毛细管电泳法测定硫酸多黏菌素B中的有关物质
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摘要
目的建立一种用毛细管电泳测定硫酸多黏菌素B中有关物质的分析方法。方法以含30mmol/L羟丙基-β-环糊精、5%异丙醇的130mmol/L三乙醇胺溶液(用磷酸调节pH至2.5)为运行缓冲液,熔融石英毛细管总长60cm,有效长度51.5cm,内径50μm,分离电压24kV,柱温25℃,进行了线性、检测限、精密度等方法学考察试验,并将该方法应用于硫酸多黏菌素B原料药及制剂的检测中。结果实现了多黏菌素B中主要组分B_1、B_2、B_3、B_(1-I)与相邻杂质的分离,对比了不同来源硫酸多黏菌素B有关物质的差异。结论本方法灵敏度较高、重复性较好,为硫酸多黏菌素B的质量控制提供了一种可行的分析方法。
Objective A capillary electrophoresis method was established for the determination of related substances in polymyxin B sulfate. Methods To improve the separation selectivity, hydroxypropyl-β-cyclodextrin(HP-β-CD) was necessary to be used as a buffer additive. The optimal separation conditions were as follows: the total length of the capillary was 60 cm(the effective length was 51.5 cm), 130 mmol/L triethanolamine-phosphate buffer containing 30 mmol/L HP-β-CD and 5% IPA, the applied voltage was 24 kV, and the column temperature was 25℃. Further, the linearity, limits of detection quantification, and the repeatability were examined. The contents of the related substances in polymyxin B of commercial samples were analyzed quantitatively according to the area normalization method. Results The CE method successfully separated the main components(B_1, B_2, B_3, B_(1-I)) and the adjacent impurities in polymyxin B. The contents of the related substances among various sources of polymyxin B were compared. Conclusion The CE method was sensitive and reproducible to control the quality of polymyxin B sulfate.
Objective A capillary electrophoresis method was established for the determination of related substances in polymyxin B sulfate. Methods To improve the separation selectivity, hydroxypropyl-β-cyclodextrin(HP-β-CD) was necessary to be used as a buffer additive. The optimal separation conditions were as follows: the total length of the capillary was 60 cm(the effective length was 51.5 cm), 130 mmol/L triethanolamine-phosphate buffer containing 30 mmol/L HP-β-CD and 5% IPA, the applied voltage was 24 kV, and the column temperature was 25℃. Further, the linearity, limits of detection quantification, and the repeatability were examined. The contents of the related substances in polymyxin B of commercial samples were analyzed quantitatively according to the area normalization method. Results The CE method successfully separated the main components(B_1, B_2, B_3, B_(1-I)) and the adjacent impurities in polymyxin B. The contents of the related substances among various sources of polymyxin B were compared. Conclusion The CE method was sensitive and reproducible to control the quality of polymyxin B sulfate.
引文
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[13]Cui A L,Hu X X,Gao Y,et al.Synthesis and bioactivity investigation of the individual components of cyclic lipopeptide antibiotics[J].J Med Chem,2018,61(5):1845-1857.
[14]Kristensen H K,Hansen S H.Separation of polymyxins by micellar electrokinetic capillary chromatography[J].JChromatogr A,1993,628(2):309-315.
[15]Kang J W,Van S A,Orwa J A,et al.Analysis of polymyxin B sulfate by capillary zone electrophoresis with cyclodextrin as additive method development and validation[J].JChromatogr A,2000,879(2):211-218.
[2]梁浩,滕峰,吉爱国.革兰阴性多药耐药菌和多黏菌素研究介绍[J].中国药学杂志,2012,47(24):2049-2051.
[3]Velkov T,Robert K D,Nation R L,et al.Pharmacology of polymyxin:new insights into an'old'class of antibiotics[J].Future Microbiol,2013,8(6):711-724.
[4]Yuan Z,Tam V H.Polymyxin B:A new strategy for multidrug-resistance Gram-negative organisms[J].Expert Opin Invest Drugs,2008,17(5):661-668.
[5]国家药典委员会.中华人民共和国药典[S].(2015年版二部).北京:化学工业出版社,2015:1323-1324.
[6]美国药典委员会.美国药典40版[S].2017:5750-5752.
[7]欧洲药典委员会.欧洲药典9.0版[S].2017:3366-3367.
[8]Govaerts C,Orwa J A,Van S A,et al.Characterization of polypeptide antibiotics of the polymyxin series by liquid chromatography electrospray ionization ion trap tandem mass spectrometry[J].J Pep Sci,2002,8(2):45-55.
[9]Govaerts C,Adams E,Schepdael A V,et al.Hyphenation of liquid chromatography to ion trap mass spectrometry to identify minor components in polypeptide antibiotics[J].Anal Bioanal Chem,2003,377(5):909-921.
[10]Bossche L V D,Schepdael A V,Chopra S,et al.Identification of impurities in polymyxin B and colistin bulk sample using liquid chromatography coupled to mass spectrometry[J].Talanta,2011,83(5):1521-1529.
[11]张含智,秦峰,刘浩.高效液相色谱质谱联用分析硫酸多黏菌素B中的未知杂质[J].中国药学杂志,2018,53(11):918-924.
[12]崔阿龙,胡辛欣,高岩,等.化学合成多黏菌素B和E单一组分及其抗菌活性研究[J].中国抗生素杂志,2017,42(3):198-202.
[13]Cui A L,Hu X X,Gao Y,et al.Synthesis and bioactivity investigation of the individual components of cyclic lipopeptide antibiotics[J].J Med Chem,2018,61(5):1845-1857.
[14]Kristensen H K,Hansen S H.Separation of polymyxins by micellar electrokinetic capillary chromatography[J].JChromatogr A,1993,628(2):309-315.
[15]Kang J W,Van S A,Orwa J A,et al.Analysis of polymyxin B sulfate by capillary zone electrophoresis with cyclodextrin as additive method development and validation[J].JChromatogr A,2000,879(2):211-218.