ppt; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI dissolved oxygen saturation (measured); Units: percent; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI ph (measured); Units: pH; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI turbidity (measured); Units: NTU; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI chlorophyll raw fluorescence (measured); Units: relative fluorescence units; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI chlorophyll concentration (measured); Units: microgram/liter; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: YSI barometric pressure (measured); Units: millimeters Hg; Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument; Sampling and Analyzing Method: In situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, EDS-S or V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561), a YSI dissolved oxygen probe (Model 6562 or 6150), a YSI turbidity probe (Model 6136), and a LI-COR underwater quantum sensor for PAR (LI-192SA). The YSI sonde was coupled to a YSI 650 MDS datalogger. In situ measurements were performed at the surface (approximately 0.2 meters) and at the bottom of the water column (approximately 0.5 meters from the sediment layer). In situ measurements were also performed throughout the water column in 0.5 meter depth increments. These data are included in the table labeled profile data. The data were stored on the datalogger and downloaded using EcoWatch (YSI) software upon return to the laboratory. (Calibration info needed) Parameter or Variable: Secchi depth (measured); Units: meters per second; Observation Category: in situ; Sampling Instrument: Secchi disk; Sampling and Analyzing Method: The secchi disk was deployed off of the sunlit side of the research vessel. The depth (in meters) at which the secchi disk was no longer visible by the naked eye was recorded as the secchi depth. Parameter or Variable: Diffuse light attenuation coefficient (calculated); Units: per meter; Observation Category: in situ; Sampling Instrument: spherical underwater quantum sensor; Sampling and Analyzing Method: The diffuse light attenuation coefficient, Kd, was calculated from depth profiles of photosynthetically active radiation (PAR, 400-700 nm). Prior to the 07/30/2003 sampling date, PAR measurements were performed with a spherical underwater quantum sensor (LI-COR LI-193SA) coupled to a LI-COR LI-1000 datalogger. Beginning on the 07/30/2003 sampling date, a flat underwater quantum sensor (LI-COR LI-193SA) attached to a Yellow Springs Instruments YSI 6600 or YSI 6600 EDS-S sonde was used to measure PAR. Measurements of PAR were performed on the sunlit side of the research vessel in 0.5 meter depth increments, beginning just below the water surface. The diffuse attenuation coefficient is the slope of the linear regression between natural log transformed PAR data and depth. Parameter or Variable: Total suspended solids (measured); Units: milligram/liter; Observation Category: in situ; Sampling Instrument: glass fiber filters; Sampling and Analyzing Method: Gravimetric analysis of seston collected on GF/F glass fiber filters. Analysis performed by Lois Kelly for John Wells. Parameter or Variable: PARTICULATE ORGANIC CARBON (measured); Units: micrograms of carbon per liter; Observation Category: laboratory analysis; Sampling Instrument: glass fiber filters; Sampling and Analyzing Method: Particulate organic carbon (POC) concentrations were determined by elemental analysis of material collected on pre-combusted Whatman GF/F glass fiber filters. Carbonates were removed from the filters by vapor phase acidification using concentrated hydrochloric acid (HCl). After drying at 60 0C, the filters were rolled in tin disks and injected into a PE 2400 Series II CHNS/O Analyzer calibrated with acetanilide ending in June 2014. Starting on the Pamlico Sound sample date of June 3, 2014, a Costech Analytical Technologies, Inc. Elemental Combustion System CHNS-O ECS 4010 was used for elemental analysis by "flash combustion/chromatographic separation and multi-detector techniques". The Costech Instrument utilizes EAS Clarity Software. Atropine standards are used to develop a calibration curve (C 70.56%, N 4.84%, and carbon response ratio of 0.025 +/-0.003). NIST Buffalo River Sediment Reference Material 8704 (C 3.351% +/-0.017, N 0.20% +/-0.04) and/or Acetanilide Bypass (C 71.09%, N 10.36%, carbon response ratio of 0.055 +/- 0.003) may used for calibration or a check standard. Parameter or Variable: NITROGEN - PARTICULATE (measured); Units: micrograms of nitrogen per liter; Observation Category: laboratory analysis; Sampling Instrument: glass fiber filters; Sampling and Analyzing Method: Particulate Nitrogen (PN) concentrations were determined by elemental analysis of material collected on pre-combusted Whatman GF/F glass fiber filters. Carbonates were removed from the filters by vapor phase acidification using concentrated hydrochloric acid (HCl). After drying at 60 0C, the filters were rolled in tin disks and injected into a PE 2400 Series II CHNS/O Analyzer calibrated with acetanilide ending in June 2014. Starting on the Pamlico Sound sample date of June 3, 2014, a Costech Analytical Technologies, Inc. Elemental Combustion System CHNS-O ECS 4010 was used for elemental analysis by "flash combustion/chromatographic separation and multi-detector techniques". The Costech Instrument utilizes EAS Clarity Software. Atropine standards are used to develop a calibration curve (C 70.56%, N 4.84%, and carbon response ratio of 0.025 +/-0.003). NIST Buffalo River Sediment Reference Material 8704 (C 3.351% +/-0.017, N 0.20% +/-0.04) and/or Acetanilide Bypass (C 71.09%, N 10.36%, carbon response ratio of 0.055 +/- 0.003) may used for calibration or a check standard. Parameter or Variable: Carbon to nitrogen molar ratio (calculated); Units: unitless; Observation