Here we provide an extremely simple, rapid, direct, sensitive and specific method to quantify for the first time intracellular GSH in human prostate cancer cells (DU145) and human lung adenocarcinoma cells (A549) using an electrochemical detector coupled to a high performance liquid chromatograph (HPLC-ECD). GSH determination in cancer cells are importance because GSH has been involved 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 methanolNaH2PO4 (10 mM, pH 3.0 with phosphoric acid) (2:98 v/v). GSH was detected with an oxidation potential of + 500 mVolt. The limit of detections was 0.33 pMol. Linearity, accuracy and precision were found to be acceptable. The experiments show that the HPLC-ECD method developed in this study could represent an useful alternative to the existing procedures since its simplicity and rapidity.

Rapid Analysis of Glutathione in Human Prostate Cancer Cells (DU145) and Human Lung Adenocarcinoma Cells (A549) by HPLC with Electrochemical Detection

SPADARO, Angelo;
2011

Abstract

Here we provide an extremely simple, rapid, direct, sensitive and specific method to quantify for the first time intracellular GSH in human prostate cancer cells (DU145) and human lung adenocarcinoma cells (A549) using an electrochemical detector coupled to a high performance liquid chromatograph (HPLC-ECD). GSH determination in cancer cells are importance because GSH has been involved 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 methanolNaH2PO4 (10 mM, pH 3.0 with phosphoric acid) (2:98 v/v). GSH was detected with an oxidation potential of + 500 mVolt. The limit of detections was 0.33 pMol. Linearity, accuracy and precision were found to be acceptable. The experiments show that the HPLC-ECD method developed in this study could represent an useful alternative to the existing procedures since its simplicity and rapidity.
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/20.500.11769/69017
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 4
  • ???jsp.display-item.citation.isi??? ND
social impact