MODIS Science Team Meeting

1
HPLC Phytoplankton Pigment Measurements
Continuation of Climate Quality Data Records
(CQDR) in Support of NASAs Earth Science
Enterprise (ESE) Chuck Trees and Jason
Perl Center for Hydro-Optics and Remote Sensing
(CHORS) San Diego State University (ctrees_at_chors.s
dsu.edu) (jperl_at_chors.sdsu.edu)
2
HPLC Phytoplankton Pigment Protocols (Bidigare et
al., 2003 NASA/TM-2003-211621/Rev 4 - Vol. V)

• EOS Terra Aqua Data Products Program - 3,000
  samples/yr
• Augmentation for an additional 1,800/yr
  (4,800/yr)
• 1. Van Heukelem and Thomas (2001)
• Filtration
• 1. GF/F glass fiber filters (25 mm)
• 2. Positive pressure filtration
• 3. Filtration Volumes
• a. Oligotrophic waters - 3-4 liters
• b. Mesotrophic waters - 1-2 liters
• c. Eutrophic waters - 0.5-1 liter

3
HPLC Phytoplankton Pigment Protocols

• Sample Handling, Storage and Shipping
• 1. Fold filters in half with filtered sides
  facing in.
• 2. Place in aluminum foil envelops, cryogenic
  vials or
• HistoPrep tissue capsules.
• 3. Label samples containers with a sequential
  number
• 4. Place and ship (dry shipper) in liquid
  nitrogen (LN2).
• Recordkeeping (electronically transmitted to
  CHORS).
• 1. For each sample record volume filtered,
  station
• number, date, time, depth, lat
  and long.

4
Template for Ancillary Data
5
HPLC System Calibrations
145 pigment compound calibration curves with
6-10 injections per curve (870-1,450 std
injections or 24-40 days of continuous running
standards).
6
Fluorometer Calibration
7
Number of Samples by Date and PI.
8
Number of Samples Analyzed by PI
9
Number of Samples Pending
10
Vitamin E Internal Standard (284 nm)
trans-?-apo-8'-carotenal (Fluka Chemical
Company)
Problems - Vitamin E Internal Standard
Vitamin E
Monovinyl Chl a
Divinyl Chl a
Divinyl Chl b
Zeaxanthin
b,b Carotene
11
Mixing the sample with the buffer. Heukelem
and Thomas (2001) method picks up in a train
(150-75-75-75-150). It is mixed when injected
onto the column. CHORS HPLC System mixes in
a 2nd vial. Minimized the mixing time, but the
uncertainty increased 3-4. 2. UV/VIS halogen
lamp. Faulty connection at the base of the lamp
which would cause erratic readings. Solution was
to have several lamps in stock to test when the
lamp failed.
Other Problems with HPLC Method and System
12
Culture Mixture (450 nm)
Mixture of Pigments from Cultures
Fucoxanthin
Monovinyl Chl a
Chl c3
Injection Spike
Diadinoxanthin
19-Hexanoyloxyfucoxanthin
Monovinyl Chl b
Peridinin
Chl c2
Prasinoxanthin
b.b carotene
Zeaxanthin
Monovinyl Chls a, a
Alloxanthin
Diatoxanthin
b.e carotene
13
SeaHARRE-3 Analysis
14
AGU/ASLO/TOS Ocean Sciences Meeting (Honolulu,
20-24 Feb 06)
To Separate or Not to Separate? That is the
HPLC Question A comparison between C18 and C8
methods J. Perl, Q. Williams, and C.
Trees Particulate absorption a pigment
reconstruction analysis Q. Williams, J. Perl,
and C. Trees