Simultaneous Determination of Four Flavone C-Glycosides in Phyllostachys edulis Leaves by High-Performance Liquid Chromatography with Ultraviolet Spectrometry
1. International Centre for Bamboo and Rattan, Beijing 100102, China 2. School of Chemistry and Chemical Engineering, Henan Institute of Science and Technology, Xinxiang 453003, China 3. School of Resources and Environment, Anhui Agricultural University, Hefei 230036, China
Abstract：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×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 ℃, 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.
[1] HE Yue-jun，YUE Yong-de(何跃君，岳永德). Biomass Chemical Engineering(生物质化学工程)，2008，42(3)：31. [2] GONG Jin-yan，WU Xiao-qin，ZHANG Ying(龚金炎，吴晓琴，张 英). Natural Product Research and Development(天然产物研究与开发)，2010，22: 525. [3] WANG Wen-yuan(王文渊). Food and Nutrition in China(中国食物与营养)，2012，18(6)：71. [4] Zhang Yu，Bao Bili，Lu Boyi，et al. Journal of Chromatography A，2005，1065(2)：177. [5] LI Hong-yu，SUN Jing-yun，ZHANG Jian-min，et al(李洪玉，孙静芸，张建民，等). Chinese Traditional Patent Medicine(中成药)，2004，26(3)：208. [6] GONG Jin-yan，WU Xiao-qin，XIA Dao-zong，et al(龚金炎，吴晓琴，夏道宗，等). Chinese Traditional and Herbal Drugs(中草药)，2010，41(1)：63. [7] Guo Xuefeng，Yue Yongde，Tang Feng，et al. Journal of Food Biochemistry，2013，30(4)，doi：10.1111/j.1745-4514.2012.00674.x. [8] WU Yan-shu，JIN Cheng, ZHANG Qian，et al(武彦舒，金 城，张 倩，等). Spectroscopy and Spectral Analysis(光谱学与光谱分析)，2009，29(11)：3112. [9] CHEN Yan，YAO Hong，LI Shao-guang，et al(陈 艳，姚 宏，李少光，等). Chinese Journal of Pharmaceutical Analysis(药物分析杂志)，2011，31(4)：645. [10] Wang Jin，Yue Yongde，Jiang Hao，et al. International Journal of Analytical Chemistry，2012，doi：10.1155/2012/205101. [11] HE Yu-fang，NAN Min-lun，ZHAO Yu-wei，et al(赫玉芳，南敏伦，赵昱玮，等). Journal of Shenyang Pharmaceutical University(沈阳药科大学学报)，2013，30(3)：197. [12] JI Ri-li，ZHU Zheng-ying，ZHAO Li-hua，et al(季日丽，朱整英，赵丽华，等). China Pharmaceuticals(中国药业)，2012，21(8)：44. [13] WU De-ling，JIN Chuan-shan，LI Qiong，et al(吴德玲，金传山，李 琼，等). Journal of Chinese Medicinal Materials(中药材)，2011，34(10)：1556.
