ReadMe file: SBI: HLY-02-01 and HLY-02-03 data from Dr. Glenn Cota and Dr. Victoria Hill TITLE: HPLC pigment data AUTHORS: Dr Glenn F. Cota Center for Coastal Physical Oceanography Old Dominion University Norfolk VA 23529 Dr. Victoria J. Hill Department of Ocean Earth and Atmospheric Science, ODU 4600 Elkhorn Ave Norfolk VA 23529 757.683.4911 (tel) 757.683.5303 (fax) email vhill@odu.edu Data questions contact: Dr V. J. Hill FUNDING SOURCE AND GRANT NUMBER: NSF – OPP – 0125049 DATA SET OVERVIEW: High Performance Liquid Chromatography (HPLC) pigment data Data is from two cruises in the Chukchi and Beaufort Seas onboard the USCGC Healy WAGB-20 during the spring (HLY-02-01) and summer (HLY-02-03) of 2002 as part of the SBI phase II project. A summary list of Bio-Optical stations and locations from the two cruises are given in file Stn_List_Arc2002.txt. Detailed station information can be found in the individual station sheets. Station sheets and photos are listed by year, month, day, and order for that day. Time period covered by the data: Spring cruise (HLY-02-01) 8 May 2002 to 13 June 2002. Summer cruise (HLY-02-03) 18 July 2002 to 21 August 2002. Physical location of the measurement See Stn_List_Arc2002.txt for summary list of station locations. Spring cruise (HLY-02-01) Arc2002-1 Min Lat: 65deg 01.380min; Max Lat: 73deg 47.040min (North) Min Lon: -169deg 03.105min; Max Lon: -154deg 13.081min (West) Summer cruise (HLY-02-03) Arc2002-2 Min Lat: 67deg 30.262min; Max Lat: 73deg 40.439min (North) Min Lon: -168deg 54.675min; Max Lon: -151deg 46.506min (West) INSTRUMENT DESCRIPTION: Analysis of samples was performed by J. R. Perl and C. C. Trees at the Center for Hydro-Optics and Remote Sensing, San Diego State University. The HPLC method used was that proposed by Wright et al. (1991). Pigments were separated on the ODS- 2 C18 column using a three solvent gradient system at a flow rate of one ml per min. The separation of the various pigments requires 25 minutes with the pigment peaks being detected by a absorption detector; a ThermoQuest UV6000 scanning diode array detector (190 to 800 nm at 1 nm resolution). In addition, a ThermoQuest FL3000 scanning fluorescence detector was used to detect and quantify the various chlorophyll degradation products, which occur at lower concentrations. DATA COLLECTION and PROCESSING: Samples were collected at 6 depths throughout the water column in parallel with productivity samples, which is 100-80, 50, 30, 15, 5, and 1% of the surface irradiance. Samples were filtered in low light and temperature conditions onto 25mm GF/F filters (nominal pore size 0.7um), and placed immediately in the -80oC freezer. They were then shipped to San Diego in dry ice. They were extracted in 4 mls of 100% acetone. With a filter and sample the water retention is about 0.2 to 0.3 ml and equates to about a 92-95% acetone extraction solution. After 24 hours of extraction in a freezer (-20oC), the samples were sonicated for 10 seconds using a microprobe tip at a 60% duty cycle. They were then extracted again for 24 hours. Glass-fiber particles, generated during sonication, were removed from the extract by centrifugation and filtration using 0.2 um PTFE in-line filters. An internal pigment standard (canthaxanthin, which is normally not found in samples) was added to the extract to correct for any extraction volume changes during sample processing. Since canthaxanthin is a carotenoid and does not fluoresce, it does not affect the fluorometric analysis. The standard fluorometric method of Holm-Hansen et al. (1965) was used to calculate chlorophyll and phaeopigment concentrations on an aliquot (100 ul) of the pigment extract. These concentrations were also correct for extraction volume changes using the canthaxanthin internal standard. The NASA protocol for the Fluorometric Chlorophyll a: Sampling, Laboratory, Methods and Data Analysis (Trees et al., 2000) was followed. PIGMENT NOMENCLATURE (Updated 1/23/02 to match SeaBASS Field Names) Parameters Description Units Beta-epi-Car Alpha carotene mg m-3 Allo Alloxanthin mg m-3 Beta-beta-Car Beta carotene mg m-3 But-fuco 19’-Butanolyoxyfucoxanthin mg m-3 Chlide_a Chlorophylide a mg m-3 Chl_b Chlorophyll b mg m-3 Chl_c2 Chlorophyll c2 mg m-3 Chl_c1c2 Chlorophyll c1 + chlorophyll c2 mg m-3 Chl_c3 Chlorophyll c3 mg m-3 Diadino Diadinoxanthin mg m-3 Diato Diatoxanthin mg m-3 Fuco Fucoxanthin mg m-3 Hex-fuco 19’-Hexanoyloxyfucoxanthin mg m-3 Lut Lutein mg m-3 Perid Peridinin mg m-3 Pras Prasinoxanthin mg m-3 Viola Violaxanthin mg m-3 Zea Zeaxanthin mg m-3 Chl_a Chlorophyll a mg m-3 MV_Chl_a Monovinyl chlorophyll a mg m-3 DV_Chl_a Divinyl chlorophyll a mg m-3 Tot_Chl_a Monovinyl chlorophyll a mg m-3 + Divinyl chlorophyll a + Chlorophilide a + Chlorophyll a allomer + Chlorophyll a epimer DATA FORMAT: Data is in SeaBASS format (ASCII) tab delimited with a SeaBASS header. The headers include parameters, units, time, location, and other Meta data. DATA REMARKS: Missing or questionable data was not included in the submitted data set or is noted in the header information of the file. REFERENCES: Holm-Hansen, O., C.J. Lorenzen, R.W. Holmes and J.D.H. Strickland. 1965. Fluorometric determination of chlorophyll. J. Cons. Cons. Int. Explor. Mer., 30: 3-15. Latasa, M., R.R. Bidigare, M.E. Ondrusek and M.C. Kennicutt II. 1996. HPLC analysis of algal pigments: a comparison exercise among laboratories and recommendations for improved analytical performance. Mar Chem. 51: 315-324. Trees, C.C., R..R. Bidigare, D.N. Karl and L. Van Heukelem. 2000. Fluorometric chlorophyll a: sampling, laboratory methods, and data analysis protocols. In: G.S. Fargion and J.L. Mueller (Eds.) Ocean Optics Protocols for Satellite Ocean Color Sensor Validation, Revision 2, Chapter 14. NASA TM 2000-209966, Goddard Space Flight Center, Greenbelt, MD. pp:162-169. Wright, S.W., S.W. Jeffrey, R.F.C. Mantoura, C.A. Llewellyn, T. Bjornland, D. Repeta and N. Welschmeyer. 1991. Improved HPLC method for the analysis of chlorophylls and carotenoids from marine phytoplankton. Mar. Ecol. Prog. Ser. 77: 183.