HPLC-MS/MS法测定米格列奈的血药浓度
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摘要
目的建立人血浆中米格列奈浓度的HPLC-MS/MS检测方法。方法采用HPLC-MS/MS法测定米格列奈的血药浓度。以那格列奈为内标,流动相A:0.2%甲酸水溶液(含5mmol·L~(-1)乙酸铵),流动相B:甲醇-乙腈=1∶1(含0.2%甲酸,5mmol·L~(-1)乙酸铵),流速为0.3ml·min-1,进样量为10μl。结果标准曲线方程分别为:R=6.89 C-0.0153(r=0.9964,权重为1/C2)。血浆中米格列奈的浓度在0.0105~2.10μg·ml~(-1)范围内与米格列奈峰面积/内标峰面积具有良好的线性关系。血浆样品中米格列奈的平均萃取回收率为(96.47±6.18)%(n=15);内标的平均萃取回收率为(95.43±3.93)%(n=15)。血浆样品中米格列奈低、中、高3个浓度的日内RSD分别为3.14%~7.28%、3.50%~5.70%、4.41%~12.68%,日间RSD分别为5.64%、7.31%、10.37%。各项稳定性试验表明该方法符合要求。结论本试验所建立的米格列奈HPLC-MS/MS测定法能专属、灵敏、准确的测定其血药浓度,可应用于米格列奈在人体内的药动学特征研究。
Objective To establish an LC-MS/MS/MS method for determining mitiglinide in human plasma.Methods HPLC-MS/MS method was adopted to determine the concentration of mitiglinide in human plasma.Nateglinide was used as the internal standard,mobile phase A consisted of 0.2%formic acid(containing 5mmol·L~(-1) ammonium acetate),while mobile phase B consisted of methanol-acetonitrile=1∶1(containing 0.2% formic acid,5mmol·L~(-1) ammonium acetate).The flow rate was 0.3ml·min~(-1),and the injection volume was 10μl.Results The standard curve equation was R =6.89 C-0.0153(r=0.9964,the weight=1/C2).When the mitiglinide concentration in plasma ranged from 0.0105 to 2.10μg·ml~(-1).There was a good linear relationship with the peak area of mitiglinide internal standard peak area.The average extraction recovery of mitiglinide in plasma samples was(96.47±6.18)%(n=15).The average extraction recovery of the internal standard was(95.43±3.93)%(n=15).The intra-day RSD ranged from 3.14% to 7.28%,from 3.50% to 5.70%,and from 4.41% to 12.68%,respectively.The inter-day RSD ranged from 5.64%,7.31 % to 10.37%.The stability tests showed that this method met the requirements.Conclusion The mitiglinide HPLC-MS/MS determination method established in this experiment can be specific,sensitive and accurate for determining the plasma concentration,which can be applied to the research of pharmacokinetics in the human body.
Objective To establish an LC-MS/MS/MS method for determining mitiglinide in human plasma.Methods HPLC-MS/MS method was adopted to determine the concentration of mitiglinide in human plasma.Nateglinide was used as the internal standard,mobile phase A consisted of 0.2%formic acid(containing 5mmol·L~(-1) ammonium acetate),while mobile phase B consisted of methanol-acetonitrile=1∶1(containing 0.2% formic acid,5mmol·L~(-1) ammonium acetate).The flow rate was 0.3ml·min~(-1),and the injection volume was 10μl.Results The standard curve equation was R =6.89 C-0.0153(r=0.9964,the weight=1/C2).When the mitiglinide concentration in plasma ranged from 0.0105 to 2.10μg·ml~(-1).There was a good linear relationship with the peak area of mitiglinide internal standard peak area.The average extraction recovery of mitiglinide in plasma samples was(96.47±6.18)%(n=15).The average extraction recovery of the internal standard was(95.43±3.93)%(n=15).The intra-day RSD ranged from 3.14% to 7.28%,from 3.50% to 5.70%,and from 4.41% to 12.68%,respectively.The inter-day RSD ranged from 5.64%,7.31 % to 10.37%.The stability tests showed that this method met the requirements.Conclusion The mitiglinide HPLC-MS/MS determination method established in this experiment can be specific,sensitive and accurate for determining the plasma concentration,which can be applied to the research of pharmacokinetics in the human body.
