Li Chenrui, Zhou Limin, Lin Ge, Zuo Zhong
School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
J Pharm Biomed Anal. 2009 Oct 15;50(3):298-306. doi: 10.1016/j.jpba.2009.04.028. Epub 2009 May 5.
A simple and efficient HPLC/UV method for the simultaneous determination of six bioactive flavones, namely baicalein, baicalin, wogonin, wogonoside, oroxylin A and oroxylin A-7-O-glucuronide, has been developed and applied for their content determination in reference herb and proprietary traditional Chinese medicine (PTCM) products of radix Scutellariae. The chromatographic separation was carried out on a Thermo C(18) column and linear gradient elution was employed with a mobile phase containing acetonitrile and 20 mM sodium dihydrogen phosphate buffer (pH 4.6). All the analytes were detected by PDA detector at a wavelength of 270 nm. Contents of the analytes in radix Scutellariae containing PTCM products in forms of capsule, soft capsule, tablet and dripping pill and the reference herb of radix Scutellariae were analyzed by sonicator extraction with methanol and water mixture (80:20) containing 1 mM HCl for 30 min followed by HPLC analysis. Separation of the six analytes was achieved within 25 min with good linearity (R(2)>0.99). The R.S.D. of both the intra-day and inter-day precision for all the six analytes was below 10.14%. The accuracy at different concentrations was within the range of -7.83 to 4.06%. The extraction recovery was within the range of 89.22-107.33% for all the analytes. Contents of the six flavones were found to vary significantly among different products with glycosides, such as baicalin, wogonoside and oroxylin A-7-O-glucuronide, in much greater quantity than their corresponding aglycones. In addition to baicalin (18.54+/-0.71%, w/w), the commonly used marker compound for radix Scutellariae, wogonoside (3.54+/-0.18%, w/w) and oroxylin A-7-O-glucuronide (2.84+/-0.14%, w/w) also existed in abundant amount in the reference herb. Our findings suggested that wogonoside and oroxylin A-7-O-glucuronide should also be served as the chemical markers together with baicalin for the quality control of herbs and PTCM products of radix Scutellariae.
已开发出一种简单高效的HPLC/UV方法,用于同时测定六种生物活性黄酮,即黄芩素、黄芩苷、汉黄芩素、汉黄芩苷、木犀草素和木犀草素-7-O-葡萄糖醛酸苷,并将其应用于黄芩对照药材和中成药产品中这些成分的含量测定。色谱分离在Thermo C(18)柱上进行,采用线性梯度洗脱,流动相为含乙腈和20 mM磷酸二氢钠缓冲液(pH 4.6)的混合液。所有分析物均通过PDA检测器在270 nm波长处进行检测。采用含1 mM HCl的甲醇和水混合液(80:20)超声提取30 min后进行HPLC分析,测定了胶囊、软胶囊、片剂和滴丸形式的含黄芩中成药产品以及黄芩对照药材中分析物的含量。六种分析物在25 min内实现分离,线性关系良好(R(2)>0.99)。六种分析物日内和日间精密度的相对标准偏差均低于10.14%。不同浓度下的准确度在-7.83至4.06%范围内。所有分析物的提取回收率在89.22 - 107.33%范围内。发现六种黄酮的含量在不同产品中差异显著,糖苷形式如黄芩苷、汉黄芩苷和木犀草素-7-O-葡萄糖醛酸苷的含量远高于其相应的苷元。除了常用的黄芩苷(18.54±0.71%,w/w)作为黄芩的标记化合物外,汉黄芩苷(3.54±0.18%,w/w)和木犀草素-7-O-葡萄糖醛酸苷(2.84±0.14%,w/w)在对照药材中也大量存在。我们的研究结果表明,汉黄芩苷和木犀草素-7-O-葡萄糖醛酸苷也应与黄芩苷一起作为黄芩药材和中成药产品质量控制的化学标记物。