Category: laboratory analysis; Sampling Instrument: none; Sampling and Analyzing Method: (Carbon ug/L divided by 12.011)/(Nitrogen ug/L divided by 14.007) Parameter or Variable: DISSOLVED ORGANIC CARBON (measured); Units: micromole/liter; Observation Category: laboratory analysis; Sampling Instrument: Total Organic Carbon (TOC) Analyzer; Sampling and Analyzing Method: Dissolved organic carbon (DOC) was measured after vacuum filtration (< 25 kPA) of the collected water samples through pre-combusted (3-4 hours at 450 0C) Whatman GF/F glass fiber filters. The filtrate was stored in pre-combusted glass scintillation vials with Teflon closures and frozen (-20 0C) until analysis. Dissolved organic carbon concentrations were measured using a Shimadzu TOC-5000A Analyzer. This instrument uses high temperature catalytic oxidation followed by non-dispersive infrared analysis of the CO2 produced. Samples were acidified to pH < 2 and sparged with air before they were analyzed for non-volatile organic carbon. DOC values in 1996 were run from previously run nutrient samples. Parameter or Variable: DISSOLVED INORGANIC CARBON (DIC) (measured); Units: milligram/liter; Observation Category: laboratory analysis; Sampling Instrument: Total Organic Carbon (TOC) Analyzer; Sampling and Analyzing Method: Dissolved inorganic carbon was measured by acidification followed by infrared analysis of carbon dioxide (CO2). Shimadzu Total Organic Carbon Analyzer (TOC-5000A). NR ModMon samples from 5/10/2018 run in duplicate resulted in standard error analysis measurement (RSD) of DIC +/- 0.94%. Parameter or Variable: nitrate + nitrite content (concentration) (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: flow injection analyzer; Sampling and Analyzing Method: Nitrate plus nitrite was measured after vacuum filtration (< 25 kPA) of the collected water samples through pre-combusted (3-4 hours at 450 0C) Whatman GF/F glass fiber filters. The filtrate was stored in high-density polyethylene bottles and frozen (-20 0C) until analysis. Two replicates were run from the same sample bottle. Nitrate plus nitrite concentrations were determined using a Lachat Quick Chem 8000 flow injection autoanalyzer (Milwaukee, WI, USA) using method FIA 31-107-04-1-C. The method detection limit from the 10/6/99 through 10/18/02 sampling dates was 1.06 µg L-1. The method detection limit beginning on the 11/21/02 sampling date was 3.68 µg L-1. The mdl starting on sampling date of 10/24/2014 was 0.36 µg L-1. Mdl on sampling date of 2/4/2015 was changed to 0.71 µg L-1. MDL on sampling date of 8/14/2017 was changed to 0.88 µg L-1. MDL 0.71 ug/L on 1/25/2020. Parameter or Variable: AMMONIUM (NH4) (measured); Units: micrograms Nitrogen per liter; Observation Category: laboratory analysis; Sampling Instrument: flow injection analyzer; Sampling and Analyzing Method: Ammonium (NH4+) was measured after vacuum filtration (< 25 kPA) of the collected water samples through pre-combusted (3-4 hours at 450 0C) Whatman GF/F glass fiber filters. The filtrate was stored in high-density polyethylene bottles and frozen (-20 0C) until analysis. Two replicates were run from the same sample bottle. Concentrations were determined using a Lachat Quick Chem 8000 flow injection autoanalyzer (Milwaukee, WI, USA) using method FIA 31-107-06-1-A/B. The method detection limit from the 10/6/99 through 10/18/02 sampling dates was 4.69 µg L-1. The method detection limit beginning on the 11/21/02 sampling date was 4.31 µg L-1.The mdl starting on sampling date of 10/24/2014 was 2.87 µg L-1. Mdl on sampling date of 2/4/2015 was changed to 3.34 µg L-1. MDL on sampling date of 8/14/2017 was changed to 1.05 µg L-1. MDL of 6.99 on 1/25/2020. Parameter or Variable: Dissolved inorganic nitrogen (calculated); Units: micrograms of nitrogen per liter; Observation Category: laboratory analysis; Sampling Instrument: none; Sampling and Analyzing Method: Dissolved inorganic nitrogen (DIN) concentration was calculated by summing nitrate/nitrite (NO3- / NO2-) and ammonium (NH4+). If either NO3- / NO2- or NH4+ were below the detection limit (-9999), they were taken to be zero for this calculation. Parameter or Variable: Total Dissolved Nitrogen (TDN) (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: flow injection analyzer; Sampling and Analyzing Method: Total dissolved nitrogen (TDN) was measured using the Lachat Quick Chem 8000 flow injection autoanalyzer (Milwaukee, WI, USA) using method FIA 31-107-04-3-B. TDN 1999, 2000 and 2001 samples were run in May 2004. These values are questionable as the samples were stored for a long time, opened and unopened and were unfrozen and refrozen. The mdl starting on sampling date of 10/24/2014 was 19.6 µg L-1. Mdl on sampling date of 2/4/2015 was changed to 10.5 µg L-1. MDL on sampling date of 8/14/2017 was changed to 7.30 µg L-1. MDL of 11.75 on 1/25/2020 Parameter or Variable: Dissolved Organic Nitrogen (calculated); Units: microgram/liter; Observation Category: other; Sampling Instrument: none; Sampling and Analyzing Method: Dissolved organic nitrogen (DON) was calculated by subtracting dissolved inorganic nitrogen (DIN) from total dissolved nitrogen (TDN). If the DIN value used in the calculation was below the detection limit, it was take to be zero for this calculation. At one point DON was determined by high temperature oxidation using the Antek 7000N or Antek 7000V analyzer. Parameter or Variable: orthophosphate (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: flow injection analyzer; Sampling and Analyzing Method: Orthophosphate (PO43-) was measured after vacuum filtration (< 25 kPA) of the collected water samples through pre-combusted (3-4 hours at 450 0C) Whatman GF/F glass fiber filters. The filtrate was stored in high-density polyethylene bottles and frozen (-20 0C) until analysis. Two replicates were run from the same sample bottle. Concentrations