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目的建立LC-MS/MS法测定nodosin在小鼠体内的血药浓度,研究nodosin在小鼠体内的药物动力学。方法将KM小鼠单次腹腔注射nodosin生理盐水注射液,断头取血,采用液液萃取法处理血浆样品,以齐墩果酸为内标,采用ESI离子源,多重反应监测模式进行负离子检测,检测离子质荷比(m/z)为nodosin 361.0/256.9;齐墩果酸(内标)以m/z 455.2/407.2和m/z 455.2/455.2分别进行定性和定量。使用Diamonsil C18色谱柱(4.6 mm×250 mm,5μm);以甲醇-5 mmol·L-1乙酸铵水溶液为流动相,梯度洗脱;流速为1.0 m L·min-1;柱温25℃。结果小鼠腹腔注射nodosin后,nodosin血药浓度在3.9~1000 ng·m L-1范围内线性关系良好(r2>0.99),最低检测限为1.0 ng·m L-1。日内、日间精密度(RSD)均小于9.8%,方法学考察均符合生物样品分析的要求。小鼠血浆中nodosin及内标物均无内源性基质的干扰。结论本研究建立的LC-MS/MS分析方法灵敏可靠,特异性高,适用于nodosin的药动学研究。
Objective To establish a method for the determination of nodosin in mice by LC-MS / MS and to study the pharmacokinetics of nodosin in mice. Methods KM mice were injected intraperitoneally with nodosin saline for decapitation, and the plasma samples were processed by liquid-liquid extraction. Oleanolic acid was used as the internal standard, and negative ions were detected by ESI ion source and multiplex reaction monitoring (M / z) was nodosin 361.0 / 256.9. Oleanolic acid (internal standard) was qualitatively and quantitatively determined at m / z 455.2 / 407.2 and m / z 455.2 / 455.2, respectively. A Diamonsil C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase was methanol-5 mmol·L -1 ammonium acetate aqueous solution with a gradient of 1.0 mL · min-1. The column temperature was 25 ℃. Results After nodosin was intraperitoneally injected into mice, nodosin showed a good linearity (r2> 0.99) in the range of 3.9-1000 ng · m L-1 with a minimum detectable limit of 1.0 ng · m L-1. The intra-day and inter-day precision (RSD) were less than 9.8%, and the methodological studies all met the requirements of biological sample analysis. Mouse plasma nodosin and internal standard no endogenous matrix interference. Conclusion The LC-MS / MS method established in this study is sensitive, specific and suitable for pharmacokinetic studies of nodosin.