基于万古霉素手性柱的马来酸扑尔敏对映体分离及其识别机理
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摘要
药物是环境介质中一类新型污染物,多数药物具有手性对映体特性,药物对映体的分离分析是研究对映体水平药物环境行为和效应的前提.采用商品手性柱Chirobiotic®V,在反相条件下,对马来酸扑尔敏进行了手性分离,考察了不同流速、柱温、pH值和流动相组成对分离的影响,并优化了色谱条件.结果表明:以含0.1%(体积分数)冰醋酸和0.1%(体积分数)三乙胺的水溶液(TEAA)与四氢呋喃(THF)混合为流动相,在所研究范围内,随着流动相中THF含量的增加、pH值增大和流速增大,马来酸扑尔敏对映体的分离效果降低;柱温影响存在峰值现象,即分离效果随温度的升高先增大后减小.优化得到的拆分条件为:流动相TEAA∶THF为95∶5(V/V),pH值为3,流速为0.3 mL·min~(-1),柱温为15℃.利用分子对接技术模拟万古霉素不同区域与扑尔敏对映体间的相互作用,计算得到了最稳定结合能,据此预测对映体的出峰顺序与实验结果完全一致.初步手性识别机理研究结果表明,引起手性识别的主要作用力为氢键作用.
Pharmaceuticals are a large class of emerging pollutants in environmental matrix. It has been reported that more than 50% of these pharmaceuticals are chiral compounds. Thus, the chiral separation and analysis of pharmaceuticals are prerequisites for investigating environmental behavior and impact of drug enantiomers. The commercial vancomycin-bonded chiral stationary phase(CSP) combined with high performance liquid chromatography(HPLC) in reverse mode was used for the enantioseparation of chlorphenamine maleate enantiomers. The flow rate, column temperature, pH value and composition of mobile phase were optimized for the effective separation. The results showed that when TEAA containing 0.1% of glacial acetic acid and 0.1% of triethylamine(in volume ratio) was mixed with tetrahydrofuran(THF) as mobile phase, the reduced enantioresolution and selectivity were observed with an increased THF percentage, pH value and flow rate. The effect of column temperature on separation indicated that maximum values in enantioresolution and selectivity occurred at 15 ℃. As a result, the optimal chromatographic condition was: TEAA∶THF=95∶5(V/V), pH value of 3, flow rate of 0.3 mL·min~(-1), temperature of 15 ℃. Using molecular docking technique and binding energy calculations to simulate the interaction processes between chlorphenamine maleate and vancomycin, the magnitudes of steady combination energy(ΔG) was obtained, which coincided with experimental elution order of the enantiomers. The molecular docking results showed that the main retention mechanisms of chiral chlorphenamine maleate onto vancomycin were hydrogen bonding.
Pharmaceuticals are a large class of emerging pollutants in environmental matrix. It has been reported that more than 50% of these pharmaceuticals are chiral compounds. Thus, the chiral separation and analysis of pharmaceuticals are prerequisites for investigating environmental behavior and impact of drug enantiomers. The commercial vancomycin-bonded chiral stationary phase(CSP) combined with high performance liquid chromatography(HPLC) in reverse mode was used for the enantioseparation of chlorphenamine maleate enantiomers. The flow rate, column temperature, pH value and composition of mobile phase were optimized for the effective separation. The results showed that when TEAA containing 0.1% of glacial acetic acid and 0.1% of triethylamine(in volume ratio) was mixed with tetrahydrofuran(THF) as mobile phase, the reduced enantioresolution and selectivity were observed with an increased THF percentage, pH value and flow rate. The effect of column temperature on separation indicated that maximum values in enantioresolution and selectivity occurred at 15 ℃. As a result, the optimal chromatographic condition was: TEAA∶THF=95∶5(V/V), pH value of 3, flow rate of 0.3 mL·min~(-1), temperature of 15 ℃. Using molecular docking technique and binding energy calculations to simulate the interaction processes between chlorphenamine maleate and vancomycin, the magnitudes of steady combination energy(ΔG) was obtained, which coincided with experimental elution order of the enantiomers. The molecular docking results showed that the main retention mechanisms of chiral chlorphenamine maleate onto vancomycin were hydrogen bonding.
