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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Vande Casteele, Sofie R. F. Mulliez, Sylvie M. N. Boussery, Koen De Bock, Lies Van Bocxlaer, Jan F. P. |
| Description | Country affiliation: Belgium Author Affiliation: De Bock L ( Laboratory of Medical Biochemistry and Clinical Analysis, Ghent University, Harelbekestraat 72, 9000 Gent, Belgium. Lies.DeBock@UGent.be) |
| Abstract | For the determination of the in vitro cytochrome P450 activity in microsomes, a quantification method for the probe metabolites, formed during incubation, is required. Due to insufficient sensitivity of a previously developed high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for some of the metabolites, a fast and easy derivatization method with pyridine-3-sulfonyl chloride (PS) is described. Acetaminophen (CYP1A2), dextrorphan (CYP2D6), hydroxy-chlorzoxazone (CYP2E1) and hydroxy-mephenytoin (CYP2C19) can be derivatized because of the presence of a phenolic OH, whereas hydroxy-midazolam (CYP3A4) and hydroxy-tolbutamide (CYP2C9) remain unchanged. As PS improves the ionization efficiency in the positive electrospray ionization (ESI) mode, the sensitivity of the detection is improved significantly and meets requirements for the activity determination. Native negative electrospray type molecules, moreover, become positive ESI candidates. The direct derivatization in the aqueous incubation medium, without any other sample pre-treatment steps, such as evaporation or extraction, makes this procedure easy to perform. The method using 20s microwave irradiation was shown to equal a 10min reaction in a 60°C heating block, consequently simplifying and shortening the process. Collision induced fragmentation of the derivatives resulted in at least one native compound, rather than derivative, specific product ion, thereby improving the selectivity of the method in the multiple reaction monitoring mode. The HPLC-MS/MS method was validated, and was demonstrated to be sensitive, selective, precise and accurate. The absence of a relative matrix effect was established, notwithstanding that an absolute matrix effect was observed. The analysis of a sample after microsomal incubation, from which some of the metabolites could not be quantified using the method without derivatization, proved the usefulness of the method. |
| ISSN | 00219673 |
| Issue Number | 6 |
| Volume Number | 1218 |
| e-ISSN | 18733778 |
| Journal | Journal of Chromatography A |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2011-02-11 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Chromatography, High Pressure Liquid Methods Cytochrome P-450 Enzyme System Metabolism Pharmaceutical Preparations Analysis Pyridines Chemistry Sulfinic Acids Tandem Mass Spectrometry Animals Drug Stability Humans Least-squares Analysis Male Rats Rats, Wistar Reproducibility Of Results Sensitivity And Specificity Journal Article Research Support, Non-u.s. Gov't Discipline Analytical Chemistry |
| Content Type | Text |
| Resource Type | Article |
| Subject | Organic Chemistry Medicine Analytical Chemistry Biochemistry |
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