Pronounced and differential effects of ionic strength and pH on testosterone oxidation by membrane-bound and purified forms of rat liver microsomal cytochrome P-450
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Cited by (37)
Direct coupling of electromembrane extraction to mass spectrometry - Advancing the probe functionality toward measurements of zwitterionic drug metabolites
2017, Analytica Chimica ActaCitation Excerpt :The mentioned dynamic EME setups directly coupled to MS [18–20] make use of EME to monitor dynamic compositional changes of analytes in an enzymatic reaction chamber. Such enzymatic reactions like the conversion of parent drugs into drug metabolites by cytochrome P450 enzymes however have to be performed at physiological pH values making the extraction process for certain reaction products inefficient [23]. Compounds that are zwitterionic at physiological pH are such a class of compounds that cannot be extracted efficiently as they carry both positive and negative charges.
Simultaneous measurement of CYP1A2 activity, regioselectivity, and coupling: Implications for environmental sensitivity of enzyme-substrate binding
2011, Archives of Biochemistry and BiophysicsCitation Excerpt :Buffer concentration did not alter the ratio of several testosterone metabolites from purified rat CYP2A1, CYP2B1, and CYP2C11 [14] or of midazolam metabolites from human liver microsomes [16], consistent with our findings that regioselectivity was insensitive to buffer concentration. However, a change in regioselectivity with pH was reported for the oxidation of testosterone by rat CYP2A1, CYP2B1, and CYP2C11 [14] and for cyclosporine metabolism of human CYP3A4 [15]. Likewise, our results revealed a change in regioselectivity with pH and additionally with buffer type.
Hypotheses on how selection for some traits in rodents led to correlated responses in offspring sex ratios
2004, Journal of Theoretical BiologyDetermination of testosterone and its metabolites using liquid chromatography with elevated column temperature and flow-rate gradient
2002, Analytica Chimica ActaCitation Excerpt :The regio- and stereospecific hydroxylation of the steroid nucleus provides a sensitive indicator for the identification of a specific CYP-450 isozyme [3–8]. Testosterone metabolism has been widely used to study distribution and enzyme activity of biological tissues and cell cultures [5–11]. The most commonly used technique for analysis of testosterone and its metabolites is high performance liquid chromatography (HPLC) with isocratic elution or mobile phase gradient [3,11–15].