modis science team meeting baltimore, md (4-6 jan 06) hplc phytoplankton pigment measurements:...

MODIS Science Team MeetingBaltimore, MD (4-6 Jan 06)

HPLC Phytoplankton Pigment Measurements: Continuation of Climate Quality Data Records (CQDR) in Support

of NASA’s Earth Science Enterprise (ESE)

Chuck Trees and Jason Perl

Center for Hydro-Optics and Remote Sensing (CHORS)

San Diego State University([email protected])([email protected])


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

HPLC Phytoplankton Pigment Protocols

(Bidigare et al., 2003; NASA/TM-2003-211621/Rev 4 - Vol. V)


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.

HPLC Phytoplankton Pigment Protocols


Template for Ancillary Data

PI Ancillary Data Format12/21/05 jrp

general general general mm/dd/yy hh:mm number number general number number numberPI PI's ID# Cruise Date GMT Station# Depth (m) Geographic Location LAT (dec) LONG (dec) VolFilt (mL)

Chuck Trees Haw1 MOBY 05/01/04 14:45 6 0 Hawaii 21.333 -158.000 1250Chuck Trees Haw2 MOBY 05/01/04 14:45 6 5 Hawaii 21.333 -158.000 1250Chuck Trees Haw3 MOBY 05/01/04 14:45 6 10 Hawaii 21.333 -158.000 1250Chuck Trees Haw4 MOBY 05/01/04 16:05 7 0 Hawaii 21.456 -158.200 1000Chuck Trees Haw5 MOBY 05/01/04 16:05 7 20 Hawaii 21.456 -158.200 1000Chuck Trees Haw6 MOBY 05/01/04 16:05 7 40 Hawaii 21.456 -158.200 1000

OR

Chuck Trees 1 MOBY 05/01/04 14:45 6 0 Hawaii 21.333 -158.000 1250Chuck Trees 2 MOBY 05/01/04 14:45 6 5 Hawaii 21.333 -158.000 1250Chuck Trees 3 MOBY 05/01/04 14:45 6 10 Hawaii 21.333 -158.000 1250Chuck Trees 4 MOBY 05/01/04 16:05 7 0 Hawaii 21.456 -158.200 1000Chuck Trees 5 MOBY 05/01/04 16:05 7 20 Hawaii 21.456 -158.200 1000Chuck Trees 6 MOBY 05/01/04 16:05 7 40 Hawaii 21.456 -158.200 1000


HPLC System Calibrations

Date ChlorophyllsPhaeopigmentsCarotenoids1-Nov-04 3 -- - 51-Dec-04 2 -- - -- -

15-Dec-04 5 3 171-J an-05 2 -- - -- -

15-J an-05 5 3 1731-J an-05 2 -- - -- -1-Mar-05 5 3 171-Apr- 05 5 3 141-May-05 2 -- - 21-J ul-05 5 3 14

15-Jul -05 2 -- - -- -1-Aug-05 2 -- - -- -1-Sep-05 2 -- - -- -

15-Sep-05 2 -- - -- -

145 pigment compound calibration curves with 6-10 injections per curve (870-1,450 std injections or

24-40 days of continuous running standards).


Digital Turner Calibration

0.0

0.5

1.0

1.5

2.0

2.5

Sep-02 Mar-03 Oct-03 Apr-04 Nov-04 May-05 Dec-05

Date

Calibration Factor (mg m**-3/ Fluorescence)

0.0

0.5

1.0

1.5

2.0

2.5

Tau Factor

Calibration Factor

Tau Factor

Linear (Tau Factor)

Linear (Calibration Factor)

Fluorometer Calibration


Number of Samples by Date and PI.

PI Analysis Date Number of Samples G. Mitchell Dec-04 173R. Letelier Dec-04 210

D. McGillicuddy Dec-04 290

D. McGillicuddy Jan-05 170M. Moline Jan-05 359M. Moline Jan-05 184Glenn Cota Jan-05 285A. Mannino Mar-05 99D. Siegel Mar-05 253

D. McGillicuddy Apr-05 230G. Mitchell Apr-05 232H. Dierssen Apr-05 111

A. Subramaniam Apr-05 49

D. Stramski Apr-05 48A. Subramaniam May-05 12

D. Stramski May-05 11

M. Moline Jun-05 198H. Dierssen Jun-05 58N. Nelson Jun-05 27

A. Subramaniam Jun-05 19A. Subramaniam Jul-05 54

G. Mitchell Jul-05 274A. Mannino Jul-05 114M. Moline Jul-05 322

SeaHARRE-3 Jul-05 164M. Moline Aug-05 462G. Mitchell Aug-05 138

D. McGillicuddy Aug-05 262A. Subramaniam Sep-05 9

F. Chavez Sep-05 250D. McGillicuddy Sep-05 353MODIS Total 5,390


Number of Samples Analyzed by PI

PI # of Samples Moline 1,525

McGillicuddy/Siegel 1,275Mitchell 817

Cota 285Siegel 253

Chavez 250Mannino 213Letelier 210Dierssen 169

SeaHARRE-3 164Subramaniam 143

Stramski 59Nelson 27

MODIS Total 5,390McGillicuddy 600

Clark 423

TOTAL 6,413


Number of Samples Pending

PI # of SamplesMannino 200

Siegel 215Clark 27

Stramski 48Stramski 180

TOTAL 670


Minutes

2 4 6 8 10 12 14 16 18 20 22 24 26

mAU

0

25

50

75

100

125

150

mAU

0

25

50

75

100

125

150

PDA-284nm20040702007 Vitamin E Internal Standard

(284 nm)Vitamin E Internal Standard

(284 nm)

Zeaxan

thin

Div

inyl C

hl b

Div

inyl C

hl a

Mon

ovin

yl C

hl a

Vit

am

in E

Caro

ten

e

Problems - Vitamin E Internal Standard trans--apo-8'-carotenal (Fluka Chemical Company)


1. 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.

1. 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


Minutes

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28

mAU

0

10

20

30

40

50

60

mAU

0

10

20

30

40

50

60PDA-450nm20040701002 Culture Mixture

(450 nm)Culture Mixture

(450 nm)

Ch

l c 3

Inje

ctio

n S

pik

e

Peri

din

in

Allo

xan

thin

Fuco

xan

thin

.

caro

ten

e

Mon

ovin

yl C

hl a

Dia

toxan

thin

19

’-H

exan

oylo

xyfu

coxan

thin

Zeaxan

thin

Mon

ovin

yl C

hl b

Mon

ovin

yl C

hls

a’,

a”

Ch

l c 2

Dia

din

oxan

thin

Pra

sin

oxan

thin

.

caro

ten

e

Mixture of Pigments from Cultures


SeaHARRE-3 AnalysisSH3 HPLC Calculations12/20/05 jrp Total [Tot Access Pigs]

[Chl a] w/o chlaSH_CENT# DUP SH3-ID# % DV a (mg m-3) CV (mg m-3) CV

SH3_Cent013 a 01a 10.75% 0.3931 0.4212SH3_Cent028 a 01b 11.55% 0.4177 6.35 0.4086 4.21SH3_Cent066 a 01c 11.00% 0.4462 0.4438SH3_Dup_Cent013 b 01a 10.43% 0.3402 0.3795SH3_Dup_Cent028 b 01b 11.56% 0.3840 8.47 0.3821 4.49SH3_Dup_Cent066 b 01c 11.11% 0.4020 0.4111





“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

AGU/ASLO/TOS Ocean Sciences Meeting(Honolulu, 20-24 Feb 06)

modis science team meeting baltimore, md (4-6 jan 06) hplc phytoplankton pigment measurements:...

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