大孔吸附树脂纯化延胡索总生物碱工艺研究
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摘要
目的确定大孔吸附树脂纯化延胡索总生物碱的工艺条件。方法以酸性染料比色法测定延胡索总生物碱,HPLC法测定延胡索中盐酸巴马汀、脱氢紫堇碱、延胡索乙素及延胡索甲素;以延胡索总生物碱和4种生物碱的吸附率、洗脱率及质量分数为指标,利用静态吸附和动态吸附实验相结合的方法,对6种大孔吸附树脂进行筛选。通过考察上样量、体积流量、除杂溶剂和体积、洗脱溶剂和体积等因素,优选延胡索总生物碱纯化工艺条件,以5、10倍放大试验考察纯化工艺的稳定性。结果选取D141型大孔树脂用于延胡索总生物碱纯化,纯化工艺为生药0.6 g/m L的上样液,加入径高比1∶5~1∶9的D141大孔吸附树脂柱,以体积流量2 BV/h上样2 BV,1.3 BV纯水除杂,6 BV 95%乙醇以2 BV/h体积流量洗脱。纯化后延胡索总生物碱质量分数高达68.19%,盐酸巴马汀、脱氢紫堇碱、延胡索乙素及延胡索甲素的质量分数分别为1.95%、11.74%、4.93%、6.36%。经5、10倍放大试验得到的延胡索总生物碱质量分数能达到65%以上。结论该工艺经验证试验,总生物碱质量分数能达到65%以上,且延胡索总生物碱及4种生物碱单体的转移率均能达到85%以上,表明D141型大孔吸附树脂能有效纯化延胡索中总生物碱,且适用于工业大生产。
Objective To determine the technological conditions for the purification of the total alkaloid from the Corydalis Rhizoma(CR) by macroporous adsorption resin. Methods Total alkaloids of CR were determined by acid dye colorimetry, palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline were determined by HPLC. Six macroporous adsorption resins were investigated with the absorption rates, elution rates, and the content of the total alkaloid and four alkaloids of CR by static and dynamic adsorbing experiments. The purification process conditions of the total alkaloid of CR were optimized by the loading amount and volume flow of sample, the type and volume of the impurity removal of solvent and elution solvent and so on. The stability of the purification process was investigated by 5, 10 times enlargement. Results D141 type macroporous adsorption resin was the best choice for the purification of the total alkaloid from CR, the optimized parameters were as follows: Drug concentration was 0.6 g/mL of medicinal material and was added to the D141 macroporous resin column that the ratio of diameter to height range from 1∶5 to 1∶9 at a flow rate of 2 BV/h to 2 BV, 1.3 BV of purified water was used to remove impurities, and then 6 BV 95% ethanol was used as eluent at a flow rate of 2 BV/h. The purity of the total alkaloid of CR was up to 68.19% after purification, and the content of palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline was 1.95%, 11.74%, 4.93%, and 6.36%, respectively. The purity of the total alkaloid of the CR can reach more than 65% by 5 times and 10 times enlargement. Conclusion The purity of the total alkaloids can reach more than 65% after verification test, and the transfer rate of total alkaloids and four alkaloid monomers of CR can reach more than 85%, indicating that D141 macroporous adsorption resin can effectively purify total alkaloids from CR, and can be applied to industrial production.
Objective To determine the technological conditions for the purification of the total alkaloid from the Corydalis Rhizoma(CR) by macroporous adsorption resin. Methods Total alkaloids of CR were determined by acid dye colorimetry, palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline were determined by HPLC. Six macroporous adsorption resins were investigated with the absorption rates, elution rates, and the content of the total alkaloid and four alkaloids of CR by static and dynamic adsorbing experiments. The purification process conditions of the total alkaloid of CR were optimized by the loading amount and volume flow of sample, the type and volume of the impurity removal of solvent and elution solvent and so on. The stability of the purification process was investigated by 5, 10 times enlargement. Results D141 type macroporous adsorption resin was the best choice for the purification of the total alkaloid from CR, the optimized parameters were as follows: Drug concentration was 0.6 g/mL of medicinal material and was added to the D141 macroporous resin column that the ratio of diameter to height range from 1∶5 to 1∶9 at a flow rate of 2 BV/h to 2 BV, 1.3 BV of purified water was used to remove impurities, and then 6 BV 95% ethanol was used as eluent at a flow rate of 2 BV/h. The purity of the total alkaloid of CR was up to 68.19% after purification, and the content of palmatine hydrochloride, dehydrocorydaline, tetrahydropalmatine, and corydaline was 1.95%, 11.74%, 4.93%, and 6.36%, respectively. The purity of the total alkaloid of the CR can reach more than 65% by 5 times and 10 times enlargement. Conclusion The purity of the total alkaloids can reach more than 65% after verification test, and the transfer rate of total alkaloids and four alkaloid monomers of CR can reach more than 85%, indicating that D141 macroporous adsorption resin can effectively purify total alkaloids from CR, and can be applied to industrial production.