were determined using a Lachat Quick Chem 8000 flow injection autoanalyzer (Milwaukee, WI, USA) using method FIA 31-115-01-1-F/G. The method detection limit from the 10/6/99 through 11/21/02 sampling dates was 0.35 µg L-1. The method detection limit beginning on the 11/21/02 sampling date was 0.74 µg L-1. The mdl starting on sampling date of 10/24/2014 was 0.69 µg L-1. Mdl on sampling date of 2/4/2015 was changed to 0.61 µg L-1. MDL on sampling date of 8/14/2017 was changed to 1.80 µg L-1. MDL of 6.56 on 1/25/2020. Parameter or Variable: silica (measured); Units: micromole/liter; Observation Category: laboratory analysis; Sampling Instrument: flow injection analyzer; Sampling and Analyzing Method: Silicic acid (SiO2) was measured after vacuum filtration (< 25 kPA) of the collected water samples through pre-combusted (3-4 hours at 450 0C) Whatman GF/F glass fiber filters. The filtrate was stored in high-density polyethylene bottles and frozen (-20 0C) until analysis. Two replicates were run from the same sample bottle. Concentrations were determined using a Lachat Quick Chem 8000 flow injection autoanalyzer (Milwaukee, WI, USA). The method detection limit from the 10/6/99 through 10/21/02 sampling dates was 0.18 µM. The method detection limit beginning on the 11/21/02 sampling date was 1.24 µM. The mdl starting on sampling date of 10/24/2014 was 0.09 uM. Mdl on sampling date of 2/4/2015 was changed to 0.08 uM. MDL on sampling date of 8/14/2017 was changed to 0.03 uM. MDL of 1.17 on 1/25/2020 Parameter or Variable: CHLOROPHYLL A CONCENTRATION (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: fluorometer; Sampling and Analyzing Method: Phytoplankton chlorophyll a (chl a) concentrations were measured using the modified in vitro fluorescent technique in EPA Method 445.0 (Arar et al. 1997). Samples were collected on 25 mm Whatman GF/F glass fiber filters using vacuum filtration (< 25 kPa), blotted dry, and frozen immediately (-20 0C). Chlorophyll a was extracted from the filter using a tissue grinder and 90% aqueous acetone. The samples remained in the acetone overnight at -20 0C. The extracts were filter-clarified and analyzed on a TD700 fluorometer. The fluorometer was calibrated with chl a after determining the concentration using a Shimadzu UV160U Spectrophotometer and the extinction coefficients of Jeffrey and Humphrey (1975). The calibration was checked daily against a solid secondary standard (Turner Designs, proprietary formula). References: Arar, E.J., W.L. Budde, and T.D. Behymer. 1997. Methods for the determination of chemical substances in marine and environmental matrices. EPA/600/R-97/072. National Exposure Research Laboratory, U.S. Environmental Protection Agency, Cincinnati, Ohio. Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Chlorophyll a (Chl a) measurements prior to the 08/17/1999 sampling date were measured on a Shimadzu UV-160U spectrophotometer using the trichromatic equation following sonication (45-60 s) and overnight extraction of glass fiber filters in 90 % acetone. Beginning on the 08/17/1999 sampling date, Chl a concentration was measured using the modified in vitro fluorescence technique in EPA Method 445.0 (Welshmeyer 1994, Arar et al. 1997): Fifty milliliters of each water sample was vacuum filtered (less than 25 kilopascals) in duplicate at low ambient light conditions using 25 mm Whatman glass microfibre filters (GF/F). The filters were blotted dry, wrapped in foil and frozen immediately at -20 degrees Celsius until analysis. Chlorophyll a was extracted from the filter using a tissue grinder and 10 mL of 90 percent reagent grade aqueous acetone (v/v with deionized water, Fisher Scientific NF/FCC Grade). The samples remained in the acetone overnight at -20 degrees Celsius. The extracts were filter-clarified using a centrifuge and analyzed on a Turner Designs TD-700 fluorometer that was configured for the non-acidification method of Welschmeyer (1994) (EX 436/ EM 685) blue mercury lamp. The value reported is the average chlorophyll a concentration measured from the two filters. The fluorometer was calibrated with a known concentration of pure Chl a that was determined using a Shimadzu UV-160U spectrophotometer and the extinction coefficients of Jeffrey and Humphrey (1975). The calibration was checked daily against a solid secondary standard (Turner Designs, proprietary formula). As of August 2010, fluorescence was also measured on a TurnerDesigns Trilogy fluorometer. MDL 0.025 ug/l. 7200-046 Chl a extraction non acid module, 436 EX/685 EM (460 LED). References: 1. Welschmeyer, N.A. 1994. Fluorometric analysis of chlorophyll a in the presence of chlorophyll b and pheopigments. Limnol. Oceanogr. 39:1985-1992. 2. Arar, E.J., W.L. Budde, and T.D. Behymer. 1997. Methods for the determination of chemical substances in marine and environmental matrices. EPA/600/R-97/072. National Exposure Research Laboratory, U.S. Environmental Protection Agency, Cincinnati, Ohio. 3. Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Parameter or Variable: assimilation index (calculated); Units: milligrams of carbon per milligrams of chlorophyll a per hour; Observation Category: other; Sampling Instrument: none; Sampling and Analyzing Method: Primary productivity divided by chlorophyll a (measured by in vitro fluorometric method). Parameter or Variable: Chlorophyllide a (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: bottle; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's reponse factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton R Parameter or Variable: YSI Time (measured); Units: Exact time (hours:minutes:seconds); Observation Category: in situ; Sampling Instrument: YSI - handheld multi-parameter instrument Parameter or Variable: uncorrected Chromophoric Dissolved Organic Matter (CDOM) (measured); Units: microgram/liter; Observation Category: in situ; Sampling