色谱技术在中药制剂质量控制中的应用研究
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摘要
中药制剂的质量优劣是使其迈向国际市场的关键因素之一,而中药制剂的质量判定也是使国外中药制剂准入国内市场的重要环节,这些都对中药制剂的质量控制提出了很高要求。采用色谱技术对中药制剂的有效成分和有害成分进行分析与测试是进行质量控制的可行方法。
本论文在查阅大量文献的基础上,对复方丹参滴丸和固经丸的质量控制方法进行改进。主要内容包括三个部分。首先采用毛细管气相色谱法及反相高效液相色谱法分别测定了复方丹参滴丸中冰片的含量。冰片具有挥发性,因此气相色谱法测冰片含量的方法较为成熟。但是由于冰片无特征紫外吸收,所以直接采用高效液相色谱检测比较困难。根据冰片结构中含羟基的特点,可采用高效液相色谱衍生法检测。3,5-二硝基苯甲酰氯作为柱前衍生化试剂,在叔胺或吡啶类化合物的催化下可反应生成具强紫外吸收的3,5-二硝基苯甲酸冰片酯。故将两种方法分别用于测定复方丹参滴丸中冰片含量,以验证后者的准确性。冰片的平均回收率为101.2%,RSD为3.1%。供试品溶液在8h内稳定。高效液相色谱法测定的实验结果表明,在1～10nmol范围内,色谱峰面积与进样量呈线性关系。复方丹参滴丸中冰片的回收率为99.8%,RSD为1.6%(n=4)。对比发现,两种方法所测含量相当,均可用于测定冰片含量。
其次,采用反相高效液相色谱法测定了固经丸中有效成分芍药苷的含量,改进了该中药制剂的质量控制方法。芍药苷在0.44～3.96μg范围内的线性关系良好(r=0.9999),平均加样回收率为99.2%(n=5),重现性试验RSD为0.82%(n=5)。供试品溶液在24h内稳定性良好。结果表明高效液相色谱法可用于含有效成分芍药苷的中药制剂的质量控制。
最后以毛细管气相色谱法测定了复方丹参滴丸中有机氯农药的残留量。实验结果表明,11种农药在1.0×10~(-4)～5.0×10~(-1)μg/ml范围内的线性关系良好,相关系数在0.9988～0.9999范围之间。平均回收率在74.2～101.5%(n=6)范围内,RSD在2.4～8.7%之间,该方法可用于中药制剂中有机氯农药的残留分析。
Quality is one of the critical factors that Chinese materia medica enter into the international market,and the quality determination of Chinese materia medica is an important process of their admission to China market,both of which bring forward some high requirement to the quality control of Chinese materia medica.It is a feasible approach that applying chromatography to analyzing and testing not only effective components,but also harmful components in Chinese materia medica for quality control.
Referred to a lot of literatures,this paper improved the quality control method of the Composite Salvia Pellet and Gujing Pills.The main contains three parts.First.the effective component borneol in the Composite Salvia Pellet,has been analyzed by gas chromatography and high performance liquid chromatography.Borneol has volatility.So it is a mature method to determine the content of borneol by gas chromatography.But because the UV absorption spectra of borneol has no Characteristics,so it is difficult to determine it by high performance liquid chromatography directly.According to the feature of hydroxy in the borneol,it could be determined by HPLC derivatization method. By pre-column derivatization with 3,5-dinitrobenzoyl chloride,catalyzed by tertiary amine or pyridines,the intensive-UV-absorbed borneyl 3,5-dinitrobenzoate was generated.So the borneol in the Composite Salvia Pellet was determined by the two methods to validate the accuracy of the latter.The average recovery of borneol is 101.2% and RSD is 3.1%.The testing solution for assay is stable within 8 hours.The HPLC experimental results indicate that chromatographic peak areas are linear with quantity of injecting sample in the range of 1～10nmol for borneol.The average recovery rate is 99.8%and RSD is 1.6%(n=4).It can be found from contrasting that the measured contents in the two methods are equivalent,so they both can be used for the determination of borneol content.
And then,the content of Paeoniflorin in Gujing Pills has been determined by applying RP-HPLC to improve the quality control method of this Chinese materia medica.A good linear relationship is in the range of 0.44～3.96μg(r=0.9999),the average recovery is 99.2%(n=5),and the repetitive RSD is 0.82%(n=5).The testing solution for assay is stable within 24 hours.The results show that HPLC can be used in the quality control of Chinese materia medica containing the effective component paeoniflorin.
