HPLC Determination of 5 Alkaloids in Sophora Flavescens Ait with Microwave Assisted Extraction
摘 要
取粒径在0.18~0.25 mm的药材粉末于提取罐中,用水作提取剂,在110 ℃、压力为15 MPa、功率为780 W的条件下进行微波提取(PMAE) 5 min,经离心后取上清液为提取液。提取液经无水乙醇处理提纯后得到待测物的甲醇溶液,从中分取部分溶液按选定条件用液相色谱-质谱(LC-MS)联用做定性分析,用高效液相色谱(HPLC)做定量分析。LC-MS结果表明:提取液中含有氧化苦参碱、氧化槐果碱、槐定碱、苦参碱和槐果碱,与其它3种提取方法比较,PMAE的提取率高且提取时间短。HPLC定量时,采用C18色谱柱,以不同比例的甲醇、乙腈和0.1%氨水溶液(pH 10.3)的混合溶液作流动相进行梯度淋洗,可分离上述5种生物碱,以峰面积和浓度间的线性关系进行定量。5种生物碱的回收率在93.9%~101.0%之间,测定值的相对标准偏差(n=5)在0.53%~2.6%之间。
Abstract
Sample of the crushed drug with sizes between 0.18-0.25 mm was placed in a closed extraction vessel and treated by microwave assisted extraction for 5 min with H2O at 110 ℃ under a pressure of 15 MPa, with microwave power of 780 W (PMAE). After centrifugation, the supernatant was collected as the extract. After purification by treatment with absolute alcohol, the analyte in methyl alcohol was used for analysis. Aliquots were taken for qualitative analysis by LC-MS, and for quantitative analysis by HPLC separately. Five alkaloids (i.e., oxymatrine, N-oxysophocarpine, sophoridine, matrine and sophocarpine) were identified by LC-MS. The method of PMAE, as compared with 3 other extraction methods, was proved to be more efficient by having high rate of extraction and requiring less time for extraction. In HPLC analysis, the 5 alkaloids were effectively separated by gradient elution with mobile phase of mixtures of methanol, acetonitrile and 0.1% ammonia solution (pH 10.3) of different ratios on C18 column. Linear relationships between peak area and concentration of the 5 alkaloids were used as basis for quantification. Results of recovery found were in the range of 93.9%-101.0% with RSD′s (n=5) in the range of 0.53%-2.6%.
中图分类号 O652.63
所属栏目 工作简报
基金项目 广州市科技支撑计划项目(No.2008Z1-E301);广东药学院师资队伍建设基金项目(No.52104109)
收稿日期 2011/2/12
修改稿日期
网络出版日期
作者单位点击查看
备注刘小琴(1985-),女,四川泸州人,硕士研究生,主要研究方向为色谱分析。
引用该论文: LIU Xiao-qin,FAN Hua-jun,HUANG Xiao-wen,ZHANG Lai-feng. HPLC Determination of 5 Alkaloids in Sophora Flavescens Ait with Microwave Assisted Extraction[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2012, 48(3): 299~302
刘小琴,范华均,黄晓文,张来凤. 微波辅助提取-高效液相色谱法测定苦参中5种苦参生物碱的含量[J]. 理化检验-化学分册, 2012, 48(3): 299~302
被引情况:
【1】樊轻亚,游国叶, "聚焦微波辅助提取-酸性染料分光光度法测定农吉利中总生物碱",理化检验-化学分册 51, 289-292(2015)
共有人对该论文发表了看法,其中:
人认为该论文很差
人认为该论文较差
人认为该论文一般
人认为该论文较好
人认为该论文很好
参考文献
【1】国家药典委员会.中华人民共和国药典(一部)[M].北京:中国医药科技出版社, 2010:188-189.
【2】李丹,左海军,高慧媛,等.苦参的化学成分[J].沈阳药科大学报, 2004,21(5):346-34.
【3】张少少,安银岭,华燕,等.苦参化学成分和药理研究现状[J].中国民族民间医药, 2008,8(2):8-9.