引文
[1]刘茜,郭孟楠,赵辉,等.液质联用法测定人血浆中米格列奈的浓度[J].沈阳药科大学学报,2012,3(1):203-207
[2]Toshiyuki T,Harumi A,Ubukata K,et al.Comparison of the rapidity of onset of the therapeutic effect between nateglinide and mitiglinide by PK/PD analysis in rats[J].Eur J Drug Metab Pharmacokinet,2012,37(1):9-15
[3]Shigeto M,Katsura M,Matsuda M,et al.Nateglinide and mitiglinide,but not sulfonylureas,induce insulin secretion through a mechanism mediated by calcium release from endoplasmic reticulum[J].Pharmacol Exp Ther,2007,322(1):1-7
[4]Sunaga Y,Gonoi T,Shibasaki T,et al.The effects of mitiglinide(KAD-1229),a new anti-diabetic drug,on ATPsensitive K+channels and insulin secretion:comparison with the sulfonylureas and nateglinide[J].Eur J Pharmacol,2001,431(1):119-125
[5]Comaschi M,Coscelli C,Cucinotta D,et al.Cardiovascular risk factors and metabolic control in type 2diabetic subjects attending outpatient clinics in Italy:The SFIDA(survey of risk factors in Italian diabetic subjects by AMD)study[J].Nutr Metab Cardiovasc Dis,2005,15(3):204-211
[6]钟大放.以加权最小二乘法建立生物分析标准曲线的若干问题[J].药物分析杂志,1996,16(5):343-346
[7]王鹏,蒋学华,王凌.LC-MSn应用于生物样品检测中基质效应的评价[J].中国新药杂志,2011,20(20):1953-1956
[8]李鹏飞,刘丽宏,马萍,等.LC-MS/MS法测定人血浆中米格列奈浓度[J].质谱学报,2008,30(2):94-98
[9]过林,胡公允,余慧,等.LC-MS/MS测定人血浆米格列奈浓度及药代动力学研究[J].临床研究,2011,14(1):1308-1313
[2]Toshiyuki T,Harumi A,Ubukata K,et al.Comparison of the rapidity of onset of the therapeutic effect between nateglinide and mitiglinide by PK/PD analysis in rats[J].Eur J Drug Metab Pharmacokinet,2012,37(1):9-15
[3]Shigeto M,Katsura M,Matsuda M,et al.Nateglinide and mitiglinide,but not sulfonylureas,induce insulin secretion through a mechanism mediated by calcium release from endoplasmic reticulum[J].Pharmacol Exp Ther,2007,322(1):1-7
[4]Sunaga Y,Gonoi T,Shibasaki T,et al.The effects of mitiglinide(KAD-1229),a new anti-diabetic drug,on ATPsensitive K+channels and insulin secretion:comparison with the sulfonylureas and nateglinide[J].Eur J Pharmacol,2001,431(1):119-125
[5]Comaschi M,Coscelli C,Cucinotta D,et al.Cardiovascular risk factors and metabolic control in type 2diabetic subjects attending outpatient clinics in Italy:The SFIDA(survey of risk factors in Italian diabetic subjects by AMD)study[J].Nutr Metab Cardiovasc Dis,2005,15(3):204-211
[6]钟大放.以加权最小二乘法建立生物分析标准曲线的若干问题[J].药物分析杂志,1996,16(5):343-346
[7]王鹏,蒋学华,王凌.LC-MSn应用于生物样品检测中基质效应的评价[J].中国新药杂志,2011,20(20):1953-1956
[8]李鹏飞,刘丽宏,马萍,等.LC-MS/MS法测定人血浆中米格列奈浓度[J].质谱学报,2008,30(2):94-98
[9]过林,胡公允,余慧,等.LC-MS/MS测定人血浆米格列奈浓度及药代动力学研究[J].临床研究,2011,14(1):1308-1313