UHPLC-MS/MS测定人血浆中依匹哌唑浓度及其生物等效性研究

Determination of brexpiprazole Concentration in human Plasma by UHPLC-MS/MS and Its bioequivalence study

关键词:  依匹哌唑  抗精神病药  血浆浓度  生物等效性  超高效液相色谱-串联质谱法

Key words:  brexpiprazole  antipsychotics  plasma concentration  bioequivalence  UHPLC-MS/MS

摘要:

目的 建立简便灵敏的超高效液相色谱-串联质谱法(ultra performance liquid chromatography tandem mass spectrometry，UHPLC-MS/MS)测定人血浆中依匹哌唑浓度，并应用于2种片剂的生物等效性研究。方法 采用Waters Acquity UPLC BEH C₁₈色谱柱(2.1 mm×50 mm，1.7 μm)，以0.1%甲酸水溶液(A)-乙腈-甲醇(50∶50，含0.1%甲酸) (B)梯度洗脱，流速0.4 mL·min^-1，进样量为2 μL，柱温40 ℃，以正离子MRM模式测定依匹哌唑(m/z 434.2→273.2)的浓度，依匹哌唑-d₈(m/z 442.4→281.3)作为内标，离子源为ESI源。血浆样本加入内标，加入甲醇后进行蛋白沉淀，取上清液稀释后进样检测。结果 依匹哌唑在0.2~50 ng·mL^-1呈线性关系，定量下限为0.2 ng·mL^-1，质控样品批内、批间精密度CV≤5.0%，准确度相对偏差在标示值-1.7%~5.5%内，提取回收率、专属性、基质效应、稳定性等各项指标均符合国家药品监督管理局的技术指导原则。本法被成功应用于健康受试者单剂口服2 mg依匹哌唑口崩片的生物等效性研究，参比制剂空腹及餐后给药状态下平均C_max分别为19.62和20.83 ng·mL^-1，受试制剂空腹及餐后给药状态下平均C_max分别为21.68和19.74 ng·mL^-1。结论 该方法具有前处理过程简单,灵敏度高,色谱峰形好的优点。受试制剂依匹哌唑口崩片与其参比制剂相比，具有生物等效性。

Abstract:

OBJECTIVE To establish a fast and sensitive UHPLC-MS/MS method for determination of brexpiprazole in human plasma and to investigate the bioequivalence between 2 formulations of tablets. METHODS Waters Acquity UPLC BEH C₁₈ (2.1 mm×50 mm, 1.7 μm) column was used with the mobile phase consisting of 0.1% formic acid water(A) and acetonitrile-methanol(50∶50, containing 0.1% formic acid)(B) in gradient elution. The flow rate was controlled at 0.4 mL·min^-1 with the injection volume of 2 μL and the column temperature was set at 40 ℃. A mass spectrometer equipped with electrospray ionization source was used and brexpiprazole(m/z 434.2→273.2) was monitored in positive ion MRM mode, with brexpiprazole-d₈ (m/z 442.4→281.3) as internal standard, ion source was ESI source. After addition of internal standard, plasma protein was precipitated by methanol, and supernatants were detected after dilution. RESULTS Brexpiprazole was linear in the range of 0.2-50 ng·mL^-1 and the lower limit of quantification was 0.2 ng·mL^-1. The intra-day and inter-day precision CV of quality-control samples was ≤5.0%, and the accuracy was in the range of -1.7%-5.5% in terms of relative error. Recovery rate, specificity, matrix effect and stability met the guiding principles and criteria of NMPA. The method was successfully applied to a bioequivalence study of brexpiprazole orally disintegrating tablets containing 2 mg in healthy volunteers. The average C_max under fasting and fed condition of the reference tablet were 19.62 and 20.83 ng·mL^-1. The average C_maxunder fasting and fed condition of the test tablet were 21.68 and 19.74 ng·mL^-1. CONCLUSION The method is sensitive and simple in process, and can produce well chromatographic performance. The test brexpiprazole orally disintegrating tablet is bioequivalent to the reference tablets.