【4】LIU Xiao-jin, CAO Mei-ai, LI Wen-hai, et al. Alkaloids from sophora flavescens ait.[J]. Fitoterapia, 2010,81(6):524-527.
【5】刘梅,刘雪英,程建峰.苦参碱的药理研究进展[J].中国中药杂志, 2003,28(9):801-804.
【6】衰静,张宗检.苦参碱的生物活性及其研究进展[J].农药, 2003,42(7):1-4.
【7】苗抗立,张建中,董颖,等.苦参的化学成分及药理的研究进展[J].天然产物研究与开发, 2000,13(2):69-73.
【8】秦学功,元英进.苦豆子种子中生物碱的冷浸提取实验研究[J].中草药, 2001,32(7):604-606.
【9】刘勇,熊阳,黄绳武.不同提取方法对苦参生物总碱的影响[J].河南中医学院学报, 2004,19(6):38-39.
【10】邹姝姝,王贵学.中药苦参中苦参碱类成分超声波提取工艺[J].重庆大学学报:自然科学版, 2007,30(7):130-133.
【11】YAN Ming-ming, LIU Wei, FU Yu-jie, et al. Optimisation of the microwave-assisted extraction process for four main astragalosides in Radix Astragali[J]. Food Chemistry, 2010,119(4):1663-1670.
【12】陈浩浩,罗少建,范华均,等.密闭微波辅助提取HPLC测定苦参中苦参碱和氧化苦参碱[J].现代科学仪器, 2010,4(2):108-111.
【13】黄晓文,范华均,黄庆华,等.气相色谱质谱法测定苦参中生物碱[J].理化检验-化学分册, 2009,45(2):190-191,230.
【14】马仁玲,周红华,于喜水,等.苦参药材提取方法的LC/MS研究[J].中成药, 2003,25(10):831-833.
【2】李丹,左海军,高慧媛,等.苦参的化学成分[J].沈阳药科大学报, 2004,21(5):346-34.
【3】张少少,安银岭,华燕,等.苦参化学成分和药理研究现状[J].中国民族民间医药, 2008,8(2):8-9.
【4】LIU Xiao-jin, CAO Mei-ai, LI Wen-hai, et al. Alkaloids from sophora flavescens ait.[J]. Fitoterapia, 2010,81(6):524-527.
【5】刘梅,刘雪英,程建峰.苦参碱的药理研究进展[J].中国中药杂志, 2003,28(9):801-804.
【6】衰静,张宗检.苦参碱的生物活性及其研究进展[J].农药, 2003,42(7):1-4.
【7】苗抗立,张建中,董颖,等.苦参的化学成分及药理的研究进展[J].天然产物研究与开发, 2000,13(2):69-73.
【8】秦学功,元英进.苦豆子种子中生物碱的冷浸提取实验研究[J].中草药, 2001,32(7):604-606.
【9】刘勇,熊阳,黄绳武.不同提取方法对苦参生物总碱的影响[J].河南中医学院学报, 2004,19(6):38-39.
【10】邹姝姝,王贵学.中药苦参中苦参碱类成分超声波提取工艺[J].重庆大学学报:自然科学版, 2007,30(7):130-133.
【11】YAN Ming-ming, LIU Wei, FU Yu-jie, et al. Optimisation of the microwave-assisted extraction process for four main astragalosides in Radix Astragali[J]. Food Chemistry, 2010,119(4):1663-1670.
【12】陈浩浩,罗少建,范华均,等.密闭微波辅助提取HPLC测定苦参中苦参碱和氧化苦参碱[J].现代科学仪器, 2010,4(2):108-111.
【13】黄晓文,范华均,黄庆华,等.气相色谱质谱法测定苦参中生物碱[J].理化检验-化学分册, 2009,45(2):190-191,230.
【14】马仁玲,周红华,于喜水,等.苦参药材提取方法的LC/MS研究[J].中成药, 2003,25(10):831-833.
相关信息