HPLC-MS/MS法测定依卡倍特钠原料药中潜在遗传毒性杂质
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摘要
为了定量测定依卡倍特钠中潜在遗传毒性杂质依卡倍特磺酸乙酯(杂质Ⅰ),参考文献方法合成依卡倍特磺酸乙酯。采用高分辨质谱、结合二级质谱与核磁共振确定其相对分子质量及化学结构。采用Thermo C18色谱柱,以5 mmol/L乙酸铵溶液(甲酸调p H至3.0)为流动相A;乙腈为流动相B,按照梯度:0 min 50%B,4 min 50%B,12 min 80%B,16 min 80%B,16.1 min 50%B,20 min 50%B进行洗脱,柱温40℃;采用电喷雾负离子化-MS/MS选择反应监测。杂质Ⅰ的线性质量浓度范围为4~150 ng/m L,且线性关系良好(r=0.999);最低定量限为4 ng/m L;杂质Ⅰ的进样精密度、重复性和加标回收率良好,耐用性良好。方法操作简便,灵敏度高,可用于依卡倍特钠原料药中潜在遗传毒性杂质依卡倍特磺酸乙酯的含量测定。
To develop a sensitive analytical method for the determination of the genotoxic impurity mono ethyl ester of ecabet( Imp-I),an HPLC-MS/MS technique was employed. Imp-I was synthesized according to the previous literatures. MS/MS and NMR were used to confirm the structure of Imp-I. A Thermo C18 column was used for chromatographic separations. The mobile phase consisting of A: 5 mmol/L ammonium acetate( p H adjusted to 3. 0 with formic acid) and B: acetonitrile,with a gradient program: 0 min 50% B,4 min 50% B,12 min 80% B,16 min80% B,16. 1 min 50% B and 20 min 50% B. The column was maintained at 40 ° C throughout the analysis. All measurements were carried out with the mass spectrometer operated under the negative ESI mode. The selective reaction monitor( SRM) transition was used. Good linearity was obtained for Imp-I over the concentration range of4-150 ng/m L with the coefficient of determination( r) of 0. 999. And the LOQ was 4 ng/m L. A rapid and sensitive HPLC-ESI-MS/MS method was developed for quantitative analysis of Imp-I in ecabet sodium APIs. This method can be of used for quality assurance of ecabet sodium in bulk commercial drugs.
To develop a sensitive analytical method for the determination of the genotoxic impurity mono ethyl ester of ecabet( Imp-I),an HPLC-MS/MS technique was employed. Imp-I was synthesized according to the previous literatures. MS/MS and NMR were used to confirm the structure of Imp-I. A Thermo C18 column was used for chromatographic separations. The mobile phase consisting of A: 5 mmol/L ammonium acetate( p H adjusted to 3. 0 with formic acid) and B: acetonitrile,with a gradient program: 0 min 50% B,4 min 50% B,12 min 80% B,16 min80% B,16. 1 min 50% B and 20 min 50% B. The column was maintained at 40 ° C throughout the analysis. All measurements were carried out with the mass spectrometer operated under the negative ESI mode. The selective reaction monitor( SRM) transition was used. Good linearity was obtained for Imp-I over the concentration range of4-150 ng/m L with the coefficient of determination( r) of 0. 999. And the LOQ was 4 ng/m L. A rapid and sensitive HPLC-ESI-MS/MS method was developed for quantitative analysis of Imp-I in ecabet sodium APIs. This method can be of used for quality assurance of ecabet sodium in bulk commercial drugs.
引文
[1]Looker AR,Ryan MP,Neubertlangille BJ,et al.Risk assessment of potentially genotoxic impurities within the framework of quality by design[J].Org Process Res Dev,2010,14(4):1032-1036.
[2]Ma L,Ma YN,Chen Z,et al.Structural alerts of genotoxic impurities[J].Chin New Drugs J(中国新药杂志),2014,23(18):2106-2111.
[3]Wu YF,Luan Y,Qi XM,et al.Advance in study on early and rapid assays of genotoxicity[J].Drug Eval Res(药物评价研究),2011,34(4):244-250.
[4]Wang XJ,Ma J.Research advances of new techniques and methods for drug safety evaluation[J].Chin J Pharm(中国医药工业杂志),2017,48(3):341-350.
[5]Wichard JD.In silico prediction of genotoxicity[J].Food Chem Toxicol,2017,106:595-599.
[6]Bhavani KG,Krishna KBM,Srinivasu N,et al.Determination of genotoxic impurity in atazanavir sulphate drug substance by LC-MS[J].J Pharm Biomed Anal,2017,132:156-158.
[7]Wollein U,Schramek N.Simultaneous determination of alkyl mesilates and alkyl besilates in finished drug products by direct injection GC/MS[J].Eur J Pharm Sci,2012,45(1/2):201-204.
