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To establish a high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method for analyzing the concentrations of methotrexate (MTX) and its metabolite 7-hydroxymethotrexate (7-OH-MTX) in human plasma, and to use it for clinical drug monitoring, in order to explore the relationship between their concentrations and liver and kidney toxicity.
Acetonitrile precipitation protein method was used to preprocess the samples, combined with HPLC-MS/MS detection. 369 blood samples from pediatric and adult patients who received high-dose methotrexate (HDMTX) chemotherapy were collected for methodological validation, and Spearman correlation analysis between concentration and hepatotoxicity and nephrotoxicity was performed using statistical software SPSS 27.
The linear relationship of MTX in human plasma was good within the concentration range of 0.005-2 μmol·L-1 (r=0.998 8), and the linear relationship of 7-OH-MTX was good within the concentration range of 0.05-20 μmol·L-1 (r=0.997 4). The intra batch and inter batch precision were all less than 10%, the extraction recovery rate reached 87.0-106.1%, and the coefficient of variation of matrix effect was less than 7%. The analysis of blood concentration measurements at 42 hours, 72 hours, and after HDMTX infusion showed that the concentration ratio of 7-OH-MTX/MTX was significantly negatively correlated with liver function indicators alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (P<0.01), while there was no significant correlation with renal function indicators creatinine (CR) and creatinine clearance rate (CCR) (P>0.05).
The established method is simple, accurate, and sensitive, and is suitable for clinical drug therapy monitoring. By monitoring 7-OH-MTX/MTX, the occurrence of liver toxicity can be effectively predicted for timely clinical drug rescue.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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