# Presentation: 499867

## HPLC Phytoplankton Pigment Measurements: Continuation of Climate Quality Data Records (CQDR) in Support of NASA’s Earth Science Enterprise (ESE)

**C.C. Trees and J.L. Mueller**
**Center for Hydro-Optics and Remote Sensing (CHORS)**
**San Diego State University**
**(ctrees@chors.sdsu.edu)**

## HPLC Phytoplankton Pigment Protocols

**	****Method Comparison**
**	****	****Co-elution of mono- and divinyl Chl *****a***
**	****	****	****1.  Wright *****et al.***** (1991) C****18 ****Method (SIMBIOS)**
**	****	****	****	****Channels ratio (436 & 450nm) **
**Physical separation of mono- and divinyl chl *****a***
**	****	****1.*****  *****Goericke and Repeta (1993) C****8**** Method **
**	****	****2.  **********Van Heukelem and Thomas (2001) C****8**** with**
**	****	****	****column heater Method **

**HPLC Phytoplankton Pigment Protocols**
**(Bidigare *****et al.,***** 2003; NASA/TM-2003-211621/Rev 4 - Vol. V)**

## 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

**Internal Standard**

## Divinyl & Monovinyl Chl a (664 nm)

- Divinyl & Monovinyl Chl *a* (664 nm)

- Divinyl Chl *b*

- Monovinyl Chl *a*

- Divinyl Chl *a*

**Divinyl and Monovinyl Chl *****a***** Separation**

## 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**

## HPLC Samples Requested

**HPLC Samples Requested**

## HPLC Phytoplankton Pigment Protocols

**	****	****1.  GF/F glass fiber filters (25 mm)**
**	****	****2.  Positive pressure filtration recommended**
**	****	****3.  Filtration Volumes (green as possible)**
**	****	****	****a.  Oligotrophic waters - 3-4 liters**
**	****	****	****b.  Mesotrophic waters - 1-2 liters**
**	****	****	****c.  Eutrophic waters - 0.5-1 liter**
**	****Sample Handling, Storage and Shipping**
**	****	****1.  Fold filters in half with filtered sides facing in.**
**	****	**

**HPLC Phytoplankton Pigment Protocols**

## HPLC Phytoplankton Pigment Protocols

**	****	****2.  Place in aluminum foil envelops, cryogenic vials or **
**	****	****	****HistoPrep tissue capsules.**
**3.  Label samples with a sequential number.**
**	****	****4.  Store and ship (dry shipper) in liquid nitrogen (LN****2****).**
**	****	****5.  PI’s pay for all shipping costs (includes costs **
**	****	****	****for returning LN****2**** shipping container to PI).**
**	****Recordkeeping (**_**electronically transmitted to CHORS**_**)**
**	****	****1.  For each sample the volume filtered, station number, **
**	****	****	****date, time, depth, lat and long will be sent. **

**HPLC Phytoplankton Pigment Protocols**

## Positive Pressure Filtration System

**Positive Pressure Filtration System**

## Bottle Cap Valves/Hardware

**Bottle Cap Valves/Hardware**

## Positive Pressure Filtration System

- Nalgene Polypropylene Stopcocks with PTFE Plug, Fisher (2mm #14-630-1A)
- Tee Barbed Fitting, Cole Palmer (1/4" #U06462-20)
- Elbow Barbed Fitting, Cole Palmer (1/4" #U06461-10)
- In-Line Delrin Filter Holder, VWR (25mm, Pall Gelman #28144-255)
- PVC Labcock Valve, RyanHerco (1/4" MT X H #5309.102)
- PVC Bushing N870, RyanHerco (1/2x1/4" MTXT Spear #3839S072)
- Gast Oil less Diaphragm-Type Pressure/Vacuum Pump, Fisher (#01-092-29)

**Positive Pressure Filtration System**

## Liquid Nitrogen Refrigerator (Dry Shipper)

**Liquid Nitrogen Refrigerator (Dry Shipper)**

- Taylor-Wharton Cryogenics
- Model Number 3DS
-    LN2 Capacity - 3 liters
-    *Static Hold Time - 20 days
-    *Working Time - 14 days
- Hard-shell Shipping Case
- *Nominal values
- Midwest Scientific (_[www.midsci.com](http://www.midsci.com/)__)_
- Thermolyne Arctic Express (Fisher)
- Taylor-Wharton CX Series (Cole-
-      Parmer)
- Taylor-Wharton LD Model CP30 (VWR)