基于全血稳定性研究的头孢哌酮/舒巴坦治疗药物监测的临床采样流程建立

Establishment of a Clinical Sampling Procedure for Therapeutic Drug Monitoring of Cefoperazone/Sulbactam Based on Whole Blood Stability Studies

关键词:  头孢哌酮  舒巴坦  全血稳定性  治疗药物监测  临床采样流程

Key words:  cefoperazone  sulbactam  whole blood stability  therapeutic drug monitoring  clinical sampling procedure

摘要:

摘要：目的 通过LC-MS/MS法测定人血浆中头孢哌酮/舒巴坦的浓度，结合全血稳定性研究制订临床采样流程，并应用到治疗药物监测个体化给药方案设计。方法 色谱柱为Dimension C18（2.1 mm×100 mm, 3 μm）；流动相为0.1%甲酸水-乙腈；梯度洗脱；柱温40 ℃；血浆样品经乙腈沉淀蛋白，取上清液加水稀释后进样分析；采用电喷雾离子源，扫描方法为多反应监测（MRM），头孢哌酮、舒巴坦、内标头孢他啶和他唑巴坦检测离子对分别为m/z 646.00→143.15，m/z 232.00→140.20，m/z 574.00→468.05和m/z 298.90→138.15；分别考察头孢哌酮/舒巴坦全血样本在不同温度下的稳定性。结果 头孢哌酮和舒巴坦的血浆浓度线性范围分别是（0.070~32.00 μg×mL-1）、（0.15~64.00 μg×mL-1），低、中、高质控样品的日内、日间精密度均小于15%，基质效应分别为100.78%~107.73%、97.55~110.28%；提取回收率分别为89.70%~102.71%、94.26%~104.70%；头孢哌酮/舒巴坦在肝素钠采血管的全血样本于23 ℃条件下24 h、4 ℃条件48 h内稳定，优于同等放置条件下EDTA-K2采血管（4 h、12 h内稳定）。结论 建立的同时测定头孢哌酮/舒巴坦的LC-MS/MS法具有灵敏度高、准确性高、重复性好等优点，能够满足临床血浆样品分析检测的要求。

Abstract:

ABSTRACT: OBJECTIVE To establish a LC-MS/MS method for the determination of concentration of cefoperazone /sulbactam in human blood and build clinical sampling process based on its stability study, then applied to therapeutic drug monitoring,which can be applied to the design of individualized dosing regimen for therapeutic drug monitoring. METHODS The chromatographic column was Dimension C18(2.1 mm×100 mm, 3 μm).The mobile phase was 0.1% formic acid water-acetonitrile.The elution method and detection column temperature were gradient elution and 40 ℃. Analysis was set up multi-reaction monitoring mode (MRM) and belectrospray ion sourc .The MRM transitions monitored for cefoperazone, sulbactam，ceftazidime and tazobactam (Internal standard) were m/z 646.00→143.15, m/z 232.0→140.20，m/z 574.0→468.05, and m/z 298.90→138.15, respectively.Plasma samples were precipitated by acetonitrile, and the supernatant was diluted with water before injection for analysis.The stability of whole blood cefoperazone/sulbactam at different temperatures were investigated. RESULTS The linear ranges of plasma concentrations of cefoperazone/sulbactam were (0.070~32.00 μg×mL-1) and (0.15~64.00 μg×mL-1), respectively. The intra-day and inter-day precision of low, medium and high quality control samples were all less than 15%, and the matrix effects were 100.78%~107.73% and 97.55~110.28%, respectively.The extraction recoveries were 89.70%~102.71% and 94.26%~104.70%, respectively. Cefoperazone/sulbactam are stable in heparin sodium blood collection tubes at 23 °C for 24 hours and 4 °C for 48 hours, which is better than the same conditions in EDTA-K2 blood collection tubes (4 hours and 12 hours of stability). CONCLUSION The LC-MS/MS method developed for the simultaneous determination of cefoperazone/sulbactam exhibits high sensitivity, accuracy, and reproducibility, thereby meeting the requirements for clinical plasma sample testing.