伏立康唑对映体HPLC手性拆分及分析
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摘要
目的:建立伏立康唑对映体HPLC手性分离及分析方法。方法:以β-CD及其衍生物为流动相手性添加剂,探讨β-CD衍生物类型及浓度、流动相中有机相体积分数、色谱柱固定相类型等因素对伏立康唑对映体HPLC手性拆分效果的影响,在此基础上,建立伏立康唑对映体的HPLC分析方法,并进行方法学考察。结果:β-CD衍生物中,SBE-β-CD适宜作为伏立康唑对映体HPLC手性拆分添加剂,以甲醇-水(45∶55)为流动相,磷酸二氢钾浓度为15 mmol.L-1,SBE-β-CD浓度为10.0 mmol.L-1时,伏立康唑对映体在C8及C18固定相上均能得到有效拆分。在上述条件下,以Hypersil C18色谱柱(4.6 mm×150 mm,5μm)分析伏立康唑对映体,流动相流速为1.0 mL.min-1,伏立康唑对映体能实现基线分离,伏立康唑浓度在31.2～156μg.mL-1范围内,浓度与峰面积呈良好线性关系(R2=0.9998),伏立康唑加样回收率大于98%,稳定性及重现性RSD分别为0.77%,0.67%。结论:建立了一种伏立康唑对映体的手性拆分和分析方法,可为伏立康唑的分离与分析提供借鉴。
Objective: To establish an HPLC chiral separation and analysis method of voriconazole enantiomers.Methods: Using β-CD and its derivates as chiral additives of mobile phase,the influences of the type and concentration of cyclodextrins,mobile phase ratio,and type of column stationary phase on the chiral separation were investigated.Then an HPLC analysis method of voriconazole enantiomers was established and methodologically discussed.Results: Sulfobutylether-β-cyclodextrins(SBE-β-CD),as the chiral additive,is more suitable for HPLC separation of voriconazole enantiomers than other β-CD derivates,and voriconazole enantiomers could be effectively separated on C8 and C18 stationary phase with methanol-water(45:55)as mobile phase at the KH2PO4,SBE-β-CD concentrations of 15.0 mmol·L-1 and 10.0 mmol·L-1,respectively.Under the conditions mentioned above,the voriconazole enantiomers was base-line separated on the Hypersil ODS C18 column(4.6 mm×150 mm,5 μm)at a mobile phase flow rate of 1.0 mL·min-1.By methodological analysis,the average recovery of voriconazole was more than 98%,and the RSD values of stability and reproducibility were 0.77% and 0.67%,respectively.Conclusion: The proposed method is suitable for the chiral separation and analysis of voriconazole enantiomers by HPLC.
Objective: To establish an HPLC chiral separation and analysis method of voriconazole enantiomers.Methods: Using β-CD and its derivates as chiral additives of mobile phase,the influences of the type and concentration of cyclodextrins,mobile phase ratio,and type of column stationary phase on the chiral separation were investigated.Then an HPLC analysis method of voriconazole enantiomers was established and methodologically discussed.Results: Sulfobutylether-β-cyclodextrins(SBE-β-CD),as the chiral additive,is more suitable for HPLC separation of voriconazole enantiomers than other β-CD derivates,and voriconazole enantiomers could be effectively separated on C8 and C18 stationary phase with methanol-water(45:55)as mobile phase at the KH2PO4,SBE-β-CD concentrations of 15.0 mmol·L-1 and 10.0 mmol·L-1,respectively.Under the conditions mentioned above,the voriconazole enantiomers was base-line separated on the Hypersil ODS C18 column(4.6 mm×150 mm,5 μm)at a mobile phase flow rate of 1.0 mL·min-1.By methodological analysis,the average recovery of voriconazole was more than 98%,and the RSD values of stability and reproducibility were 0.77% and 0.67%,respectively.Conclusion: The proposed method is suitable for the chiral separation and analysis of voriconazole enantiomers by HPLC.
