# Presentation: 678109

## 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**
**(ctrees@chors.sdsu.edu)**
**(jperl@chors.sdsu.edu)**

## HPLC Phytoplankton Pigment Protocols

**	****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)**

## HPLC Phytoplankton Pigment Protocols

**	****	****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 (LN****2****).**
**	****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

**Template for Ancillary Data**

## HPLC System Calibrations 

**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).

## Fluorometer 

**Fluorometer **
**Calibration**

## Number of Samples 

**by Date and PI.**

## Number of Samples Analyzed by PI

**Number of Samples Analyzed by PI**

## Number of Samples Pending

**Number of Samples Pending**

## Vitamin E Internal Standard (284 nm)

- Vitamin E Internal Standard (284 nm)

- Zeaxanthin

- Divinyl Chl *b*

- Divinyl Chl *a*

- Monovinyl Chl *a*

- Vitamin E

-  Carotene

**Problems - Vitamin E Internal Standard**

-  trans*-****-apo-8'-carotenal *(Fluka Chemical Company)

##   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**

## Culture Mixture (450 nm)

- Culture Mixture (450 nm)

- Chl *c**3*

- Injection Spike

- Peridinin

- Alloxanthin

- Fucoxanthin

- . carotene

- Monovinyl Chl *a*

- Diatoxanthin

- 19’-Hexanoyloxyfucoxanthin

- Zeaxanthin

- Monovinyl Chl *b*

- Monovinyl Chls *a’, a”*

- Chl *c**2*

- Diadinoxanthin

- Prasinoxanthin

- . carotene

**Mixture of Pigments from Cultures**

## SeaHARRE-3 Analysis

## AGU/ASLO/TOS Ocean Sciences Meeting

***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)**