Mistri Hiren N, Jangid Arvind G, Shrivastav Pranav S
Department of Chemistry, School of Sciences, Gujarat University, Navrangpura, Ahmedabad 380009, India; Accutest Research Lab, Satellite, Ahmedabad 380015, India.
Accutest Research Lab, Satellite, Ahmedabad 380015, India.
J Pharm Biomed Anal. 2007 Sep 21;45(1):97-106. doi: 10.1016/j.jpba.2007.06.003. Epub 2007 Jun 9.
A simple and rapid liquid chromatography/tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the simultaneous quantitation of antidiabetic drugs metformin and glyburide in human plasma using glimepiride as internal standard (IS). After acidic acetonitrile-induced protein precipitation of the plasma samples, metformin, glyburide and IS were chromatographed on reverse phase C18 (50 mm x 4.6 mm i.d., 5 microm) analytical column. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique and operating in multiple reaction monitoring (MRM) and positive ion mode. The total chromatographic run time was 3.5 min and calibration curves were linear over the concentration range of 20-2500 ng/ml for metformin and 5-500 ng/ml for glyburide. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy and precision, dilution integrity and stability studies. The recoveries obtained for the analytes and IS (>or=69%) were consistent and reproducible. Inter-batch and intra-batch coefficient of variation across four validation runs (LLOQ, LQC, MQC and HQC) was less than 8%. The accuracy determined at these levels was within +/-8% in terms of relative error (RE). The method was applied to a bioequivalence study of 500 mg metformin and 5mg of glyburide tablet after oral administration to 28 healthy human subjects under condition of fasting.
已开发并验证了一种简单快速的液相色谱/串联质谱（LC-MS/MS）方法，以格列美脲作为内标（IS），同时定量测定人血浆中的抗糖尿病药物二甲双胍和格列本脲。血浆样品经酸性乙腈诱导蛋白沉淀后，二甲双胍、格列本脲和内标在反相C18（50 mm×4.6 mm内径，5微米）分析柱上进行色谱分离。在采用电喷雾电离技术并以多反应监测（MRM）和正离子模式运行的三重四极杆质谱仪上进行定量分析。总色谱运行时间为3.5分钟，二甲双胍的校准曲线在20 - 2500 ng/ml浓度范围内呈线性，格列本脲的校准曲线在5 - 500 ng/ml浓度范围内呈线性。该方法在选择性、灵敏度、回收率、线性、准确度和精密度、稀释完整性及稳定性研究方面均得到验证。分析物和内标的回收率（≥69%）一致且可重现。在四次验证运行（LLOQ、LQC、MQC和HQC）中，批间和批内变异系数均小于8%。在这些水平下测定的准确度，相对误差（RE）在±8%以内。该方法应用于28名健康人类受试者在空腹条件下口服500 mg二甲双胍和5 mg格列本脲片剂的生物等效性研究。