引文
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Mojtahedi M M,Chalavi S,Ghassempour A,et al.2007.Chiral separation of three agrochemical toxins enantiomers by high-performance liquid chromatography on a vancomycin crystalline degradation products-chiral stationary phase[J].Biomedical Chromatography,21(3):234-240
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Raverdy V,Ampe E,Hecq J D,et al.2013.Stability and compatibility of vancomycin for administration by continuous infusion[J].Journal of Antimicrobial Chemotherapy,68(5):1179-1182
Ribeiro A R,Maia A S,Cass Q B,et al.2014.Enantioseparation of chiral pharmaceuticals in biomedical and environmental analyses by liquid chromatography:an overview[J].Journal of Chromatography B,968:8-21
Shen J,Wei H,Yu X,et al.2011.Chiral separation of chlorpheniramine by HPLC with β-Cyclodextrin nitro group derivative as stationary phase[J].Journal of South-Central University for Nationalities (Natural Science Edition),30(4):32-35
Sun J Y,Xing Y P,Song J X,et al.2015.Chiral separation of 4 drugs by capillary electrophoresis[J].Journal of Shenyang Pharmaceutical University,32:199-203
Svensson L A,D?nnecke J,Karlsson K E,et al.2000.Polar organic phase liquid chromatography with packed capillary columns using a vancomycin chiral stationary phase[J].Chirality,12(8):606-613
王世玉.1998.世界手性药物选集[M].北京:国家医药管理局信息中心.234-246
Zhang D,Xu X,Ma S.2015.The study of enantioseparation of zolmitriptan on vancomycin-bonded chiral stationary phase[J].Journal of Separation Science,28(18):2501-2504
AlHarthy F.2009.Separation of aryl nitro-compounds by HPLC on monolithic columns[D].Loughborough: Loughborough University.32-34
Armstrong D W,Tang Y,Chen S,et al.1994.Macrocyclic antibiotics as a new class of chiral selectors for liquid chromatography[J].Analytical Chemistry,66(9):1473-1484
Bagnall J P,Evans S E,Wort M T,et al.2012.Using chiral liquid chromatography quadrupole time-of-flight mass spectrometry for the analysis of pharmaceuticals and illicit drugs in surface and wastewater at the enantiomeric level[J].Journal of Chromatography A,1249(15):115-129
Bosáková Z,Curínová E,Tesarová E.2005.Comparison of vancomycin-based stationary phases with different chiral selector coverage for enantioselective separation of selected drugs in high-performance liquid chromatography[J].Journal of Chromatography A,1088(1):94-103
程燕,贾欣欣,刘健艺,等.2012.以CM-β-CD为HPLC手性流动相添加剂分离扑尔敏等6种手性药物对映体[J].沈阳药科大学学报,29(2):121-125
丁国生,刘莺,丛润滋,等.2004.去甲万古霉素键合手性固定相拆分β-受体阻滞剂类药物及其结构类似物的研究[J].色谱,22(4):386-389
Evans S E,Davies P,Lubben A,et al.2015.Determination of chiral pharmaceuticals and illicit drugs in wastewater and sludge using microwave assisted extraction,solid-phase extraction and chiral liquid chromatography coupled with tandem mass spectrometry[J].Analytica Chimica Acta,882(30):112-126
Gasper M P,Berthod A,Nair U B,et al.1996.Comparison and modeling study of vancomycin,ristocetin A,and teicoplanin for CE enantioseparations[J].Analytical Chemistry,68(15):2501-2514
胡昌勤,洪建文.2009.毛细管电泳中万古霉素拆分氧氟沙星对映体手性识别位点的探讨[J].药学学报,44(8):905-910
胡曼玲.2003.卫生化学(第5版)[M].北京:人民卫生出版社.56-59
Kasprzyk-Hordern B.2011.ChemInform abstract:Pharmacologically active compounds in the environment and their chirality[J].Cheminform,42(8):4466-4503
Kasprzykhordern B,Kondakal V V,Baker D R.2010.Enantiomeric analysis of drugs of abuse in wastewater by chiral liquid chromatography coupled with tandem mass spectrometry[J].Journal of Chromatography A,1217(27):4575-4586
Knox J H,Scott H P.1983.B and C terms in the Van Deemter equation for liquid chromatography[J].Journal of Chromatography A,282(DEC):297-313
Liu Y,Gu X H.2011.Pharmacology of Chiral Drugs// Lin G Q,You Q D,Cheng J F.Chiral Drugs:Chemistry and Biological Action[M].Hoboken:John Wiley & Sons,Inc.323-345
Ma R,Wang B,Lu S,et al.2016a.Characterization of pharmaceutically active compounds in Dongting Lake,China:Occurrence,chiral profiling and environmental risk[J].Science of the Total Environment,557-558:268-275
Ma R,Wang B,Yin L,et al.2016b.Characterization of pharmaceutically active compounds in Beijing,China:Occurrence pattern,spatiotemporal distribution and its environmental implication[J].Journal of Hazardous Materials,323(Pt A):147-155
Mojtahedi M M,Chalavi S,Ghassempour A,et al.2007.Chiral separation of three agrochemical toxins enantiomers by high-performance liquid chromatography on a vancomycin crystalline degradation products-chiral stationary phase[J].Biomedical Chromatography,21(3):234-240
Morris G M,Goodsell D S,Halliday R S,et al.2015.Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function[J].Journal of Computational Chemistry,19(14):1639-1662
Osterberg F,Morris G M,Sanner M F,et al.2002.Automated docking to multiple target structures:incorporation of protein mobility and structural water heterogeneity in AutoDock[J].Proteins Structure Function & Bioinformatics,46(1):34-40
Raverdy V,Ampe E,Hecq J D,et al.2013.Stability and compatibility of vancomycin for administration by continuous infusion[J].Journal of Antimicrobial Chemotherapy,68(5):1179-1182
Ribeiro A R,Maia A S,Cass Q B,et al.2014.Enantioseparation of chiral pharmaceuticals in biomedical and environmental analyses by liquid chromatography:an overview[J].Journal of Chromatography B,968:8-21
Shen J,Wei H,Yu X,et al.2011.Chiral separation of chlorpheniramine by HPLC with β-Cyclodextrin nitro group derivative as stationary phase[J].Journal of South-Central University for Nationalities (Natural Science Edition),30(4):32-35
Sun J Y,Xing Y P,Song J X,et al.2015.Chiral separation of 4 drugs by capillary electrophoresis[J].Journal of Shenyang Pharmaceutical University,32:199-203
Svensson L A,D?nnecke J,Karlsson K E,et al.2000.Polar organic phase liquid chromatography with packed capillary columns using a vancomycin chiral stationary phase[J].Chirality,12(8):606-613
王世玉.1998.世界手性药物选集[M].北京:国家医药管理局信息中心.234-246
Zhang D,Xu X,Ma S.2015.The study of enantioseparation of zolmitriptan on vancomycin-bonded chiral stationary phase[J].Journal of Separation Science,28(18):2501-2504