Determination of tetrahydrothiophene formation as a probe of in vitro busulfan metabolism by human glutathione S-transferase A1-1: use of a highly sensitive gas chromatographic-mass spectrometric method.
Clicks: 219
ID: 70555
1999
A method for the sensitive determination of tetrahydrothiophene (THT) in cytosolic incubation mixtures was developed. Busulfan conjugation with glutathione was predominantly catalysed by glutathione S-transferase A1-1 (GST A1-1) and THT was released from the primary metabolite by alkalization. After liquid-liquid extraction using n-pentane separation and quantification of the product was performed by gas chromatography with a mass-selective detector. The method showed good sensitivity, accuracy and reproducibility with a detection limit of 2 ng ml(-1) and a limit of quantification of 5 ng ml(-1). The suitability of the method is shown for enzyme kinetic studies in human liver cytosol as well as for determination of GST A1-1 activity.
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Authors | Ritter, C A;Bohnenstengel, F;Hofmann, U;Kroemer, H K;Sperker, B; |
Journal | journal of chromatography b, biomedical sciences and applications |
Year | 1999 |
DOI | DOI not found |
URL | URL not found |
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