Finally,the organochlorine pesticide in Composite Salvia Pellet is determined by applying capillary gas chromatography.The experimental results show that a good linear relationship of the 11 organochlorine pesticides is in the range of(1.0×10~(-4)～5.0×10~(-1))μg/ml,and the correlation coefficient is in the range of 0.9988-0.9999.The average recovery is in the range of 74.2～101.5%(n=6),and RSD is in the range of 2.4～8.7%. This method can be used for the determination of organochlorine pesticide content in Chinese materia medica.
本论文在查阅大量文献的基础上,对复方丹参滴丸和固经丸的质量控制方法进行改进。主要内容包括三个部分。首先采用毛细管气相色谱法及反相高效液相色谱法分别测定了复方丹参滴丸中冰片的含量。冰片具有挥发性,因此气相色谱法测冰片含量的方法较为成熟。但是由于冰片无特征紫外吸收,所以直接采用高效液相色谱检测比较困难。根据冰片结构中含羟基的特点,可采用高效液相色谱衍生法检测。3,5-二硝基苯甲酰氯作为柱前衍生化试剂,在叔胺或吡啶类化合物的催化下可反应生成具强紫外吸收的3,5-二硝基苯甲酸冰片酯。故将两种方法分别用于测定复方丹参滴丸中冰片含量,以验证后者的准确性。冰片的平均回收率为101.2%,RSD为3.1%。供试品溶液在8h内稳定。高效液相色谱法测定的实验结果表明,在1～10nmol范围内,色谱峰面积与进样量呈线性关系。复方丹参滴丸中冰片的回收率为99.8%,RSD为1.6%(n=4)。对比发现,两种方法所测含量相当,均可用于测定冰片含量。
其次,采用反相高效液相色谱法测定了固经丸中有效成分芍药苷的含量,改进了该中药制剂的质量控制方法。芍药苷在0.44～3.96μg范围内的线性关系良好(r=0.9999),平均加样回收率为99.2%(n=5),重现性试验RSD为0.82%(n=5)。供试品溶液在24h内稳定性良好。结果表明高效液相色谱法可用于含有效成分芍药苷的中药制剂的质量控制。
最后以毛细管气相色谱法测定了复方丹参滴丸中有机氯农药的残留量。实验结果表明,11种农药在1.0×10~(-4)～5.0×10~(-1)μg/ml范围内的线性关系良好,相关系数在0.9988～0.9999范围之间。平均回收率在74.2～101.5%(n=6)范围内,RSD在2.4～8.7%之间,该方法可用于中药制剂中有机氯农药的残留分析。
Quality is one of the critical factors that Chinese materia medica enter into the international market,and the quality determination of Chinese materia medica is an important process of their admission to China market,both of which bring forward some high requirement to the quality control of Chinese materia medica.It is a feasible approach that applying chromatography to analyzing and testing not only effective components,but also harmful components in Chinese materia medica for quality control.
Referred to a lot of literatures,this paper improved the quality control method of the Composite Salvia Pellet and Gujing Pills.The main contains three parts.First.the effective component borneol in the Composite Salvia Pellet,has been analyzed by gas chromatography and high performance liquid chromatography.Borneol has volatility.So it is a mature method to determine the content of borneol by gas chromatography.But because the UV absorption spectra of borneol has no Characteristics,so it is difficult to determine it by high performance liquid chromatography directly.According to the feature of hydroxy in the borneol,it could be determined by HPLC derivatization method. By pre-column derivatization with 3,5-dinitrobenzoyl chloride,catalyzed by tertiary amine or pyridines,the intensive-UV-absorbed borneyl 3,5-dinitrobenzoate was generated.So the borneol in the Composite Salvia Pellet was determined by the two methods to validate the accuracy of the latter.The average recovery of borneol is 101.2% and RSD is 3.1%.The testing solution for assay is stable within 8 hours.The HPLC experimental results indicate that chromatographic peak areas are linear with quantity of injecting sample in the range of 1～10nmol for borneol.The average recovery rate is 99.8%and RSD is 1.6%(n=4).It can be found from contrasting that the measured contents in the two methods are equivalent,so they both can be used for the determination of borneol content.
