Separation and quantification of monothiols and phytochelatins from a wide variety of cell cultures and tissues of trees and other plants using high performance liquid chromatography

Rakesh Minocha; P Thangavel; Om Parkash Dhankher; Stephanie Long

doi:10.1016/j.chroma.2008.08.023

Separation and quantification of monothiols and phytochelatins from a wide variety of cell cultures and tissues of trees and other plants using high performance liquid chromatography

J Chromatogr A. 2008 Oct 17;1207(1-2):72-83. doi: 10.1016/j.chroma.2008.08.023. Epub 2008 Aug 13.

Authors

Rakesh Minocha¹, P Thangavel, Om Parkash Dhankher, Stephanie Long

Affiliation

¹ USDA Forest Service, Northern Research Station, Durham, NH 03824, USA. rminocha@unh.edu

PMID: 18760414
DOI: 10.1016/j.chroma.2008.08.023

Abstract

The HPLC method presented here for the quantification of metal-binding thiols is considerably shorter than most previously published methods. It is a sensitive and highly reproducible method that separates monobromobimane tagged monothiols (cysteine, glutathione, gamma-glutamylcysteine) along with polythiols (PC(2), PC(3), PC(4) and PC(5)) within 23min from a wide variety of samples. Total run time of the method is 35min. Detection limits for thiols is 33fmol for 10microlL injection. This method will be applicable to study the metal detoxification mechanisms for a wide variety of cell cultures and tissues of plants and trees including algae, Arabidopsis, crambe, rice, and red spruce.

MeSH terms

Cell Culture Techniques
Chlorophyta / chemistry
Chromatography, High Pressure Liquid / methods*
Phytochelatins / isolation & purification*
Plants / chemistry*
Reproducibility of Results
Sensitivity and Specificity
Sulfhydryl Compounds / analysis*
Sulfhydryl Compounds / isolation & purification*
Trees / chemistry

Substances

Sulfhydryl Compounds
Phytochelatins