Zhang Chun-Juan, Meng Zhi-Fen, Guo Xue-Feng, Yue Yong-De, Tang Feng, Yu Jin, Chen Chao
Guang Pu Xue Yu Guang Pu Fen Xi. 2014 Sep;34(9):2568-72.
High-performance liquid chromatography with ultraviolet spectrometer (HPLC-UV) was used to simultaneously detect the four flavone C-glycosides, i. e. orientin, isoorientin, vitexin and isovitexin. Analytes were separated with Waters XTerra MS C18 column (250 mm x 4.6 mm, 5 μm) using acetonitrile and 0.5% (φ) formic acid as mobile phase. The flow rate was set at 1.0 mL · min(-1) with the column temperature at 30 °C, and the detection wavelength was 360 nm. The calibration curve was linear over the concentration range of 0.1-10.0 mg · L(-1) for the mixed standard solution. Analytes were separated in 22 minutes, and the relative standard deviation values were all above 0.999. LOD values of standards were found to be between 0.03 and 0.07 mg · L(-1), and LOQ values were in the range of 0.04-0.08 mg · L(-1). After comparing the spectra (240-400 nm) of four flavone C-glycosides in mixed standards and the final product purified by macroporous resin, respectively, the curve shape and characteristic ultraviolet absorption wavelength of each flavone C-glycoside including orientin, isoorientin, vitexin and isovitexin were fitted well. The bamboo leaves sample was extracted by ethanol under reflux, and then partitioned with water and petroleum ether. The aqueous phase was added onto macroporous resin (AB-8), and the fraction of ethanol-water (40%, φ) was concentrated. It was found that the contents of orientin, isoorientin, vitexin and isovitexin relative to the fraction of ethanol- water were 13.73, 49.68, 7.85 and 30.70 mg · g(-1), respectively. In addition, the average recovery of the four flavone C-glycosides ranged from 34.90% to 87.64% with RSD values from 0.41% to 10.83%. The results showed that bamboo leaves sample had good stability and repeatability. The new method was used to analyze the four flavone C-glycosides quickly and provide quality control for commercial products.
采用高效液相色谱-紫外光谱联用仪(HPLC-UV)同时检测4种黄酮碳苷,即荭草苷、异荭草苷、牡荆苷和异牡荆苷。采用Waters XTerra MS C18色谱柱(250 mm×4.6 mm,5 μm),以乙腈和0.5%(φ)甲酸为流动相分离分析物。流速设定为1.0 mL·min-1,柱温为30℃,检测波长为360 nm。混合标准溶液在0.1~10.0 mg·L-1浓度范围内校准曲线呈线性。分析物在22分钟内分离,相对标准偏差值均高于0.999。标准品的检测限在0.03~0.07 mg·L-1之间,定量限在0.04~0.08 mg·L-1范围内。分别比较混合标准品和大孔树脂纯化后最终产物中4种黄酮碳苷的光谱(240~400 nm),荭草苷、异荭草苷、牡荆苷和异牡荆苷等各黄酮碳苷的曲线形状和特征紫外吸收波长拟合良好。竹叶样品用乙醇回流提取,然后用水和石油醚萃取。水相上样到大孔树脂(AB-8)上,浓缩乙醇-水(40%,φ)馏分。发现荭草苷、异荭草苷、牡荆苷和异牡荆苷相对于乙醇-水馏分的含量分别为13.73、49.68、7.85和30.70 mg·g-1。此外,4种黄酮碳苷的平均回收率在34.90%~87.64%之间,相对标准偏差值在0.41%~10.83%之间。结果表明竹叶样品具有良好的稳定性和重复性。该新方法可快速分析4种黄酮碳苷,为商品提供质量控制。