And then,the content of Paeoniflorin in Gujing Pills has been determined by applying RP-HPLC to improve the quality control method of this Chinese materia medica.A good linear relationship is in the range of 0.44～3.96μg(r=0.9999),the average recovery is 99.2%(n=5),and the repetitive RSD is 0.82%(n=5).The testing solution for assay is stable within 24 hours.The results show that HPLC can be used in the quality control of Chinese materia medica containing the effective component paeoniflorin.
Finally,the organochlorine pesticide in Composite Salvia Pellet is determined by applying capillary gas chromatography.The experimental results show that a good linear relationship of the 11 organochlorine pesticides is in the range of(1.0×10~(-4)～5.0×10~(-1))μg/ml,and the correlation coefficient is in the range of 0.9988-0.9999.The average recovery is in the range of 74.2～101.5%(n=6),and RSD is in the range of 2.4～8.7%. This method can be used for the determination of organochlorine pesticide content in Chinese materia medica.
引文
[1]刘文英.药物分析,人民卫生出版社,2006,第5版,347-370.
[2]潘旭初,吴继洲.时珍国药研究.1998,9(1):5.
[3]贺贤国,蒋福祥,周倩如.植物学报,1978,20(1):77.
[4]张仁斌.高效液相色谱在医药研究中的应用.上海科技出版社,1983,第1版.
[5]杨军.时珍国药研究,1998,9(2):126.
[6]赵芝兰.药学实践杂志,1998,16(2):97.
[7]李华斌.中国卫生检验杂志,1998,8(2):123.
[8]刘虎威.气相色谱方法及应用.化学工业出版社,2007,第2版,99-120.
[9]张志兰等.中国民族医药杂志,2003,9(1):38.
[10]王凌,季申.中草药,2003,34(3):226.
[11]邵彩云,刘永喜.中国民族医药杂志,2003,9(1):37.
[12]周祥敏,谢静.中国药业,2004,13(6):46.
[13]聂少平,王远兴,谢明勇.中成药,2004,16(1):80.
[14]解荷芝,何志强,陈吴苏等.中医药学刊,2004,22(8):1560.
[15]魏学冰.中成药.2004,26(1):76
[16]杨立新,崔淑莲,刘岱等.中国实验方剂学杂志,2003,9(6):5.
[17]刘杰,张正行,都恒青.中医研究,2004,17(1):19.
[18]周利贤,吴陈军.中国药师.2004,7(6):430-431.
[19]刘振,潘诗海,段更利.中国临床药学杂志.2004,13(1):32.
[20]田薇,陈朝晖,柳春晖等.中成药.2004,26(8):624.
[21]周静,刘垣升.中国新药民临床杂志.2003,22(10):596.
[22]崔莉凤,王微.色谱.2003,21(1):88.
[23]丁旭光,张捷莉,郑杰等.时珍国医国药.2005,16(3):202.
[24]Ling Dong,Chunhui Deng,Jiyao Wang,et al.Anal ChimActa.2007,585(1):76.
[25]Ling Dong,Jiyao Wang,Chunhui Deng,et al.J Sep Sci.2007,30(1):86.
[26]田金改,高天兵,张曙明等.中国中药杂志,2000,25(5):279-282.
[27]何笑荣,邹定,姜文清等.中国药学杂志,2006,41(9):704-706.
[28]郭素华,廖华军.中成药.2005,27(2):1-164.
[29]王智华,朱静建,洪筱坤.中成药.1994,16(7):42-44.
[30]汪锡灿,周素娣,盛娟芬等.中国药科大学学报,1996,27(1):32-35.
[31]田金改,金红宇,杜庆鹏等.中国药事,2006,20(12):755-758.
[32]Zuin V.G..,Yariwake J.H.,Bicchi C..J.Chromatogr.A,2003,985:159.
[33]Hwang B.H.,Lee M.R..J.Chromatogr.A,2000,898(2):245.
[34]Barriada-Perira M.,Concha-Grana.E,Gonzalez-Castro MJ.J.Chromatogr.A,2003,1008(1):115-122.
[35]Ling,Y.C.,Teng,H.C.,Cartwright.C..J.Chromatogr.A,1999,835(1-2):145-157.
[36]C.Goncalves,M.F.Alpendurada.J.Chromatogr.A,2004,1026:239-250.
[37]L.Cai,et al.J.Chromatogr.A,2003,1015:11-21.
