Determination of Quercetin and Kaempferol in Crotalaria ferruginea by QAMS
-
摘要: 为建立适用于响铃草质量控制的槲皮素和山柰素含量的一测多评测定方法，采用高效液相色谱HPLC法，使用C18色谱柱（4.6 mm×250 mm，5 μm），乙腈：0.4%醋酸（40：60）为流动相，检测波长360 nm，柱温30℃，流速1.0 mL·min-1，检测槲皮素和山柰素含量。以槲皮素为内标物，测定其与山柰素的相对校正因子（RCF），并进行含量计算，实现一测多评；同时采用外标法（ESM）测定响铃草中2种有效成分的含量，将一测多评法（QAMS）与外标法（ESM）进行比较。结果表明，8批响铃草药材中山柰素含量的一测多评法测定结果均与外标法无显著性差异。说明该方法操作简便，快速，重复性好，结果准确可靠，可为进一步完善响铃草的质量标准提供参考。Abstract: To establish a QAMS methodology for simultaneous determination of quercetin and kaempferol contents in Crotalaria ferruginea, HPLC was applied on a C18 column (4.6 mm×250 mm, 5 μm) with acetonitrile:0.4% CH3COOH (40:60) for the mobile phase. The column temperature was kept at 30℃ and flow rate at 1.0 mL·min-1. The eluate was detected at the wavelength of 360 nm. Quercetin was used as the internal standard to establish the relative correction factor (RCF) for the quantitative analysis of multi-components on QAMS. The RCF of kaempferol was calculated. The contents of the two compounds were measured by both the external standard method and QAMS to verify the validity of the proposed QAMS methodology. The resulting RCF showed good durability on both quercetin and kaempferol determinations. There were no significant differences between the data obtained by QAMS and by the external standard method. Consequently, this new approach was considered simple, rapid and applicable for quality control on raw material and processed products involving C. ferruginea.
-
Key words:
- Crotalaria ferruginea /
- quercetin /
- kaempferol /
- QAMS /
- HPLC /
- relative correction factors
-
图 1 响铃草中槲皮素和山柰素高效液相色谱
注：A为混合对照品，B为供试品。1为槲皮素，2为山柰素。
Figure 1. HPLC chromatograms for quercetin and kaempferol in Crotalaria ferruginea
表 1 槲皮素和山柰素的加样回收率试验
Table 1. Recovery on quercetin and kaempferol of QAMS method
待测成分 样品量
/g已知量
/mg加入量
/mg测得量
/mg回收率
/%平均回收率
/%RSD
/%槲皮素 3.0005 0.4304 0.4000 0.8281 99.42 98.47 1.19 3.0003 0.4303 0.4000 0.8228 98.12 3.0004 0.4304 0.4000 0.8188 97.1 3.0003 0.4303 0.4000 0.8251 98.71 3.0002 0.4303 0.4000 0.8197 97.34 3.0003 0.4303 0.4000 0.8307 100.1 山柰素 3.0005 0.1667 0.2000 0.1983 98.31 97.88 1.52 3.0003 0.1666 0.2000 0.1967 97.54 3.0004 0.1667 0.2000 0.2023 100.32 3.0003 0.1666 0.2000 0.1949 96.64 3.0002 0.1665 0.2000 0.1939 96.11 3.0003 0.1666 0.2000 0.1983 98.33 
下载: 导出CSV
表 2 不同仪器和色谱柱测定山柰素的相对保留时间
Table 2. RTR of kaempferol by using different instruments and columns
色谱仪器 色谱柱 相对保留时间的比值 Agilent 1260型 Agilent ZORBAX SB-C18 1.525 Phenomenex Gemini C18 1.475 Thermo ODS C18 1.534 赛默飞UltiMate300型 Agilent ZORBAX SB-C18 1.544 Phenomenex Gemini C18 1.553 Thermo ODS C18 1.561 平均值 1.532 RSD/% 2.01
下载: 导出CSV
表 3 响铃草中槲皮素和山柰素的测定
Table 3. Contents of quercetin and kaempferol in C.ferruginea
[单位/(mg · g-1)] 产地 槲皮素ESM 山柰素 ESM QAMS 相对误差/% 广西 0.1431 0.0282 0.0284 0.709 广东 0.0913 0.0312 0.0314 0.641 贵州遵义 0.0392 0.0491 0.0496 1.018 福建 0.0464 0.0783 0.0786 0.383 贵州安顺 0.2712 0.2501 0.2511 0.400 云南 0.0221 0.0795 0.0774 -2.642 浙江 0.0264 0.0414 0.0421 1.691 四川 0.0347 0.0584 0.0576 -1.370 注：相对误差=(QAMS计算值-ESM实测值)/ ESM实测值× 100%
下载: 导出CSV
-
[1] 胡熙明, 张文康, 朱庆生.中华本草[M].上海:上海科学技术出版社, 1999:424. [2] 贵州植物志编辑委员会.贵州植物志(第7卷)[M].四川:四川民族出版社, 1989:381. [3] 唐倩, 杨宪, 刘应杰, 等.响铃草药材中总黄酮和3种黄酮苷元的测定方法及含量变化研究[J].林产化学与工业, 2010, 30(4):93-98. http://www.cqvip.com/qk/93379X/201004/35019569.html [4] 吴文杰, 邓阳, 谭桂林, 等.一测多评法测定葛根药材中5种异黄酮类成分[J].中草药, 2017, 48(4):777-781. http://www.cnki.com.cn/Article/CJFDTOTAL-ZGYX201403016.htm [5] 李文兵, 卢君蓉, 胡麟, 等.一测多评法测定姜黄中姜黄素、去甲氧基姜黄素和双去甲氧基姜黄素[J].中草药, 2017, 48(3):573-577. http://www.cnki.com.cn/Article/CJFDTotal-FJZY201303028.htm [6] 杜憬生, 吴立群, 马鸿雁, 等.一测多评测法测定鸡血藤中4种黄酮成分含量[J].中药材, 2017, 40(4):881-883. http://cbimg.cnki.net/Editor/2018/0212/zyca/06cd4fac-bbe4-4d94-9b9e-64149b062465.pdf [7] 郭燕娜, 胡华杰.一测多评法测定垂盆草颗粒中槲皮素、山柰素与异鼠李素含量[J].中国生化药物杂志, 2016, 36(4):199-202. [8] 曾雪, 杨元娟, 陈竹, 等.HPCE法测定金银花药材中木犀草苷、槲皮素和金丝桃苷的含量[J].中国药房, 2017, 28(18):2543-2545. doi: 10.6039/j.issn.1001-0408.2017.18.27 [9] 周浓, 杨勤, 杨敏, 等.HPLC法同时测定川楝子中芦丁、异槲皮苷和槲皮素的含量[J].药物分析杂志, 2013, 33(2):225-229. [10] 梁国华, 梁琪.HPLC测定复方肝炎颗粒中槲皮素和山奈素的含量[J].中国实验方剂学杂志, 2013, 19(24):90-92. [11] 季宇彬.中药有效成分药理与应用[M].哈尔滨:黑龙江科学技术出版社, 1995:156-157. -
点击查看大图
计量
- 文章访问数: 939
- HTML全文浏览量: 183
- PDF下载量: 7
- 被引次数: 0
