小檗皮总生物碱提取物的大孔树脂纯化工艺与质量标准考察
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摘要
目的:优选小檗皮总生物碱提取物的大孔树脂纯化工艺,并建立其质量标准,为该有效部位的制剂开发提供参考。方法:采用酸性染料比色法考察小檗皮总生物碱提取物的纯化工艺,考察的工艺参数包括上样液质量浓度、上样速度、树脂柱径高比、水洗用量、洗脱剂体积分数及用量、洗脱流速等。采用HPLC测定小檗皮总生物碱提取物中4种生物碱类成分(木兰花碱、盐酸药根碱、盐酸巴马汀、盐酸小檗碱)的含量,流动相乙腈-0. 1%磷酸水溶液梯度洗脱,检测波长270 nm,确定最佳纯化工艺。按2015年版《中国药典》的要求进行小檗皮总生物碱纯化物的薄层色谱鉴别、含量测定、特征图谱等质量标准研究。结果:小檗皮总生物碱提取物的最佳纯化条件为采用HPD100型大孔树脂10 g,树脂柱径高比1∶8,上样液质量浓度11 g·L~(-1),上样液体积50 mL,上样流速1 mL·min~(-1),加4 BV水洗脱(1 BV=15 mL),加30%乙醇9 BV洗脱;纯化后小檗皮总生物碱的转移率> 80%,纯度> 65%。建立了小檗皮总生物碱纯化物的质量标准,特征图谱中共有峰有19个,整体相似度均> 0. 99。结论:优选的纯化工艺稳定可行,建立的质量标准可靠,适用于小檗皮总生物碱提取物的纯化及质量控制。
Objective: To optimize purification process of total alkaloid extract of Berberis dictyophylla cortex by macroporous resin,and to establish its quality standard. Method: Acid dye colorimetry was used to investigate the purification process of total alkaloid extract of B. dictyophylla cortex, the process parameters included concentration of sample solution, speed of sampling, diameter-height ratio of resin column, water washing amount,concentration and dosage of eluent,flow rate of elution,etc. In order to determine the optimum process, HPLC was employed to determine the contents of four alkaloids( magnoflorine, jatrorrhizine hydrochloride,palmatine hydrochloride,and berberine hydrochloride) with mobile phase of acetonitrile -0. 1%phosphoric acid aqueous solution for gradient elution and detection wavelength at 270 nm. After being purified,quality standard of total alkaloid extract of B. dictyophylla cortex was investigated according to the requirements in the 2015 edition of Chinese Pharmacopoeia. Result: Optimal purification conditions were as following: 10 g of HPD100 macroporous adsorption resin with a column diameter-height ratio of 1 ∶ 8,sampling solution concentration of 11 g·L~(-1),the loading flow rate of 1 mL·min~(-1),sampling solution volume of 50 mL,washed with 4 BV of water( 1 BV = 15 mL) and added 9 BV of 30% ethanol,after being purified,the transfer rate of total alkaloids was > 80%,and its purity was > 65%. The quality standard of total alkaloid extract of B. dictyophylla cortex was established,there were 19 common peaks in the characteristic chromatogram,and the overall similarity was >0. 99. Conclusion: This optimized purification process is stable and feasible,and the established quality standard is controllable.
Objective: To optimize purification process of total alkaloid extract of Berberis dictyophylla cortex by macroporous resin,and to establish its quality standard. Method: Acid dye colorimetry was used to investigate the purification process of total alkaloid extract of B. dictyophylla cortex, the process parameters included concentration of sample solution, speed of sampling, diameter-height ratio of resin column, water washing amount,concentration and dosage of eluent,flow rate of elution,etc. In order to determine the optimum process, HPLC was employed to determine the contents of four alkaloids( magnoflorine, jatrorrhizine hydrochloride,palmatine hydrochloride,and berberine hydrochloride) with mobile phase of acetonitrile -0. 1%phosphoric acid aqueous solution for gradient elution and detection wavelength at 270 nm. After being purified,quality standard of total alkaloid extract of B. dictyophylla cortex was investigated according to the requirements in the 2015 edition of Chinese Pharmacopoeia. Result: Optimal purification conditions were as following: 10 g of HPD100 macroporous adsorption resin with a column diameter-height ratio of 1 ∶ 8,sampling solution concentration of 11 g·L~(-1),the loading flow rate of 1 mL·min~(-1),sampling solution volume of 50 mL,washed with 4 BV of water( 1 BV = 15 mL) and added 9 BV of 30% ethanol,after being purified,the transfer rate of total alkaloids was > 80%,and its purity was > 65%. The quality standard of total alkaloid extract of B. dictyophylla cortex was established,there were 19 common peaks in the characteristic chromatogram,and the overall similarity was >0. 99. Conclusion: This optimized purification process is stable and feasible,and the established quality standard is controllable.
引文
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[10]吴秦西,王友利,张燕,等.均匀设计法优化藏药小檗皮的煎煮法提取工艺[J].时珍国医国药,2013,24(7):1637-1639.
[11]郝露,周珍,冯慧,等.藏药小檗皮的醇提工艺优化研究[J].中国药房,2018,29(7):958-963.
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[15]李路扬,龙娓芳,万定荣,等. 5种小檗属药用植物总生物碱及小檗碱的含量测定[J].中国实验方剂学杂志,2014,20(17):91-94.
[16]费艳美,张玉萍,方步武,等.白英中总生物碱的含量测定[J].西北药学杂志,2009,24(4):260-262.
[17]刘喜纲,刘翠哲,常金花.应用酸性染料比色法测定总生物碱的含量[J].中国药房,2007,18(11):875-876.
[18]甄会贤,李平,解育静,等.酸性染料比色法测定延胡索总生物碱的含量[J].现代中药研究与实践,2012,26(3):66-68.