Instrument: fluorometer; Sampling and Analyzing Method: Colored dissolved organic matter (CDOM) was measured using a Turner Designs TD-700 fluorometer configured with a near-UV mercury vapour lamp, a 350 nm excitation filter, and a 410–600 nm emission filter. The fluorometer was calibrated to quinine sulfate (QS) solutions made up in 2 N sulfuric acid. Water samples were vacuum filtered (less than 25 kilopascal) using pre-combusted Whatman glass microfibre filters (GF/F) and the filtrate was stored in scintillation vials in the dark at 4 degrees Celsius until fluorometric analysis. The official decision (3/2/2017) is that cdom results from 12/1/2003 through 4/25/2011 would be multiplied by a corrective factor of 2.0. Results for sample date of 5/9/2011 and after do not need correcting. It is believed the stock solution was made wrong, making a 1L recipe for 600 ug/L in a 500 ml flask equals 1200 ug/L stock solution. Standards were still calibrated according to recipe, but were actually 2x as strong. Parameter or Variable: nitrogen to phosphorus molar ratio (calculated); Units: ratio; Observation Category: other; Sampling Instrument: na; Sampling and Analyzing Method: The molar ratio of nitrogen (N) to phosphorus (P), or N:P, was calculated by dividing dissolved inorganic nitrogen (DIN) by orthophosphate (PO43-) concentrations. Parameter or Variable: CHLOROPHYLL A - INTEGRATED (calculated); Units: microgram/liter; Observation Category: in situ; Sampling Instrument: integrated water sampler; Sampling and Analyzing Method: Chlorophyll a concentration measured by in vitro fluorometry (micrograms per liter) integrated throughout the water column to 2x the secchi depth. Water samples for this measurement were collected using the integrated water sampler (IWS) which collects vertically integrated water samples. Parameter or Variable: PRIMARY PRODUCTIVITY (measured); Units: milligram C per meter cubed per hour; Observation Category: laboratory analysis; Sampling Instrument: field light simulator; Sampling and Analyzing Method: Primary Productivity rate was measured using an adaptation of Steeman Nielsen's (1952) 14C bicarbonate method (Paerl et al. 1998). This method of measuring primary productivity allows direct measurement of carbon uptake and measures only net photosynthesis: Water samples were stored in 10 Liter high density polyethylene containers overnight in the research pond, a flow through system that receives water from the adjacent Bogue Sound, thereby simulating ambient water temperatures. The following morning the water samples were removed from the pond and transported to the laboratory for analysis. Water samples (76 milliliters) were added to three clear plastic square bottles to determine light uptake of carbon in triplicate and to 1 dark bottle to determine dark uptake of carbon. A solution of radioactive carbonate (300 microliters) was added to each bottle. The bottles were incubated for 4 hours in the pond. The light bottles were incubated underneath a field light simulator, while the dark bottles were incubated in a covered perforated bucket that was submerged in the pond. The FLS was used to simulate the ambient light conditions that phytoplankton are exposed to in the estuary (mixing conditions). The FLS is comprised of a rotating wheel with varying levels of screening. During the incubation period, photosynthetically active radiation (PAR) measurements were performed using a 2 pi Li-Cor LI-192SA spherical quantum sensor attached to a Li-Cor data logger. After the incubation period, the samples were returned to the laboratory, shaken and the entire contents were gently vacuum filtered (less than 25 kilopascals) using 25 mm Whatman glass microfibre filters (GF/F). The filters were placed in wooden drying trays and treated with concentrated hydrochloric acid fumes for 40 minutes to an hour to remove inorganic 14C. The filters were folded in half and placed in 7 milliliter plastic scintillation vials. Five milliliters of liquid scintillation cocktail (ecolume or cytoscint) was added to the vials. The vials were capped, shaken, stored in the dark for 3-24 hours and then assayed for radioactivity using a Beckman liquid scintillation counter. In addition to the samples, triplicate voucher samples were used to quantify the radioactivity of the 14C added. Voucher samples consisted of 100 microliter of 14C and 100 microliters of phenylethylamine. These vials also received 5 milliliters of liquid scintillation cocktail. A background vial and two 14C background standards were used. The quantity of carbon fixed is proportional to the fraction of radioactive carbon assimilated. (Paerl, H.W., J.L. Pinckney, J.M. Fear, and B.L. Peierls 1998. Ecosystem responses to internal and watershed organic matter loading: consequences for hypoxia in the eutrophying Neuse River Estuary, North Carolina, USA. Marine Ecology Progress Series 166: 17-25; Steemann Nielsen, E. 1952. The use of radio-active carbon (C14) for measuring organic production in the sea. Journal du Conseil permanent international pour L'Exploration de la Mer 18: 117-140) Parameter or Variable: Chlorophyllide a (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: chlorophyll c1 and c2 (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Peridinin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: 19'-Butanoyloxyfucoxanthin (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Pheophorbide a (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: fucoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: 19'-Hexanoyloxyfucoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: 9'-cis Neoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Prasinoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Violaxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Diadinoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: antherexanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Myxoxanthophyll (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: alloxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: diatoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: monadoxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Lutein (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Zeaxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: gyroxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: canthaxanthin (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: CHLOROPHYLL A CONCENTRATION (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: divinyl chlorophyll b (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: chlorophyll a corrected (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: echinenone (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: pheophytin a (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: beta-carotene (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: Total CHLOROPHYLL A (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: Diagnostic phytoplankton photopigments were identified, separated and quantified by high performance liquid chromatography coupled to an in-line photodiode array spectrophotometer (Jeffrey et al. 1997): Known volumes of water sample (500-1000 milliliters, enough to obtain color on the filter) were vacuum filtered (less than 25 kiloPascals) through 25 or 47 millimeter Whatman glass microfibre filters (GF/F) under reduced light conditions. The filters were blotted dry, folded in half, wrapped in foil and then immediately frozen at -20 degrees Celsius until analysis. The filters were placed in 15 milliliter centrifuge tubes containing 1.5-3.0 milliliters of 100% acetone (HPLC Grade), sonicated for 30-60 seconds using a Fisher Sonic Dismembrator 300 with microtip and extracted at -20 degrees Celsius for 12-24 hours. After extraction the samples were centrifuged at 4500 rpm and the supernatant (i.e.- the combined extracted pigments) collected & filtered into amber glass autosampler vials using Millipex Millipore 0.45 micometer PTFE. Two hundred microliters of extractant from each vial was injected into the HPLC system using a Spectra Physics (now Thermo Separations Products) AS3000 autosampler and SP8800 pump, running a non-linear, 55 minute, 2-solvent gradient adapted from Van Heukelem et.al. 1994 or 1995?. The nonlinear, variable flow, binary gradient consisted of solvent A [80% methanol : 20% ammonium acetate (0.5 M adjusted to pH 7.2)] and B (80% methanol : 20% acetone). The extractant was separated into individual pigments using a series of C18 reverse-phase columns to optimize photopigment separations: The column order was a Rainin Microsorb guard column (0.46 x 1.5 centimeters, 3 micrometer packing) followed by a single monomeric reverse-phase C18 column (Rainin Microsorb-MV, 0.46 x 10 cm, 3 µm packing) followed by two polymeric reverse-phase C18 columns (Vydac 201TP5, 0.46 x 25 cm, 5 µm packing). The columns were kept at a constant 52 degrees Celsius in an Alltech 330 column heater. The separated pigments were then passed through an in line Shimadzu SPD-M10AV photodiode array detector which measured the absorbance of the sample/extractant, scanning the range of 350-800 nanometers every 2 seconds. The data was collected and analyzed using Shimadzu's EZChrom software. Individual pigments are identified using a combination of peak retention time and absorbance spectrum shape. Retention times and absorbance spectra are identified for each pigment by analyzing known pigments (either as pure standards or pigments or isolated from algal cultures). Pigments are quantified from their peak areas, calculated at 440nm. A calibration curve is generated by injecting various volumes of a mixed standard composed of known quantities of seven pure pigment standards (fucoxanthin, zeaxanthin, bacteriochlorophyll a, canthaxathin, chlorophyll b, chlorophyll a, echinenone and ß-carotene) and calculating the peak areas of those pigments The peak areas are regressed against the known quantities of each pigment to calculate the slope (Response Factor) for that pigment. Response factors for pigments we do not have reference standards for are calculated using the ratio of absorbance coefficients of each pigment to its closest structurally related reference pigment, multiplying the known pigment's response factor by that ratio. Pigments extracted from the samples are then quantified by multiplying the peak areas of a chromatogram at 440nm by the response factors. Pigment values listed as below detection were below the software threshold for peak detection or had spectra below a similarity of 0.9 compared to library spectra. Technician expert judgement was used in difficult cases. References: Jeffrey, S.W., R.F.C. Mantoura, and S.W. Wright. 1997. Phytoplankton pigments in oceanography: Guidelines to modern methods. UNESCO Publishing, Paris, France. Pinckney, J.L., D.F. Millie, K.E. Howe, H.W. Paerl, and J. P. Hurley. 