Su Juan, Fu Peng, Shen Yunheng, Zhang Chuan, Liang Mingjin, Liu Runhui, Li Huiliang, Zhang Weidong
Department of Natural Medicinal Chemistry, School of Pharmacy, Second Military Medical University, Shanghai 200433, PR China.
J Pharm Biomed Anal. 2008 Jan 22;46(2):342-8. doi: 10.1016/j.jpba.2007.10.032. Epub 2007 Nov 6.
A novel and sensitive HPLC-UV method has been developed for the simultaneous determination of five major flavonoids in Hypericum japonicum hydroalcoholic extract. The chemical profile of five flavonoids, including taxfolin-7-O-alpha-L-rhamnoside (1), isoquercitrin (2), quercitrin (3), quercetin-7-O-alpha-L-rhamnoside (4) and quercetin (5) was acquired by using high-performance liquid chromatography-diode array detector coupled to an electrospray tandem mass spectrometer (HPLC-DAD-ESI/MS). The analysis was performed on a ZORBAX SB-C18 analytical column (5 microm, 250 mm x 4.6 mm, i.d.) with a gradient solvent system of acetonitrile-0.5% aqueous formic acid. The validation was carried out and the linearities (r(2)>0.9997) and recoveries (ranged from 98.4% to 99.8%) were acceptable. The limits of detection (LOD) of these flavonoids ranged from 0.5 to 7.5 ng. The results indicated that the contents of investigated flavonoids in H. japonicum varied significantly from habitat to habitat with contents ranging from 2.00 to 34.18 mg/g. The proposed method is simple, effective and suitable for the quality control of this traditional Chinese medicine (TCM).
已开发出一种新颖且灵敏的高效液相色谱 - 紫外检测法，用于同时测定地耳草水醇提取物中的五种主要黄酮类化合物。使用高效液相色谱 - 二极管阵列检测器与电喷雾串联质谱仪（HPLC - DAD - ESI/MS）联用，获得了五种黄酮类化合物的化学图谱，包括紫杉叶素 - 7 - O - α - L - 鼠李糖苷（1）、异槲皮苷（2）、槲皮苷（3）、槲皮素 - 7 - O - α - L - 鼠李糖苷（4）和槲皮素（5）。分析在ZORBAX SB - C18分析柱（5μm，250mm×4.6mm，内径）上进行，采用乙腈 - 0.5%甲酸水溶液的梯度溶剂系统。进行了方法验证，线性关系（r²>0.9997）和回收率（范围为98.4%至99.8%）均可接受。这些黄酮类化合物的检测限（LOD）范围为0.5至7.5 ng。结果表明，地耳草中所研究的黄酮类化合物含量因产地不同而有显著差异，含量范围为2.00至34.18 mg/g。所提出的方法简单、有效，适用于这种中药的质量控制。