引文
1 Butters M,Ebbs J,Green SP,et al.Process development of voricon-azole:a novel broad-spectnm trizole antifungal agent.Org ProcessRes Dev,2001,5(1):28
2CHANG Sheng(昌盛),DONG Jin-hua(董金华).Voriconazole(伏立康唑).Chin J Med Chem(中国药物化学杂志),2007,17(3):197
3 Owens PK,Fell AF,Coleman MW,et al.Separation of the voricon-azole stereoisomers by capillary electrophoresis and liquid chromatog-raphy.Enantiomer,1999,4(2):79
4 Owens PK,Fell AF,Coleman MW,et al.Complexation of vori-conazole stereoisomers with neutral and anionic derivatised cy-clodextrins.J Incl Phenom Macrocycl Chem,2000,38(1-4):133
5 Ferretti R,Gallinella B,La Torre F,et al.Direct resolution of a newantifungal agent,voriconazole(UK-109,496)and its potential im-purities,by use of coupled achiral-chiral high-performance liquidchromatography.Chromatographia,1998,47(11/12):649
6WANG Yong-zhong(汪永忠).Separation for chiralcel enantiomerof voriconazole by HPLC(伏立康唑对映体的手性高效液相色谱分离).China Pharm(中国药师),2006,9(3):231
7FENG Xiao-ming(冯小明),ZENG De-chao(曾德超),JIANGYao-zhong(蒋耀忠).A novel method of resolution-chiral inclu-sion complexation(一种新的手性拆分方法-包结拆分).Chin JOrg Chem(有机化学),2000,20(2):131
8XIE Jian-wei(谢剑炜),YANG Zao-ping(杨造萍),RUAN Jin-xiu(阮金秀).Chiral recognition of cyclodextrin supramolecular sys-tem chiral separation of enantiomer drugs by reversed phase HPLCwith cyclodextrin as a mobile phase and stationary phase(手性药物对映体的环糊精手性流动相、手性固定相HPLC法拆分).ActaPharm Sin(药学学报),1998,33(2):143
9NING Feng-rong(宁凤容),HUANG Ke-long(黄可龙),JIAO Fei-peng(焦飞鹏),et al.Thermodynamic study on en-antioresolution of chlorthalidone by high performance liquid chro-matography usingβ-cyclodextrin and its derivatives as chiral ad-ditives(β-环糊精及其衍生物手性添加剂HPLC法拆分氯噻酮对映体过程热力学研究).Chin J Anal Lab(分析试验室),2006,25(8):1
10 Armstrong DW,Nome F,Spino LA,et al.Efficient detection and e-valuation of cyclodextrin multiple complex formation.J Am ChemSoc,1986,108(7):1418
11WANG Zhong-hui(王钟辉),MAN Rui-lin(满瑞林),TANGKe-wen(唐课文).Studies on the retention and separation mecha-nisms of phenylsuccinic acid enantiomers with HP-β-CD as chrialmobile phase additive in HPLC(HP-β-CD对苯基琥珀酸对映体色谱保留机制及拆分机理的研究).J Instrum Anal(分析测试学报),2009,28(2):162
12CHEN De-ying(陈德英),CHEN Yu-ying(陈玉英),HU Yu-zhu(胡玉筑).Study on chiral selective inclusion and retention char-acteristics of cis-trans isomers and enantiomers of sertraline withHP-β-CD as a mobile phase modifier(HP-β-环糊精对盐酸舍曲林异构体手性选择包合和色谱保留行为影响的研究).ChinJ Chromatogr(色谱),2004,22(6):595
2CHANG Sheng(昌盛),DONG Jin-hua(董金华).Voriconazole(伏立康唑).Chin J Med Chem(中国药物化学杂志),2007,17(3):197
3 Owens PK,Fell AF,Coleman MW,et al.Separation of the voricon-azole stereoisomers by capillary electrophoresis and liquid chromatog-raphy.Enantiomer,1999,4(2):79
4 Owens PK,Fell AF,Coleman MW,et al.Complexation of vori-conazole stereoisomers with neutral and anionic derivatised cy-clodextrins.J Incl Phenom Macrocycl Chem,2000,38(1-4):133
5 Ferretti R,Gallinella B,La Torre F,et al.Direct resolution of a newantifungal agent,voriconazole(UK-109,496)and its potential im-purities,by use of coupled achiral-chiral high-performance liquidchromatography.Chromatographia,1998,47(11/12):649
6WANG Yong-zhong(汪永忠).Separation for chiralcel enantiomerof voriconazole by HPLC(伏立康唑对映体的手性高效液相色谱分离).China Pharm(中国药师),2006,9(3):231
7FENG Xiao-ming(冯小明),ZENG De-chao(曾德超),JIANGYao-zhong(蒋耀忠).A novel method of resolution-chiral inclu-sion complexation(一种新的手性拆分方法-包结拆分).Chin JOrg Chem(有机化学),2000,20(2):131
8XIE Jian-wei(谢剑炜),YANG Zao-ping(杨造萍),RUAN Jin-xiu(阮金秀).Chiral recognition of cyclodextrin supramolecular sys-tem chiral separation of enantiomer drugs by reversed phase HPLCwith cyclodextrin as a mobile phase and stationary phase(手性药物对映体的环糊精手性流动相、手性固定相HPLC法拆分).ActaPharm Sin(药学学报),1998,33(2):143
9NING Feng-rong(宁凤容),HUANG Ke-long(黄可龙),JIAO Fei-peng(焦飞鹏),et al.Thermodynamic study on en-antioresolution of chlorthalidone by high performance liquid chro-matography usingβ-cyclodextrin and its derivatives as chiral ad-ditives(β-环糊精及其衍生物手性添加剂HPLC法拆分氯噻酮对映体过程热力学研究).Chin J Anal Lab(分析试验室),2006,25(8):1
10 Armstrong DW,Nome F,Spino LA,et al.Efficient detection and e-valuation of cyclodextrin multiple complex formation.J Am ChemSoc,1986,108(7):1418
11WANG Zhong-hui(王钟辉),MAN Rui-lin(满瑞林),TANGKe-wen(唐课文).Studies on the retention and separation mecha-nisms of phenylsuccinic acid enantiomers with HP-β-CD as chrialmobile phase additive in HPLC(HP-β-CD对苯基琥珀酸对映体色谱保留机制及拆分机理的研究).J Instrum Anal(分析测试学报),2009,28(2):162
12CHEN De-ying(陈德英),CHEN Yu-ying(陈玉英),HU Yu-zhu(胡玉筑).Study on chiral selective inclusion and retention char-acteristics of cis-trans isomers and enantiomers of sertraline withHP-β-CD as a mobile phase modifier(HP-β-环糊精对盐酸舍曲林异构体手性选择包合和色谱保留行为影响的研究).ChinJ Chromatogr(色谱),2004,22(6):595