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

• Published in: Journal of Chromatography A 2008-10, Vol.1207 (1), p.72-83
• Main Authors: Minocha, Rakesh, Thangavel, P., Dhankher, Om Parkash, Long, Stephanie
• Format: Article
• Language: English
• Subjects: Agronomy. Soil science and plant productions; Algae; Arabidopsis; Biological and medical sciences; Cell Culture Techniques; chemical constituents of plants; Chemical constitution; Chlorophyta - chemistry; Chromatography, High Pressure Liquid - methods; Crambe; cultured cells; cysteine; Economic plant physiology; Forest trees; Fundamental and applied biological sciences. Psychology; gamma-glutamylcysteine; glutathione; high performance liquid chromatography; HPLC; mBBr; Phytochelatins; Phytochelatins - isolation & purification; Plant physiology and development; plant tissues; plants; Plants - chemistry; polythiols; Red spruce; Reproducibility of Results; Rice; Sensitivity and Specificity; Sugar maple; Sulfhydryl Compounds - analysis; Sulfhydryl Compounds - isolation & purification; thiols; trees; Trees - chemistry
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2008.08.023

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, γ-glutamylcysteine) along with polythiols (PC 2, PC 3, PC 4 and PC 5) within 23 min from a wide variety of samples. Total run time of the method is 35 min. Detection limits for thiols is 33 fmol for 10 μL 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.