Analysis of phytochelatins and other thiol-containing compounds by RP-HPLC with monobromobimane precolumn derivatization

In this article, a method for quantitative determination of phytochelatins (PC _n being the classic example) and other thiol-containing compounds in mixed standard solution and plant tissues is presented. Thiols were converted to fluorescent derivatives by precolumn derivatization with monobromobimane. The results showed that PC _n and other thiol-containing compounds in standard mixed solutions were rapidly separated within 15 min by using a ACN 0.1% trifluoroacetic acid binary gradient elution. Glutathione was representatively selected to test the precision of this method. The calibration curve was linear in the range of 1.25–160 ng μl⁻¹ (regression coefficient r²=0.9999). It was confirmed that this method was rapid, simple, highly sensitive, stable, and had the property of simultaneous determination of PC n and other thiol-containing compounds. This method was applied to determine PC _n and other thiol-containing compounds in a Cd hyperaccumulator Sedum alfredii in response to Cd. It was found that no PC _n was detected in any tissue at any Cd treatment, suggesting that Cd hyperaccumulation and detoxification in this plant is not based on PC synthesis. Copyright © 2006 Higher Education Press and Springer-Verlag.