1996. Flow scintillation counting of 14C-labeled microalgal photosynthetic pigments. Journal of Plankton Research 18:1867-1880. Parameter or Variable: chlorophytes (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: The HPLC derived diagnostic photopigment concentrations were analyzed using the ChemTax matrix factorization program (Mackey 1996). This program uses the steepest decent algorithm to determine the best fit based on an initial estimate of pigment ratios for algal classes. The initial pigment ratio matrix used in the Chemtax analysis was derived from: Mackey M.D., Mackey D.J., Higgins H.W., & Wright S.W. 1996. CHEMTAX- a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265-283, and consisted of nine photopigments (alloxanthin, antheraxanthin, chlorophyll b, total chlorophyll a (chlorophyll a + chlorophyllide a), fucoxanthin, lutein, peridinin, violaxanthin, and zeaxanthin) for five algal groups that constitute the bulk of the phytoplankton community in the Neuse River and Estuary (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates). In order to reduce the variation of pigment ratios due to large changes in phytoplankton species composition with depth, season, and salinity regime, homogenous data groupings of the HPLC pigment data were performed prior to running on Chemtax: HPLC pigment data was grouped by Depth Level (surface or bottom) then by Season (winter, spring, summer and fall) then by Salinity regime (oligohaline: <5.0 ppt, mesohaline: 5.01 - 18.0 ppt, polyhaline: >18.01 ppt). When there were less than 10 samples in a given homogenous grouping (Chemtax requires at least 10 samples per run), the data was grouped by oligohaline + mesohaline or mesohaline + polyhaline (This is indicated in the comments section). Parameter or Variable: Cryptophytes (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: The HPLC derived diagnostic photopigment concentrations were analyzed using the ChemTax matrix factorization program (Mackey 1996). This program uses the steepest decent algorithm to determine the best fit based on an initial estimate of pigment ratios for algal classes. The initial pigment ratio matrix used in the Chemtax analysis was derived from: Mackey M.D., Mackey D.J., Higgins H.W., & Wright S.W. 1996. CHEMTAX- a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265-283, and consisted of nine photopigments (alloxanthin, antheraxanthin, chlorophyll b, total chlorophyll a (chlorophyll a + chlorophyllide a), fucoxanthin, lutein, peridinin, violaxanthin, and zeaxanthin) for five algal groups that constitute the bulk of the phytoplankton community in the Neuse River and Estuary (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates). In order to reduce the variation of pigment ratios due to large changes in phytoplankton species composition with depth, season, and salinity regime, homogenous data groupings of the HPLC pigment data were performed prior to running on Chemtax: HPLC pigment data was grouped by Depth Level (surface or bottom) then by Season (winter, spring, summer and fall) then by Salinity regime (oligohaline: <5.0 ppt, mesohaline: 5.01 - 18.0 ppt, polyhaline: >18.01 ppt). When there were less than 10 samples in a given homogenous grouping (Chemtax requires at least 10 samples per run), the data was grouped by oligohaline + mesohaline or mesohaline + polyhaline (This is indicated in the comments section). Parameter or Variable: cyanobacteria (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: The HPLC derived diagnostic photopigment concentrations were analyzed using the ChemTax matrix factorization program (Mackey 1996). This program uses the steepest decent algorithm to determine the best fit based on an initial estimate of pigment ratios for algal classes. The initial pigment ratio matrix used in the Chemtax analysis was derived from: Mackey M.D., Mackey D.J., Higgins H.W., & Wright S.W. 1996. CHEMTAX- a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265-283, and consisted of nine photopigments (alloxanthin, antheraxanthin, chlorophyll b, total chlorophyll a (chlorophyll a + chlorophyllide a), fucoxanthin, lutein, peridinin, violaxanthin, and zeaxanthin) for five algal groups that constitute the bulk of the phytoplankton community in the Neuse River and Estuary (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates). In order to reduce the variation of pigment ratios due to large changes in phytoplankton species composition with depth, season, and salinity regime, homogenous data groupings of the HPLC pigment data were performed prior to running on Chemtax: HPLC pigment data was grouped by Depth Level (surface or bottom) then by Season (winter, spring, summer and fall) then by Salinity regime (oligohaline: <5.0 ppt, mesohaline: 5.01 - 18.0 ppt, polyhaline: >18.01 ppt). When there were less than 10 samples in a given homogenous grouping (Chemtax requires at least 10 samples per run), the data was grouped by oligohaline + mesohaline or mesohaline + polyhaline (This is indicated in the comments section). Parameter or Variable: diatom (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: The HPLC derived diagnostic photopigment concentrations were analyzed using the ChemTax matrix factorization program (Mackey 1996). This program uses the steepest decent algorithm to determine the best fit based on an initial estimate of pigment ratios for algal classes. The initial pigment ratio matrix used in the Chemtax analysis was derived from: Mackey M.D., Mackey D.J., Higgins H.W., & Wright S.W. 1996. CHEMTAX- a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265-283, and consisted of nine photopigments (alloxanthin, antheraxanthin, chlorophyll b, total chlorophyll a (chlorophyll a + chlorophyllide a), fucoxanthin, lutein, peridinin, violaxanthin, and zeaxanthin) for five algal groups that constitute the bulk of the phytoplankton community in the Neuse River and Estuary (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates). In order to reduce the variation of pigment ratios due to large changes in phytoplankton species composition with depth, season, and salinity regime, homogenous data groupings of the HPLC pigment data were performed prior to running on Chemtax: HPLC pigment data was grouped by Depth Level (surface or bottom) then by Season (winter, spring, summer and fall) then by Salinity regime (oligohaline: <5.0 ppt, mesohaline: 5.01 - 18.0 ppt, polyhaline: >18.01 ppt). When there were less than 10 samples in a given homogenous grouping (Chemtax requires at least 10 samples per run), the data was grouped by oligohaline + mesohaline or mesohaline + polyhaline (This is indicated in the comments section). Parameter or Variable: Dinoflagellates (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: