HPLC Pigment Processing Update - Ocean color

HPLC Pigment Processing Update 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 -80C freezer with LN backup and emergency call system 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 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 Sample Status 3760 samples reported by PIs 1180 analyzed so far Running sets in the order in which they were collected NASA PIs first

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 Developing new HPLC analysis method

Response (mAu) Current method New method Retention time (min) 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 Want more information? http://oceancolor.gsfc.nasa.gov/HPLC/

Information on shipping samples, reporting practices, pigment list Website is a work in progress [email protected] 301-286-7299 Many thanks to my group: Joaquin Chaves Giulietta Fargion Scott Freeman Chris Kenemer Chuck McClain Antonio Mannino Karen Mitchell

Aimee Neeley Mike Novak Chris Proctor

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