引文
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[20]徐丽琳,熊雪丰,何三民,等.元胡总生物碱的大孔树脂纯化工艺研究[J].中华中医药学刊,2013,31(6):1358-1361.
[21]蒲忠慧,王力,高宇,等.大孔吸附树脂纯化富集川芎总生物碱的工艺[J].食品研究与开发,2017,38(11):85-91.
[22]季慧,陈斌斌,黄越燕.大孔吸附树脂分离纯化生物碱类化合物研究进展[J].亚太传统医药,2017,13(1):72-75.
[2]王桂彩,应森林.重用延胡索(元首汤、二胡汤)从肝论治顽固性失眠[J].实用中医内科杂志,2015,29(8):57-58.
[3]赵丽沙,董宇,寿旦.延胡索生物碱类化学成分及质量控制研究进展[J].中华中医药学刊,2017,35(2):299-302.
[4]王磊,王媛,李爱主,等.基于靶细胞萃取的延胡索药效成分的筛选和含量测定[J].药物评价研究,2016,39(6):990-994.
[5]杜巍娟.延胡索提取物在小鼠体内的药代动力学研究[D].杭州:浙江大学,2017.
[6]李秋月.延胡索生物碱谱-效关系及相互作用研究[D].北京:北京协和医学院,中国医学科学院,清华大学医学部,2014.
[7]张晓峰,张宏,李小云,等.高速逆流色谱-UPLCQ-TOF-MS/MS法分离制备延胡索中脱氢紫堇碱和海罂粟碱[J].中草药,2016,47(24):4351-4356.
[8]施婷婷,王建新,李希.延胡索总生物碱类有效部位的研究进展[J].中药与临床,2015,6(2):110-113.
[9]郭倩,田成旺,任涛,等.中药药效物质基础研究进展[J].世界科学技术—中医药现代化,2015,17(3):648-654.
[10]汤如莹,王玉杰,李伟,等.D101型大孔吸附树脂分离纯化沙苑子总黄酮工艺研究[J].中草药,2017,48(16):3342-3346.
[11]任慧梅,盛萍.大孔吸附树脂纯化富集伊贝母总生物碱的工艺研究[J].中国现代应用药学,2016,33(6):695-699.
[12]李士杰,肖连冬.水提醇沉法提取苦瓜多糖条件研究[J].中国酿造,2016,35(5):166-170.
[13]刘丹,吴叶红,李玮桓,等.大孔吸附树脂在天然产物分离纯化中的应用[J].中草药,2016,47(15):2764-2770.
[14]Zhu S,Bo T,Wang X,et al.Separation of succinic acid from aqueous solution by macroporous resin adsorption[J].J Chem Eng Data,2016,61(2):856-864.
[15]Zhao Y,Chen Z,Liu D,et al.Separation of flavonoids in the leaves of Sophora japonica by macroporous adsorption resin mixed-bed technology[J].Pigm ResinTechnol,2017,46(3):235-243.
[16]Liu C,Liu R,Zhang P,et al.Separation of capsaicin from capsaicinoids by macroporous resin adsorption chromatography[J].J Sep Sci,2015,38(23):4141-4145.
[17]韩璐,杨杰,王丽莉,等.大孔吸附树脂纯化黄花草木犀的工艺研究[J].现代药物与临床,2015,30(7):800-804.
[18]吕子明,刘文娟,于向红,等.大孔吸附树脂纯化延胡索总生物碱工艺研究[J].中国医药导报,2012,20(9):109-112.
[19]罗雪芹,何道惠,秦秀蓉,等.延胡索有效成分提取纯化的工艺研究[J].时珍国医国药,2014,25(1):23-25.
[20]徐丽琳,熊雪丰,何三民,等.元胡总生物碱的大孔树脂纯化工艺研究[J].中华中医药学刊,2013,31(6):1358-1361.
[21]蒲忠慧,王力,高宇,等.大孔吸附树脂纯化富集川芎总生物碱的工艺[J].食品研究与开发,2017,38(11):85-91.
[22]季慧,陈斌斌,黄越燕.大孔吸附树脂分离纯化生物碱类化合物研究进展[J].亚太传统医药,2017,13(1):72-75.