UPLC-MS/MS测定人血浆中吗啡、羟考酮、哌替啶和芬太尼的浓度研究

Simultaneous Determination of Plasma Concentrations of Morphine, Oxycodone, Pethidine and Fentanyl by UPLC-MS/MS

目的 建立同时测定人血浆中吗啡、羟考酮、哌替啶及芬太尼4种治疗药物浓度的方法,并用于治疗药物的监测。方法 血浆样本经乙腈沉淀蛋白后,以氘代吗啡作为内标,应用超高效液相色谱-串联质谱法(UPLC-MS/MS)测定药物浓度。以Agilent Eclipse XDB-C18柱(2.1 mm×50.0 mm, 1.7μm)为色谱柱,甲醇-水(2.0 mmol·L醋酸铵)为流动相等度洗脱,流速为0.3 mL·min^(-1),电喷雾离子源,应用多重反应监测模式进行正离子扫描分析。结果 吗啡、羟考酮、哌替啶及芬太尼的血药浓度在50.0~10 000.0 ng·mL^(-1)内线性关系良好,日内及日间精密度均小于15.0%,提取回收率及基质效应均在85.0%~115.0%内,稳定性等符合要求。结论 该方法灵敏、快速、专属性强,可以用于吗啡、羟考酮、哌替啶及芬太尼治疗药物浓度监测及药动学等方面的研究。

OBJECTIVE To establish a UPLC-MS/MS method to simultaneously determine the concentrations of morphine, oxycodone, pethidine and fentanyl in human plasma for therapeutic drug monitoring(TDM). METHODS Morphine-D3 was used as the isotope internal standard. The analytes were extracted from plasma by using acetonitrile and separated using an Agilent Eclipse XDB-C18 column(2.1×50 mm, 1.7 μm) with water containing 2 mmol·Lammonium acetate and methanol as the mobile phase. The flow rate was 0.3 mL·min^(-1). With positive ion electrospray ionization, the analytes were monitored on a triple quadrupole mass spectrometer in MRM mode. RESULTS Calibration curves were obtained over the concentration ranges of 50.0-10 000.0 ng·mL^(-1)for all the analytes, and the intra-and inter-day precision RSDs were <15.0%. The recovery and matrix effect were within the range of 85.0% to 115.0%. Stability was in line with relevant requirements.CONCLUSION The method is proved to be sensitive, rapid, and can be used for TDM and pharmacokinetic studies of morphine, oxycodone, pethidine, and fentanyl.