The HPLC derived diagnostic photopigment concentrations were analyzed using the ChemTax matrix factorization program (Mackey 1996). This program uses the steepest decent algorithm to determine the best fit based on an initial estimate of pigment ratios for algal classes. The initial pigment ratio matrix used in the Chemtax analysis was derived from: Mackey M.D., Mackey D.J., Higgins H.W., & Wright S.W. 1996. CHEMTAX- a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265-283, and consisted of nine photopigments (alloxanthin, antheraxanthin, chlorophyll b, total chlorophyll a (chlorophyll a + chlorophyllide a), fucoxanthin, lutein, peridinin, violaxanthin, and zeaxanthin) for five algal groups that constitute the bulk of the phytoplankton community in the Neuse River and Estuary (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates). In order to reduce the variation of pigment ratios due to large changes in phytoplankton species composition with depth, season, and salinity regime, homogenous data groupings of the HPLC pigment data were performed prior to running on Chemtax: HPLC pigment data was grouped by Depth Level (surface or bottom) then by Season (winter, spring, summer and fall) then by Salinity regime (oligohaline: <5.0 ppt, mesohaline: 5.01 - 18.0 ppt, polyhaline: >18.01 ppt). When there were less than 10 samples in a given homogenous grouping (Chemtax requires at least 10 samples per run), the data was grouped by oligohaline + mesohaline or mesohaline + polyhaline (This is indicated in the comments section). Parameter or Variable: Total Chlorophyll a (calculated); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: not provided; Sampling and Analyzing Method: The HPLC derived diagnostic photopigment concentrations were analyzed using the ChemTax matrix factorization program (Mackey 1996). This program uses the steepest decent algorithm to determine the best fit based on an initial estimate of pigment ratios for algal classes. The initial pigment ratio matrix used in the Chemtax analysis was derived from: Mackey M.D., Mackey D.J., Higgins H.W., & Wright S.W. 1996. CHEMTAX- a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265-283, and consisted of nine photopigments (alloxanthin, antheraxanthin, chlorophyll b, total chlorophyll a (chlorophyll a + chlorophyllide a), fucoxanthin, lutein, peridinin, violaxanthin, and zeaxanthin) for five algal groups that constitute the bulk of the phytoplankton community in the Neuse River and Estuary (chlorophytes, cryptophytes, cyanobacteria, diatoms, and dinoflagellates). In order to reduce the variation of pigment ratios due to large changes in phytoplankton species composition with depth, season, and salinity regime, homogenous data groupings of the HPLC pigment data were performed prior to running on Chemtax: HPLC pigment data was grouped by Depth Level (surface or bottom) then by Season (winter, spring, summer and fall) then by Salinity regime (oligohaline: <5.0 ppt, mesohaline: 5.01 - 18.0 ppt, polyhaline: >18.01 ppt). When there were less than 10 samples in a given homogenous grouping (Chemtax requires at least 10 samples per run), the data was grouped by oligohaline + mesohaline or mesohaline + polyhaline (This is indicated in the comments section). Parameter or Variable: Corrected Chromophoric Dissolved Organic Matter (CDOM) (measured); Units: microgram/liter; Observation Category: laboratory analysis; Sampling Instrument: fluorometer; Sampling and Analyzing Method: The official decision (3/2/2017) is that cdom results from 12/1/2003 through 4/25/2011 would be multiplied by a corrective factor of 2.0. Results for sample date of 5/9/2011 and after do not need correcting. It is believed the stock solution was made wrong, making a 1L recipe for 600 ug/L in a 500 ml flask equals 1200 ug/L stock solution. Standards were still calibrated according to recipe, but were actually 2x as strong. Parameter or Variable: PAR Depth (measured); Units: meter; Observation Category: in situ; Sampling Instrument: UWO-PAR 6067; Sampling and Analyzing Method: Prior to the 09/13/2000 sampling date, in situ measurements were performed at discrete depths at approximately 0.5 meter increments using a Hydrolab Data Sonde 3 equipped with a multiprobe and SVR3 display logger. Beginning on the 09/13/2000 sampling date, in situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, 6600 EDS-S Extended Deployment System, 6600 V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561 or 6566), a YSI pulsed dissolved oxygen probe (Model 6562), a self cleaning YSI turbidity probe (Model 6026 or 6136), and beginning on the 07/30/2003 sampling date, a flat Li-Cor sensor (UWQ-PAR 6067). The YSI sonde was coupled to a either a YSI 610 DM datalogger or a YSI 650 MDS Multi-parameter Display System datalogger. In situ measurements from near surface (approximately 0.2 meters) and near bottom of the water column (approximately 0.5 meters from the sediment layer) are extracted to be included with the water sample data in "NRWQ_WorkingUpdate.xlsx". The YSI data were stored on the datalogger and downloaded to a PC using YSI EcoWatch software upon return to the laboratory. Parameter or Variable: PHOTOSYNTHETIC ACTIVE RADIATION (PAR) - Channel 1 (measured); Units: watt per meter; Observation Category: in situ; Sampling Instrument: UWO-PAR 6067; Sampling and Analyzing Method: Prior to the 09/13/2000 sampling date, in situ measurements were performed at discrete depths at approximately 0.5 meter increments using a Hydrolab Data Sonde 3 equipped with a multiprobe and SVR3 display logger. Beginning on the 09/13/2000 sampling date, in situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, 6600 EDS-S Extended Deployment System, 6600 V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561 or 6566), a YSI pulsed dissolved oxygen probe (Model 6562), a self cleaning YSI turbidity