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 HPLCAgilent RR1200 HPLC -80°C freezer with LN backup and emergency call system Turner Designs 10-AU

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 Current method New method Response (mAu) 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? Information on shipping samples, reporting practices, pigment list Website is a work in progress Many thanks to my group: Joaquin ChavesChuck McClainAimee Neeley Giulietta FargionAntonio ManninoMike Novak Scott FreemanKaren MitchellChris Proctor Chris Kenemer