一测多评HPLC法测定三叶青中5种黄酮类成分的含量
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摘要
目的:建立HPLC一测多评法(QAMS)同时测定三叶青中芦丁、异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素、山奈酚5种黄酮类成分。方法:色谱柱:Agilent ZORBAX SB-C_(18)(250 mm×4. 6 mm,5μm);流动相:乙腈-0. 2%磷酸溶液,梯度洗脱;流速:1. 0 ml·min~(-1);检测波长:360 nm;柱温:35℃;进样量:10μl。以芦丁为内参物,分别建立其与异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素、山奈酚间的相对校正因子(RCF),并计算三叶青各成分含量,实现一测多评。结果:芦丁与异槲皮苷、山奈酚-3-O-芸香糖苷、槲皮素和山奈酚的RCF分别为0. 918,3. 922,0. 082,0. 088,RSD分别为1. 30%,0. 63%,3. 88%,1. 12%(n=13);17批三叶青中以芦丁为参照物,另外4种黄酮类成分的含量用一测多评法测定结果与外标法测定结果相对误差<5%。结论:以芦丁为参照,采用QAMS能同时测定三叶青中5种黄酮类成分的含量,该方法可靠,结果准确,可用于三叶青药材的质量控制。
Objective: To determine 5 major compounds (rutin, isoquercitrin, kaempferol-3-O-rutinoside, quercetin and kaempferol) by quantitative analysis of multi-components with single maker,and the feasibility and technical adaptability of the method were verified in Tetrastigma hemsleyanum. Methods: The column was Agilent ZORBAX SB-C_(18) (250 mm × 4. 6 mm,5μm),the mobile phase was acetmitrilc-0. 2% phosphcric acid solution (gradient edution),the delection wavelength was 360 nm,the temperature was 35℃,the flow velocity was 1. 0 ml·min~(-1),the sample size was 10 μl. The relative correction factors of isoquercitrin,kaempferol-3-Orutinoside,quercetin and kaempferol were determined with rutin as the internal standard. And the content of each component was calculated in Tetrastigma hemsleyanum. At the same time,the external standard method was used to verify the accuracy and feasibility of the multi-components with single maker. Results: The relative correction factors of rutin,isoquercitrin,kaempferol-3-O-rutinoside,quercetin and kaempferol was 0. 918,3. 922,0. 082 and 0. 088 with the RSD of 1. 30,0. 63,3. 88 and 1. 12 (n = 13),respectirel. Rutin was used as the reference substance in 17 batches of Tetrastigma hemsleyanum. The relative error of the contents of the four flavonoids in the 17 batches of Tetrastigma hemsleyanum was less than 5%. Conclusion: The method is simple,quick and accurate,and suitable for the quality control of Tetrastigma hemsleyanum.
Objective: To determine 5 major compounds (rutin, isoquercitrin, kaempferol-3-O-rutinoside, quercetin and kaempferol) by quantitative analysis of multi-components with single maker,and the feasibility and technical adaptability of the method were verified in Tetrastigma hemsleyanum. Methods: The column was Agilent ZORBAX SB-C_(18) (250 mm × 4. 6 mm,5μm),the mobile phase was acetmitrilc-0. 2% phosphcric acid solution (gradient edution),the delection wavelength was 360 nm,the temperature was 35℃,the flow velocity was 1. 0 ml·min~(-1),the sample size was 10 μl. The relative correction factors of isoquercitrin,kaempferol-3-Orutinoside,quercetin and kaempferol were determined with rutin as the internal standard. And the content of each component was calculated in Tetrastigma hemsleyanum. At the same time,the external standard method was used to verify the accuracy and feasibility of the multi-components with single maker. Results: The relative correction factors of rutin,isoquercitrin,kaempferol-3-O-rutinoside,quercetin and kaempferol was 0. 918,3. 922,0. 082 and 0. 088 with the RSD of 1. 30,0. 63,3. 88 and 1. 12 (n = 13),respectirel. Rutin was used as the reference substance in 17 batches of Tetrastigma hemsleyanum. The relative error of the contents of the four flavonoids in the 17 batches of Tetrastigma hemsleyanum was less than 5%. Conclusion: The method is simple,quick and accurate,and suitable for the quality control of Tetrastigma hemsleyanum.
引文
1全国中草药汇编编写组.全国中草药汇编.上册[M].北京:人民卫生出版社,1975:31
2许文,傅志勤,林婧,等.UPLC-MS/MS法同时测定三叶青中10种黄酮类成分[J].药学学报,2014,49(12):1711-1717
3许文,傅志勤,林婧,等.HPLC-Q-TOF-MS和UPLC-Qq Q-MS的三叶青主要成分定性与定量研究[J].中国中药杂志,2014,39(22):4365
4范世明,徐惠龙,谢心月,等.三叶青叶指纹图谱研究及8种酚类成分含量测定[J].中国中药杂志,2016,21(44):3975-3981
5浦锦宝,梁卫青,郑军献,等.HPLC法测定三叶青中槲皮素、山奈酚的含量[J].中国中医药科技,2011,18(2):134-135
6刘宇文,殷红妹,邹耀华.HPLC-ELSD法测定三叶青中的β-谷甾醇的含量[J].中华中医药学刊,2012,30(6):1387-1389
7丁丽,章璐幸,邱彦.HPLC-ELSD法同时测定三叶青中胡萝卜苷和β-谷甾醇的含量[J].安徽医药,2015,19(11):2083-2084
8王智民,高慧敏,付雪涛,等.“一测多评法”中药质量评价模式方法学研究[J].中国中药杂志,2006,31(23):1925-1928
9王智民,钱忠直,张启伟,等.一测多评法建立的技术指南[J].中国中药杂志,2011,36(6):657-658
10黄容妹,刘伟锐,刘翠红,等.一测多评法测定银杏叶中总内酯含量[J].今日药学,2016,26(4):231-234
11徐容.一测多评法同时测定妇乐颗粒中4种成分的含量[J],中国现代应用药学,2017,34(3):403-406
2许文,傅志勤,林婧,等.UPLC-MS/MS法同时测定三叶青中10种黄酮类成分[J].药学学报,2014,49(12):1711-1717
3许文,傅志勤,林婧,等.HPLC-Q-TOF-MS和UPLC-Qq Q-MS的三叶青主要成分定性与定量研究[J].中国中药杂志,2014,39(22):4365
4范世明,徐惠龙,谢心月,等.三叶青叶指纹图谱研究及8种酚类成分含量测定[J].中国中药杂志,2016,21(44):3975-3981
5浦锦宝,梁卫青,郑军献,等.HPLC法测定三叶青中槲皮素、山奈酚的含量[J].中国中医药科技,2011,18(2):134-135
6刘宇文,殷红妹,邹耀华.HPLC-ELSD法测定三叶青中的β-谷甾醇的含量[J].中华中医药学刊,2012,30(6):1387-1389
7丁丽,章璐幸,邱彦.HPLC-ELSD法同时测定三叶青中胡萝卜苷和β-谷甾醇的含量[J].安徽医药,2015,19(11):2083-2084
8王智民,高慧敏,付雪涛,等.“一测多评法”中药质量评价模式方法学研究[J].中国中药杂志,2006,31(23):1925-1928
9王智民,钱忠直,张启伟,等.一测多评法建立的技术指南[J].中国中药杂志,2011,36(6):657-658
10黄容妹,刘伟锐,刘翠红,等.一测多评法测定银杏叶中总内酯含量[J].今日药学,2016,26(4):231-234
11徐容.一测多评法同时测定妇乐颗粒中4种成分的含量[J],中国现代应用药学,2017,34(3):403-406