LC-MS/MS法测定盐酸噻吩诺啡鼻腔喷雾剂的生物利用度

Study on bioavailability of thenorphine hydrochloride nasal spray in rat using liquid chromatography-mass/mass spectrometry

目的:建立大鼠血浆中盐酸噻吩诺啡的液质联用测定法(LC-MS/MS法),比较盐酸噻吩诺啡片剂和鼻腔喷雾剂的生物利用度。方法:血浆中加入内标丁丙诺啡后甲醇沉淀蛋白。流动相:乙腈-0.01%甲酸,梯度洗脱,流速为0.5 mL.min-1,柱后分流。色谱柱:Agilent-C18柱(150 mm×2.1 mm,5 mm)。离子化方式:电喷雾离子化。多反应监测(multiple reaction monitoring,MRM):定量离子对,噻吩诺啡m/z522.5→m/z97.1,内标丁丙诺啡为m/z468.5→m/z396.2。结果:盐酸噻吩诺啡鼻腔喷雾剂、片剂的绝对生物利用度分别(70.84±51.16)%和(1.22±0.16)%,鼻喷剂的生物利用度极明显高于片剂(>58倍),其血药峰浓度明显大于口服给药。结论:建立了简单快速、特异性强且灵敏度高的LC-MS/MS法,用于盐酸噻吩诺啡的体内定量分析。盐酸噻吩诺啡鼻喷剂显著提高了其在大鼠体内的生物利用度,为临床应用提供一种新的剂型选择。

Objective： To establish a liquid chromatography-mass/mass spectrometry（LC-MS/MS） method for the determination of thenorphine hydrochloride in rat plasma and to compare the bioavailability of thenorphine hydrochloride tablet（suspension） and thenorphine hydrochloride nasal spray.Methods： Buprenorphine was the internal standard.The blood samples were extracted by methanol.Separation was performed on an ODS-C18 column（150 mm×2.1 mm,5 μm,Agilent,USA） with a mobile phase consisting of acetonitrile-0.01% formic acid at flow rate of 0.5 mL·min^-1.Detection was performed by a mass/mass spectrometer in Multiple Reaction Monitoring mode via ESI interface.Sample was monitored with the protonated ion currents at m/z=522.5m/z=97.1 for thenorphine hydrochloride and m/z=468.5m/z=396.2 for buprenorphine.Results： The Absolute bioavailability of nasal spray was（70.84±51.16）%,while conventional tablet was（1.22±0.16）%.The relative bioavailability of nasal spray was as 58 times as that of conventional tablet.Conclusion： A simple,sensitive and selective method was developed to determine simultaneously the concentrations of thenorphine hydrochloride with different formulations in rat plasma by LC-MS/MS method.The thenorphine hydrochloride nasal spray was confirmed with a higher relative bioavailability than conventional tablet,which would be provided as a new choice for the clinical application.