[38]F.Smedes,J.de Boer.Trends Anal.Chem.,1997,16:503.
[39]Vanderhoff G.R.,Vanzoonen P..J.Chromatogr.A.1999,843:301-322.
[40]Torres C.M.,Pico Y.,Manes J..J.Chromatogr.A,1996,754:301-331.
[41]Kim J.,Smith A..J.Chromatogr.A,1996,725:93-106.
[42]Norman K.N.T.,Panton S.H.W..J.Chromatogr.A,2001,907:247-255.
[43]Sakamoto M.,Tsutsumi T..J.Chromatogr.A,2000,885:115-127.
[44]Navarro M.,Pico Y.,Marn R.,et al.J.Chromatogr.A,2002,982:267-273.
[45]中国药典委员会.中国药典[S].2005年版.一部.528
[46]丁宁.中草药.2002,33(12):1147-1148.
[47]向柏,潘振华,曹德英.中国药房,2007,18(9):708-710.
[48]刘景英.天津医科大学学报,2004,10(2):195-196.
[49]靳凤云,赵杨,廖薇等.World Health Digest.2007,4(11):49-50.
[50]梁燕,李蓉.时珍国医国药.2007,18(5):1132-1133.
[1]李伟佳,王强.中成药,2007,29(8):1177-1179.
[2]张韵,张美怡,李晓蒙.中药材,2007,30(8):1033-1035.
[3]梁骏,陈华.中药材,2007,30(4):483-484.
[4]王苑,曾荣仕,朱湛华.中国药房,2007,18(27):2124-2126.
[5]王姝,车晶玉,卢盛贞等.辽宁中医杂志,2003,30(7):581.
[1]杨丽华.中国中药杂志,2000,25(2):122-124.
[2]赵永德,丁昌成,贾善学.中国药师,2005,8(12):1007-1008.
[3]中国药典委员会.中国药典[S].2005年版.一部.494.
[1]张亚莉,习立鹏,阎正.河北科技大学学报.2005,26(4):275-276,333.
[2]赵春杰,郝桂明,李欢欣.中国药学杂志,2002,37(7):47-50.
[3]宋家玉,李泽国,王大卫等.化学分析计量,2006,15(6):8-10
[4]李庆,邹巧根.中草药,2001,32(5):33-34.
[5]杨玉琴,张丽艳,蒋朝辉.中药材.2002,25(9):625-626.
[6]田金改,高天兵,张曙明等.中国中药杂志,2000,25(5):279-282.
[7]何笑荣,邹定,姜文清等.中国药学杂志,2006,41(9):704-706.
[8]中国药典委员会.中国药典[S].2005年版.一部.附录52-53.
[2]潘旭初,吴继洲.时珍国药研究.1998,9(1):5.
[3]贺贤国,蒋福祥,周倩如.植物学报,1978,20(1):77.
[4]张仁斌.高效液相色谱在医药研究中的应用.上海科技出版社,1983,第1版.
[5]杨军.时珍国药研究,1998,9(2):126.
[6]赵芝兰.药学实践杂志,1998,16(2):97.
[7]李华斌.中国卫生检验杂志,1998,8(2):123.
[8]刘虎威.气相色谱方法及应用.化学工业出版社,2007,第2版,99-120.
[9]张志兰等.中国民族医药杂志,2003,9(1):38.
[10]王凌,季申.中草药,2003,34(3):226.
[11]邵彩云,刘永喜.中国民族医药杂志,2003,9(1):37.
[12]周祥敏,谢静.中国药业,2004,13(6):46.
[13]聂少平,王远兴,谢明勇.中成药,2004,16(1):80.
[14]解荷芝,何志强,陈吴苏等.中医药学刊,2004,22(8):1560.
[15]魏学冰.中成药.2004,26(1):76
[16]杨立新,崔淑莲,刘岱等.中国实验方剂学杂志,2003,9(6):5.
[17]刘杰,张正行,都恒青.中医研究,2004,17(1):19.
[18]周利贤,吴陈军.中国药师.2004,7(6):430-431.
[19]刘振,潘诗海,段更利.中国临床药学杂志.2004,13(1):32.
[20]田薇,陈朝晖,柳春晖等.中成药.2004,26(8):624.
[21]周静,刘垣升.中国新药民临床杂志.2003,22(10):596.