[19]范圣洁,李文龙,崔文霞,等.酸性染料比色法测定飞龙掌血提取物中总生物碱的含量[J].上海中医药大学学报,2012,26(1):85-89.
[20]张旭,王锦玉,仝燕,等.大孔树脂技术在中药提取纯化中的应用及展望[J].中国实验方剂学杂志,2012,18(6):286-290.
[21]王玉霞,刘斌,姜艳艳.荷叶生物碱提取纯化工艺研究[J].北京中医药大学学报,2013,36(9):622-626.
[22]王毓杰.管花肉苁蓉中苯乙醇苷的提取和大孔树脂纯化工艺研究[J].中草药,2014,45(16):2344-2348.
[23]孟甜.红景天总鞣质的提取纯化工艺及质量标准研究[D].成都:西华大学,2010.
[24]国家药典委员会.中华人民共和国药典.四部[M].北京:中国医药科技出版社,2015:57-59,101-104,204.
[25]刘芳,张浩,青琳森.黄连HPLC数字化指纹图谱研究及7种生物碱含量测定[J].中国中药杂志,2013,38(21):3713-3719.
[26]谢巍,杨妮,蒋受军.白背叶HPLC特征指纹图谱及模式识别[J].中国实验方剂学杂志,2017,23(18):52-57.
[27]余佳丽,唐晓章,周菲,等.三七药材-三七二醇皂苷指纹图谱及其物质群量值传递分析[J].中国实验方剂学杂志,2018,24(7):18-22.
[2]西藏、青海、四川、甘肃、云南、新疆卫生局.藏药标准(第一、二分册合编本)[M].西宁:青海人民出版社,1979:11.
[3]青海省药品检验所,青海省藏医药研究所.中国藏药.第三卷[M].上海:上海科学技术出版社,1996:28-30.
[4]罗达尚.中华藏本草[M].北京:民族出版社,1997:84-85.
[5]吕秀梅,李艳,莫家祺,等.藏族药小檗皮不同药用部位中4种生物碱成分的HPLC含量测定[J].中国实验方剂学杂志,2017,23(2):37-42.
[6]岳丽珺.藏药吉尔巴对自发性糖尿病db/db小鼠视网膜病变的药效及机制研究[D].成都:成都中医药大学,2014.
[7]张燕,孟宪丽,岳丽珺,等.藏药小檗皮对糖尿病模型小鼠血糖水平影响的初步研究[J].现代生物医学进展,2013,13(19):3619-3622.
[8] Pirillo A,Catapano A L. Berberine,a plant alkaloid with lipid-and glucose-lowering properties:from in vitro evidence to clinical studies[J]. Atherosclerosis,2015,243(2):449-461.
[9]叶凡,周邦华,王胜,等.藏药小檗膏提取物中3种生物碱含量的高效液相色谱法测定[J].时珍国医国药,2016,27(2):286-289.
[10]吴秦西,王友利,张燕,等.均匀设计法优化藏药小檗皮的煎煮法提取工艺[J].时珍国医国药,2013,24(7):1637-1639.
[11]郝露,周珍,冯慧,等.藏药小檗皮的醇提工艺优化研究[J].中国药房,2018,29(7):958-963.
[12]叶凡.藏药小檗皮提取物的成药性研究[D].成都:成都中医药大学,2016.
[13]张林,张朝凤,许翔鸿,等.生物碱提取分离方法研究进展[J].亚太传统医药,2017,13(15):49-50.
[14]匡海学.中药化学[M].北京:中国中医药出版社,2005:313-318.
[15]李路扬,龙娓芳,万定荣,等. 5种小檗属药用植物总生物碱及小檗碱的含量测定[J].中国实验方剂学杂志,2014,20(17):91-94.
[16]费艳美,张玉萍,方步武,等.白英中总生物碱的含量测定[J].西北药学杂志,2009,24(4):260-262.
[17]刘喜纲,刘翠哲,常金花.应用酸性染料比色法测定总生物碱的含量[J].中国药房,2007,18(11):875-876.
[18]甄会贤,李平,解育静,等.酸性染料比色法测定延胡索总生物碱的含量[J].现代中药研究与实践,2012,26(3):66-68.
[19]范圣洁,李文龙,崔文霞,等.酸性染料比色法测定飞龙掌血提取物中总生物碱的含量[J].上海中医药大学学报,2012,26(1):85-89.
[20]张旭,王锦玉,仝燕,等.大孔树脂技术在中药提取纯化中的应用及展望[J].中国实验方剂学杂志,2012,18(6):286-290.
[21]王玉霞,刘斌,姜艳艳.荷叶生物碱提取纯化工艺研究[J].北京中医药大学学报,2013,36(9):622-626.
[22]王毓杰.管花肉苁蓉中苯乙醇苷的提取和大孔树脂纯化工艺研究[J].中草药,2014,45(16):2344-2348.
[23]孟甜.红景天总鞣质的提取纯化工艺及质量标准研究[D].成都:西华大学,2010.
[24]国家药典委员会.中华人民共和国药典.四部[M].北京:中国医药科技出版社,2015:57-59,101-104,204.
[25]刘芳,张浩,青琳森.黄连HPLC数字化指纹图谱研究及7种生物碱含量测定[J].中国中药杂志,2013,38(21):3713-3719.
[26]谢巍,杨妮,蒋受军.白背叶HPLC特征指纹图谱及模式识别[J].中国实验方剂学杂志,2017,23(18):52-57.
[27]余佳丽,唐晓章,周菲,等.三七药材-三七二醇皂苷指纹图谱及其物质群量值传递分析[J].中国实验方剂学杂志,2018,24(7):18-22.
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