Rapid quantitative determination of a collagenase inhibitor and its major metabolite by on-line liquid chromatography with ionspray tandem mass spectrometric detection
| Autor: | M. J. Madigan, S. R. Sharp, N. G. Knebel |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Quality Control
Detection limit Analyte Chromatography Chemistry Metabolite Extraction (chemistry) Valine General Chemistry Matrix Metalloproteinase Inhibitors Hydroxamic Acids Mass spectrometry Sensitivity and Specificity Mass Spectrometry Matrix (chemical analysis) Standard curve chemistry.chemical_compound Humans Enzyme Inhibitors Quantitative analysis (chemistry) Chromatography High Pressure Liquid |
| Zdroj: | Journal of Chromatography B: Biomedical Sciences and Applications. 673:213-222 |
| ISSN: | 0378-4347 |
| DOI: | 10.1016/0378-4347(95)00270-2 |
| Popis: | A rapid, sensitive and specific analytical method has been developed and validated to quantify the collagenase inhibitor N2-(2(5)-[(hydroxycarbamoyl)methyl)-4-methylvaleryl]-N1, 3-dimethyl-L-valin-amide (I) and its major metabolite (II) from plasma and urine. This method employs an automated solid-phase extraction procedure to isolate the analytes and the internal standard from the biological matrix. Reconstituted extracts were analyzed by HPLC-ionspray MS-MS. Chromatography was performed on a 4.6 mm I.D. reversed-phase guard column. The retention times of the analytes and the internal standard were approximately 1.3 min. The assay has a limit of quantification of 5 ng/ml plasma and a limit of detection of 1 ng/ml, based on 100-micro l plasma aliquots. No sample-drying step was required. The standard curves were linear form 5 to 5000 ng/ml using weighted linear regression analysis (1/y2). The intra- and inter-assay precision were better than + or - 10% with intra- and inter-assay accuracies between 95 and 105%. this new HPLC-MS-MS assay procedure for I and II in plasma and urine has proven to be specific, sensitive, accurate and robust, and is being used routinely for the analysis of I in pre-clinical and clinical trial samples. Up to 200 unknowns may be analyzed each 24 h per analyst. |
| Databáze: | OpenAIRE |
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