HPLC P igment Processing U pdate

HPLC Pigment Processing Update

Crystal ThomasScience Systems and Applications, Inc. (SSAI)

NASA-Goddard Space Flight Center

23April2012NASA OCRT meeting

New labAgilent 1260 HPLC Agilent RR1200 HPLC

-80°C freezer with LN backup and

emergency call system

Turner Designs 10-AU

Re-validation of RR1200Calibration-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 HPLTo show results produced at GSFC will be

consistent with previous data setsSplit several sets of sample extracts as well as

standardsCalibrations 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 Status3760 samples reported by PIs 1180 analyzed so far

Running sets in the order in which they were collected

NASA PIs first

Data ReportingAnalysis precisionDuplicate filter precisionVarious sums, ratios, and indices23 pigments, Total chlorophyll a, b and c

ACE working group has discussed potential changes to reported pigment listSuggest removing prasinoxanthin, lutein, gyroxanthin

diester from routine quantitation

Developing new HPLC analysis method

Current method

New methodResp

onse

(m

Au)

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 listWebsite is a work in progress

crystal.s.thomas@nasa.gov301-286-7299

Many thanks to my group:

Joaquin Chaves Chuck McClain Aimee NeeleyGiulietta FargionAntonio Mannino Mike NovakScott Freeman Karen Mitchell Chris ProctorChris Kenemer