probe (Model 6026 or 6136), and beginning on the 07/30/2003 sampling date, a flat Li-Cor sensor (UWQ-PAR 6067). The YSI sonde was coupled to a either a YSI 610 DM datalogger or a YSI 650 MDS Multi-parameter Display System datalogger. In situ measurements from near surface (approximately 0.2 meters) and near bottom of the water column (approximately 0.5 meters from the sediment layer) are extracted to be included with the water sample data in "NRWQ_WorkingUpdate.xlsx". The YSI data were stored on the datalogger and downloaded to a PC using YSI EcoWatch software upon return to the laboratory. Parameter or Variable: PHOTOSYNTHETIC ACTIVE RADIATION (PAR) - Channel 2 (measured); Units: watt per meter; Observation Category: in situ; Sampling Instrument: UWO-PAR 6067; Sampling and Analyzing Method: Prior to the 09/13/2000 sampling date, in situ measurements were performed at discrete depths at approximately 0.5 meter increments using a Hydrolab Data Sonde 3 equipped with a multiprobe and SVR3 display logger. Beginning on the 09/13/2000 sampling date, in situ measurements were performed at discrete depths on the sunlit side of the research vessel using a Yellow Springs Instruments (YSI Incoporated, Ohio) multiparameter sonde (Model 6600, 6600 EDS-S Extended Deployment System, 6600 V2-4) equipped with a YSI conductivity/temperature probe (Model 6560), a YSI chlorophyll probe (Model 6025), a YSI pH probe (Model 6561 or 6566), a YSI pulsed dissolved oxygen probe (Model 6562), a self cleaning YSI turbidity probe (Model 6026 or 6136), and beginning on the 07/30/2003 sampling date, a flat Li-Cor sensor (UWQ-PAR 6067). The YSI sonde was coupled to a either a YSI 610 DM datalogger or a YSI 650 MDS Multi-parameter Display System datalogger. In situ measurements from near surface (approximately 0.2 meters) and near bottom of the water column (approximately 0.5 meters from the sediment layer) are extracted to be included with the water sample data in "NRWQ_WorkingUpdate.xlsx". The YSI data were stored on the datalogger and downloaded to a PC using YSI EcoWatch software upon return to the laboratory. asNeeded Metadata are developed, maintained and distributed by NCEI. Updates are performed as needed to maintain currentness. NOAA National Centers for Environmental Information custodian bottle bottle any type of water bottle sampling device Generic name for water collection device; usually used to determine temperature, salinity and provide water aliquots for measurement of a wide range of parameters; often referred to by a specific type of water sampling bottle, such as a Nansen or Niskin bottle. flow injection analyzer flow injection analyzer automated chemical analysis instrument An instrument that performs flow injection analysis. Flow injection analysis (FIA) is an approach to chemical analysis that is accomplished by injecting a plug of sample into a flowing carrier stream. FIA is an automated method in which a sample is injected into a continuous flow of a carrier solution that mixes with other continuously flowing solutions before reaching a detector. Precision is dramatically increased when FIA is used instead of manual injections and as a result very specific FIA systems have been developed for a wide array of analytical techniques. https://www.bco-dmo.org/instrument/657 (accessed 2018-01-26) Laboratory instrument that analyzes multiple media for ions. The media include various solids and liquids. Models include: QuikChem 8000 Series FIA System fluorometer fluorometer fluorometer oxygen sensor oxygen sensor Any instrument that measures oxygen content in a compound. PAR Sensor PAR Sensor Photosynthetically Active Radiation Sensor The PAR sensor measures global solar radiation from 400 to 700 nm, which approximates the spectral band active in photosynthesis. The response of the instrument falls sharply to zero on either side of this band, and between 400 and 670 nm it increases monotonically from about 50% to 100%. There is no protective glass dome over the receiving surface. Irradiance from the sun passes through a small white visible bandpass filter/diffuser in the shape of a horizontal disk at the top of the instrument. Within the instrument radiation is directed through a series of colored glass filters and onto a silicon photodiode detector. The factory-supplied calibration converts the signal from the detector to a flux of photons in micro-mole (of photons) s-1 m-2. (One micro-mole equals 6.022 E17 photons.) For the solar spectrum, these units may be converted to watts per square meter by dividing by 4.6. (http://www.srrb.noaa.gov/instrument/par.htm) pH sensor pH sensor generic meter used to measure pH Any of a class of instruments used to quantify pH in a water or other type of sample. Secchi disk Secchi disk The Secchi disk (or Secchi disc), as created in 1865 by Angelo Secchi, is a plain white, circular disk 30 cm (12 in) in diameter used to measure water transparency or turbidity in bodies of water. The disc is mounted on a pole or line, and lowered slowly down in the water. The depth at which the disk is no longer visible is taken as a measure of the transparency of the water. This measure is known as the Secchi depth and is related to water turbidity. [Wikipedia, 2021-09-02] thermometer thermometer generic thermometer or reversing thermometer use thermometer when temperature is a data type, but the exact instrument used to measure temperature is unknown Total Organic Carbon (TOC) analyzer Total Organic Carbon (TOC) analyzer measures the total amount of organic carbon contained in water turbidity sensor turbidity sensor Measures the turbidity of water Turbidity sensors measure the amount of light that is scattered by suspended solids in a liquid, such as water. When the concentration of total suspended solids (TSS) and total dissolved solids (TDS) in a liquid increase, the turbidity also increases.