hplc pigment processing update crystal thomas science systems and applications, inc. (ssai)...
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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 farRunning sets in the order in which they were
collectedNASA PIs first
Data ReportingAnalysis precision
Duplicate filter precision
Various sums, ratios, and indices
23 pigments, Total chlorophyll a, b and cACE 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
on
se (
mA
u)
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
Many thanks to my group:
Joaquin Chaves Chuck McClain Aimee NeeleyGiulietta FargionAntonio Mannino Mike NovakScott Freeman Karen Mitchell Chris ProctorChris Kenemer