Analysis of phytochelatin (g-Glu-Cys)2-Gly by a multi-channel electrochemical detector in online connection with reversed-phase liquid chromatography
| Authors | |
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| Year of publication | 2004 |
| Type | Article in Proceedings |
| Conference | Book of Abstracts I - 55th Annual Meeting of the ISE |
| MU Faculty or unit | |
| Citation | |
| Field | Analytic chemistry |
| Keywords | Reversed-phase liquid chromatography; electrochemical detector; phytochelatin; limit detection |
| Description | Phytochelatins (PCs) are Cys-rich peptides produced by plant or some animal cells as protective molecules against heavy-metal ions in the cytoplasm. PCs have the basic formula (g-Glu-Cys)n-X (n: 2 to 11, X: terminal amino acid as Gly). PCs contain cysteine SH-groups and can form metal-PC complexes in cell environment. For the analysis of PCs different separation and detection methods were used, including electrochemical methods. In this work we present determination of PC2 using HPLC-ED. For this purpose hydrodynamic voltammetric measurements were used at a potential of 0.7 V giving the best current response. We tested the effect of temperature of the chromatographic column and the electrochemical detector on chromatographic separation. The best results were observed at 25 C. An HPLC-ED system consisted of two solvent delivery pumps and an eight-channel CoulArray electrochemical detector. This detector was located after the HPLC system with an XDB C18 reverse-phase column (mobile phase: trifluoroacetic acid/acetonitrile; flow-rate 0.4 mL/min). It was found that the PC2 limit detection was improved by the addition of ascorbic acid (in millimolar concentration) to the sample. This combination of HPLC with electrochemical detection is a suitable tool for determination of the picomolar content of PC2 on the column. |
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