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HPLC P igment Processing U pdate

HPLC P igment Processing U pdate. Crystal Thomas Science Systems and Applications, Inc. (SSAI) NASA-Goddard Space Flight Center. 23April2012 NASA OCRT meeting. New lab. Agilent 1260 HPLC. Agilent RR1200 HPLC. Turner Designs 10-AU. -80°C freezer with LN backup and

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HPLC P igment Processing U pdate

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  1. HPLC Pigment Processing Update Crystal Thomas Science Systems and Applications, Inc. (SSAI) NASA-Goddard Space Flight Center 23April2012 NASA OCRT meeting

  2. New lab Agilent 1260 HPLC Agilent RR1200 HPLC Turner Designs 10-AU -80°C freezer with LN backup and emergency call system

  3. Re-validation of RR1200 • Calibration-single point and multi-point (when possible) • Dilution series of standards and samples to test linearity, range • Comparison of QC data with performance metric levels established in SeaHARRE – overall rating “state-of-the-art” • Quantitative – Rs, RT precision, solvent delivery precision • State-of-the-art – Total Chl a and primary pigment precision and accuracy, injection precision, calibration accuracy • Comparison of calibration with HPL

  4. Comparison with HPL • To show results produced at GSFC will be consistent with previous data sets • Split several sets of sample extracts as well as standards • Calibrations of all quantitated pigments agree within 6.5% (85% within 5%) • Total Chl a agreement = 0.2% • SeaHARRE “state-of-the-art” levels • Within 10% for primary pigments • Within 2% for Total Chl a

  5. Sample Status • 3760 samples reported by PIs • 1180 analyzed so far • Running sets in the order in which they were collected • NASA PIs first

  6. Data Reporting • Analysis precision • Duplicate filter precision • Various sums, ratios, and indices • 23 pigments, Total chlorophyll a, b and c • ACE working group has discussed potential changes to reported pigment list • Suggest removing prasinoxanthin, lutein, gyroxanthin diester from routine quantitation

  7. Developing new HPLC analysis method Current method Response (mAu) New method Retention time (min)

  8. Still to be done on the new method • Decide if separation should be improved further • Injector program • Full calibration • Validation • Ruggedness, transferability • Linearity • Precision • Accuracy • Split extracts between 1200 and 1260 • Split extracts between GSFC and HPL • Replicate filters between GSFC and HPL • Use in SeaHARRE-6

  9. Want more information? http://oceancolor.gsfc.nasa.gov/HPLC/ Information on shipping samples, reporting practices, pigment list Website is a work in progress crystal.s.thomas@nasa.gov 301-286-7299 Many thanks to my group: Joaquin Chaves Chuck McClain Aimee Neeley Giulietta Fargion Antonio Mannino Mike Novak Scott Freeman Karen Mitchell Chris Proctor Chris Kenemer

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