采用试验设计理念建立并优化丙酸交沙霉素有关物质分析方法
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摘要
目的建立丙酸交沙霉素HPLC有关物质分析方法。方法针对大环内酯类抗生素杂质谱复杂的特点,基于试验设计理念,采用响应曲面法建立了丙酸交沙霉素有关物质分析方法。确定流动相中的有机相比例、缓冲盐浓度和柱温、缓冲盐浓度作为作为优化中显著影响的自变量因素,主峰和主峰前色谱峰的分离度、第一区间杂质个数、第二区间杂质个数为优化中的因变量。结果确定丙酸交沙霉素最优化色谱系统的条件为:流动相为15mmol醋酸铵溶液-乙腈(34:66, V/V)、柱温40℃、流速1.0mL/min、色谱柱为Xtimate C_(18)(250mm×4.6mm, 5μm)。结论采用所建立方法测定丙酸交沙霉素有关物质,各杂质峰分离较好,优于现行药典方法和文献方法,且方法学验证指标较好。
Objective To develop an HPLC method to analyze the related substances in josamycin propionate.Methods Due to the complex constitutions of macrolides, the response surface method was used to develop the HPLC method of josamycin propionate based on the design of experiments concept. The organic phase content in mobile phase, the concentration of buffer, and column temperature were selected as significant effects in the design,and resolution of the main peak and the nearest peak in front and numbers of peaks were selected as dependent variables. Results The optimal condition of josamycin propionate HPLC system was as follows: the mobile phase was 15 mmol ammonium acetate-acetonitrile(34:66); the column temperature was 40℃; the flow rate was 1.0 mL/min and the column used was Xtimate C_(18)(250 mm×4.6 mm, 5μm). Conclusion Using the developed method, all peaks were separated well from each other, which was better than methods in pharmacopoeias and literatures. Besides, the validation parameters all met the specifications of standards.
Objective To develop an HPLC method to analyze the related substances in josamycin propionate.Methods Due to the complex constitutions of macrolides, the response surface method was used to develop the HPLC method of josamycin propionate based on the design of experiments concept. The organic phase content in mobile phase, the concentration of buffer, and column temperature were selected as significant effects in the design,and resolution of the main peak and the nearest peak in front and numbers of peaks were selected as dependent variables. Results The optimal condition of josamycin propionate HPLC system was as follows: the mobile phase was 15 mmol ammonium acetate-acetonitrile(34:66); the column temperature was 40℃; the flow rate was 1.0 mL/min and the column used was Xtimate C_(18)(250 mm×4.6 mm, 5μm). Conclusion Using the developed method, all peaks were separated well from each other, which was better than methods in pharmacopoeias and literatures. Besides, the validation parameters all met the specifications of standards.
引文
[1]Bossche L V D,Daidone F,Schepdael A V,et al.Characterization of impurities in josamycin using dual liquid chromatography combined with mass spectrometry[J].JPharm Biomed Anal,2013,73(2):66-76.
[2]姚尚辰,胡昌勤.中国药典2015年版大环内酯类抗生素的拟增修订重点[J].中国抗生素杂志,2018,43(4):373-379.
[3]Orlandini S,Pinzauti S,Furlanetto S.Application of quality by design to the development of analytical separation methods[J].Anal Bioanal Chem,2013,405(2-3):443-450.
[4]Govaerts C,Chepkwony H K,Van Schepdael A,et al.Application of liquid chromatography-ion trap mass spectrometry to the characterization of the 16-membered ring macrolide josamycin propionate[J].J Mass Spectrom,2004,39(4):437-446
[5]Chepkwony H K,Rodriguez E,Govaerts C,et al.Development of a liquid chromatography method for the assay and purity testing of josamycin propionate[J].J Sep Sci,2003,26(6-7):489-495.
[6]Roets E,Lepoudre X,Van Rompaey V,et al.Liquid chromatography of josamycin propionate on poly(styrenedivinylbenzene)[J].J Chromatogr A,1998,812(1-2):303-308.
[2]姚尚辰,胡昌勤.中国药典2015年版大环内酯类抗生素的拟增修订重点[J].中国抗生素杂志,2018,43(4):373-379.
[3]Orlandini S,Pinzauti S,Furlanetto S.Application of quality by design to the development of analytical separation methods[J].Anal Bioanal Chem,2013,405(2-3):443-450.
[4]Govaerts C,Chepkwony H K,Van Schepdael A,et al.Application of liquid chromatography-ion trap mass spectrometry to the characterization of the 16-membered ring macrolide josamycin propionate[J].J Mass Spectrom,2004,39(4):437-446
[5]Chepkwony H K,Rodriguez E,Govaerts C,et al.Development of a liquid chromatography method for the assay and purity testing of josamycin propionate[J].J Sep Sci,2003,26(6-7):489-495.
[6]Roets E,Lepoudre X,Van Rompaey V,et al.Liquid chromatography of josamycin propionate on poly(styrenedivinylbenzene)[J].J Chromatogr A,1998,812(1-2):303-308.