[22]崔莉凤,王微.色谱.2003,21(1):88.
[23]丁旭光,张捷莉,郑杰等.时珍国医国药.2005,16(3):202.
[24]Ling Dong,Chunhui Deng,Jiyao Wang,et al.Anal ChimActa.2007,585(1):76.
[25]Ling Dong,Jiyao Wang,Chunhui Deng,et al.J Sep Sci.2007,30(1):86.
[26]田金改,高天兵,张曙明等.中国中药杂志,2000,25(5):279-282.
[27]何笑荣,邹定,姜文清等.中国药学杂志,2006,41(9):704-706.
[28]郭素华,廖华军.中成药.2005,27(2):1-164.
[29]王智华,朱静建,洪筱坤.中成药.1994,16(7):42-44.
[30]汪锡灿,周素娣,盛娟芬等.中国药科大学学报,1996,27(1):32-35.
[31]田金改,金红宇,杜庆鹏等.中国药事,2006,20(12):755-758.
[32]Zuin V.G..,Yariwake J.H.,Bicchi C..J.Chromatogr.A,2003,985:159.
[33]Hwang B.H.,Lee M.R..J.Chromatogr.A,2000,898(2):245.
[34]Barriada-Perira M.,Concha-Grana.E,Gonzalez-Castro MJ.J.Chromatogr.A,2003,1008(1):115-122.
[35]Ling,Y.C.,Teng,H.C.,Cartwright.C..J.Chromatogr.A,1999,835(1-2):145-157.
[36]C.Goncalves,M.F.Alpendurada.J.Chromatogr.A,2004,1026:239-250.
[37]L.Cai,et al.J.Chromatogr.A,2003,1015:11-21.
[38]F.Smedes,J.de Boer.Trends Anal.Chem.,1997,16:503.
[39]Vanderhoff G.R.,Vanzoonen P..J.Chromatogr.A.1999,843:301-322.
[40]Torres C.M.,Pico Y.,Manes J..J.Chromatogr.A,1996,754:301-331.
[41]Kim J.,Smith A..J.Chromatogr.A,1996,725:93-106.
[42]Norman K.N.T.,Panton S.H.W..J.Chromatogr.A,2001,907:247-255.
[43]Sakamoto M.,Tsutsumi T..J.Chromatogr.A,2000,885:115-127.
[44]Navarro M.,Pico Y.,Marn R.,et al.J.Chromatogr.A,2002,982:267-273.
[45]中国药典委员会.中国药典[S].2005年版.一部.528
[46]丁宁.中草药.2002,33(12):1147-1148.
[47]向柏,潘振华,曹德英.中国药房,2007,18(9):708-710.
[48]刘景英.天津医科大学学报,2004,10(2):195-196.
[49]靳凤云,赵杨,廖薇等.World Health Digest.2007,4(11):49-50.
[50]梁燕,李蓉.时珍国医国药.2007,18(5):1132-1133.
[1]李伟佳,王强.中成药,2007,29(8):1177-1179.
[2]张韵,张美怡,李晓蒙.中药材,2007,30(8):1033-1035.
[3]梁骏,陈华.中药材,2007,30(4):483-484.
[4]王苑,曾荣仕,朱湛华.中国药房,2007,18(27):2124-2126.
[5]王姝,车晶玉,卢盛贞等.辽宁中医杂志,2003,30(7):581.
[1]杨丽华.中国中药杂志,2000,25(2):122-124.
[2]赵永德,丁昌成,贾善学.中国药师,2005,8(12):1007-1008.
[3]中国药典委员会.中国药典[S].2005年版.一部.494.
[1]张亚莉,习立鹏,阎正.河北科技大学学报.2005,26(4):275-276,333.
[2]赵春杰,郝桂明,李欢欣.中国药学杂志,2002,37(7):47-50.
[3]宋家玉,李泽国,王大卫等.化学分析计量,2006,15(6):8-10
[4]李庆,邹巧根.中草药,2001,32(5):33-34.
[5]杨玉琴,张丽艳,蒋朝辉.中药材.2002,25(9):625-626.
[6]田金改,高天兵,张曙明等.中国中药杂志,2000,25(5):279-282.
[7]何笑荣,邹定,姜文清等.中国药学杂志,2006,41(9):704-706.
[8]中国药典委员会.中国药典[S].2005年版.一部.附录52-53.