[8]Grigori K,Loukas YL,Malenovi'c A,et al.Chemometrically assisted development and validation of LC-MS/MS method for the analysis of potential genotoxic impurities in meropenem active pharmaceutical ingredient[J].J Pharm Biomed Anal,2017,145:307-314.
[9]Zhou J,Xu J,Zheng XY,et al.Determination of methyl methanesulfonate and ethyl methanesulfonate in methanesulfonic acid by derivatization followed by high-performance liquid chromatography with ultraviolet detection[J].J Sep Sci,2017,40(17):3414-3421.
[10]Ito Y,Onoda Y,Nakamura S,et al.Effects of the new anti-ulcer drug ecabet sodium(TA-2711)on pepsin activity.II.Interaction with substrate protein[J].Jpn J Pharmacol,1993,62(2):175-181.
[11]Ito Y,Shibata K,Hongo A,et al.Ecabet sodium,a locally acting antiulcer drug,inhibits urease activity of Helicobacter pylori[J].Eur J Pharmacol,1998,345(2):193-198.
[12]Murata H,Kawano S,Tsuji S,et al.Combination therapy of ecabet sodium and cimetidine compared with cimetidine alone for gastric ulcer:prospective randomized multicenter study[J].J Gastroenterol Hepatol,2003,18(9):1029-1033.
[13]Liu GJ,Pei ZD,Zhang JX,et al.Determination of contents and related substances of ecabet sodium granules by HPLC[J].Chin J Mod Appl Pharm(中国现代应用药学),2012,29(1):70-72.
[14]Zhang D,Du X,Liu M,et al.Determination of ecabet in human plasma by high-performance liquid chromatography-tandem mass spectrometry[J].J Chromatogr B,2008,863(2):223-228.
[15]Ruan XL,Zheng XY,Xu J,et al.Advances in analytical techniques for the determination of genotoxic impurities in pharmaceuticals[J].J China Pharm Univ(中国药科大学学报),2016,47(3):267-274.
[2]Ma L,Ma YN,Chen Z,et al.Structural alerts of genotoxic impurities[J].Chin New Drugs J(中国新药杂志),2014,23(18):2106-2111.
[3]Wu YF,Luan Y,Qi XM,et al.Advance in study on early and rapid assays of genotoxicity[J].Drug Eval Res(药物评价研究),2011,34(4):244-250.
[4]Wang XJ,Ma J.Research advances of new techniques and methods for drug safety evaluation[J].Chin J Pharm(中国医药工业杂志),2017,48(3):341-350.
[5]Wichard JD.In silico prediction of genotoxicity[J].Food Chem Toxicol,2017,106:595-599.
[6]Bhavani KG,Krishna KBM,Srinivasu N,et al.Determination of genotoxic impurity in atazanavir sulphate drug substance by LC-MS[J].J Pharm Biomed Anal,2017,132:156-158.
[7]Wollein U,Schramek N.Simultaneous determination of alkyl mesilates and alkyl besilates in finished drug products by direct injection GC/MS[J].Eur J Pharm Sci,2012,45(1/2):201-204.
[8]Grigori K,Loukas YL,Malenovi'c A,et al.Chemometrically assisted development and validation of LC-MS/MS method for the analysis of potential genotoxic impurities in meropenem active pharmaceutical ingredient[J].J Pharm Biomed Anal,2017,145:307-314.
[9]Zhou J,Xu J,Zheng XY,et al.Determination of methyl methanesulfonate and ethyl methanesulfonate in methanesulfonic acid by derivatization followed by high-performance liquid chromatography with ultraviolet detection[J].J Sep Sci,2017,40(17):3414-3421.
[10]Ito Y,Onoda Y,Nakamura S,et al.Effects of the new anti-ulcer drug ecabet sodium(TA-2711)on pepsin activity.II.Interaction with substrate protein[J].Jpn J Pharmacol,1993,62(2):175-181.
[11]Ito Y,Shibata K,Hongo A,et al.Ecabet sodium,a locally acting antiulcer drug,inhibits urease activity of Helicobacter pylori[J].Eur J Pharmacol,1998,345(2):193-198.
[12]Murata H,Kawano S,Tsuji S,et al.Combination therapy of ecabet sodium and cimetidine compared with cimetidine alone for gastric ulcer:prospective randomized multicenter study[J].J Gastroenterol Hepatol,2003,18(9):1029-1033.
[13]Liu GJ,Pei ZD,Zhang JX,et al.Determination of contents and related substances of ecabet sodium granules by HPLC[J].Chin J Mod Appl Pharm(中国现代应用药学),2012,29(1):70-72.
[14]Zhang D,Du X,Liu M,et al.Determination of ecabet in human plasma by high-performance liquid chromatography-tandem mass spectrometry[J].J Chromatogr B,2008,863(2):223-228.
[15]Ruan XL,Zheng XY,Xu J,et al.Advances in analytical techniques for the determination of genotoxic impurities in pharmaceuticals[J].J China Pharm Univ(中国药科大学学报),2016,47(3):267-274.