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| Content Provider | Springer Nature Link |
|---|---|
| Author | Spadaro, A. Bousquet, E. Santagati, N.A. Vittorio, F. Ronsisvalle, G. |
| Copyright Year | 2005 |
| Abstract | Here, we provide an extremely simple, rapid, direct, sensitive and specific method to quantify, for the first time in human colon carcinoma cells (HT29) and epidermoid human larynx carcinoma cells (HEp-2), the intracellular glutathione (GSH) using an electrochemical detector coupled to a high performance liquid chromatograph (HPLC-ECD). GSH determination in cancer cells are of valuable interest because GSH has been implicated in the development of resistance to several anticancer drugs. Separation of GSH was performed on a Alltima C18 column using a binary pre-mixed mobile phase of methanol-NaH2PO4 (10 mM, pH 3.0 with phosphoric acid) (2:98, v/v). GSH was detected with an oxidation potential of +500 mV. The limit of detections was 0.30 pMol and good linearity (r = 0.9988) was found. The intra-day and the inter-day recoveries were 97.1–101.3% and 98.2–101.8% respectively, whereas the accuracies were less than 2.56% and 3.02% respectively. The usefulness of the method was established by subjecting HT29 and HEp-2 cells to an oxidant stress generated by menadione, a substance able to decrease GSH level in cells. The experiments show that the HPLC-ECD method developed could represent an useful alternative to the existing procedures since its simplicity and rapidity. |
| Starting Page | 11 |
| Ending Page | 16 |
| Page Count | 6 |
| File Format | |
| ISSN | 00095893 |
| Journal | Chromatographia |
| Volume Number | 62 |
| Issue Number | 1-2 |
| e-ISSN | 16121112 |
| Language | English |
| Publisher | Vieweg Verlag |
| Publisher Date | 2005-06-27 |
| Publisher Place | Wiesbaden |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Column liquid chromatography Electrochemical detection Cell cultures (HT29 and HEp-2) Glutathione Analytical Chemistry Organic Chemistry Pharmacy Biochemistry Plant Sciences Measurement Science, Instrumentation |
| Content Type | Text |
| Resource Type | Article |
| Subject | Organic Chemistry Analytical Chemistry Biochemistry Clinical Biochemistry |
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