目的:建立液相色谱串联质谱法(LC-MS/MS)测定人血浆中他唑巴坦钠浓度,考察健康受试者单次/重复给予他唑巴坦单方(0.50 g)和与头胞噻肟钠3种组合的复方制剂(1∶6、1∶4和1∶3)时的体内药代动力学特征,从而为其临床组方安全性及应用提供参考。方法:通过蛋白沉淀法处理血浆样本,采用Waters Symmetry C18(50 mm×2.1 mm,3.5 μm)色谱柱,以甲醇-0.15%甲酸及0.3%氨水溶液(38∶62)为流动相,流速0.3 mL·min-1,进样器温度4 ℃,柱温35 ℃;多级反应负离子(MRM)模式,他唑巴坦和头孢拉定(内标)离子为m/z 298.80 → 138.00和m/z 347.80→303.70。利用建立的分析方法,测定静脉注射他唑巴坦单方和不同配比复方制剂的24名受试者给药后各时间点血浆样品中他唑巴坦浓度,并对获得的主要药代动力学参数进行计算分析。结果:他唑巴坦血浆样本线性范围为0.200～50.0 μg·mL-1;r2=0.994 8,定量下限(LOQ)为0.200 μg·mL-1,日内、日间精密度良好(RSD<15%),提取回收率为75.4%～81.0%,基质效应为91.8%～95.6%。3个剂量组的他唑巴坦药代动力学参数Cmax和AUC0-t随给药剂量增加而增加,Cmax呈线性增加,而AUC0-t和AUC0-∞值为非线性增加。结论:本研究建立了简便﹑准确﹑灵敏的LC-MS/MS检测法,单方/复方制剂以及单次/重复给药后的药代动力学研究表明,他唑巴坦在人体内无蓄积,并可为其血浆浓度检测及临床组方应用提供技术和理论支持。
Objective: To establish an LC-MS/MS method for determining the concentration of tazobactan in human plasma, The pharmacokinetic characteristics of tazobactan single prescription (0.50 g) and the compound preparation (1∶6, 1∶4 and 1∶3) combined with cicotaxime sodiumwere investigated in healthy subjects after single/repeated administration, so as to provide reference for the safety and application of its clinical formulation. Methods: Plasma samples were processed by protein precipitation on a Waters Symmetry C18 (50 mm×2.1 mm, 3.5 μm) column at 35 ℃ with an auto sampler temperature of 4 ℃. The mobile phase was methanol-0.15% formic acid and 0.3% ammonia water (38∶62). The flow rate was 0.3 mL·min-1. In multistage reactive anion (MRM) mode, tazobactam and cefradine (internal standard) ions were m/z 298.80→138.00 and m/z 347.80→303.70.the established analytical method was used,the concentrations of tazobactam in plasma samples of 24 subjects who received intravenous tazobactam monotherapy and compound preparations of different proportions were determined at each time point after administration,and the main pharmacokinetic parameters obtained were calculated and analyzed. Results: The linear relationship of tazobactan in plasma was well within the range of 0.200-50.0 μg·mL-1 (r2=0.994 8), the lower limit of quantitation was 0.20 μg·mL-1, and the intra-day and inter-day precisions RSD was less than 15%. The recoveries rate ranged from 75.4%-81.0%, and matrix effect ranged from 91.8%-95.6%. The pharmacokinetic parameters Cmax and AUC0-t of tazobactam in the three dose groups increased with the increase of dose, and Cmax showed a linear increase, while AUC0-t and AUC0-∞shows a nonlinear increase. Conclusion: The LC-MS/MS method established is very simple, accurate and sensitive, pharmacokinetics studies of single/compound formulations and single/repeated administration indicates that tazobactan had no accumulation in human body. The study could be provided to a technical and theoretical support for concentration detection in human plasma and clinical formula application.
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