食品科学

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湘葛一号茎叶中5 种异黄酮成分的高效液相色谱法定量分析

叶惠煊,李 芝,李小军,黄玮超,谢 霞,刘向前*   

  1. 湖南中医药大学药学院,湖南省中药现代化研究重点实验室,湖南 长沙 410208
  • 出版日期:2014-07-25 发布日期:2014-08-26
  • 通讯作者: 刘向前
  • 基金资助:

    湖南省自然科学基金重点项目(11JJ2042);长沙市科技局科技计划重点项目(K1207010-21);
    湖南中医药大学药物分析学“十二五”校级重点学科建设项目(2012-2)

Simultaneous Determination of Five Isoflavones in Stems and Leaves of Pueraria thomsonii Benth ‘Xiangge No.1’ by HPLC

YE Hui-xuan, LI Zhi, LI Xiao-jun, HUANG Wei-chao, XIE Xia, LIU Xiang-qian*   

  1. Modernization Laboratory of Traditional Chinese Medicine and Drug, College of Pharmacy,
    Hunan University of Traditional Chinese Medicine, Changsha 410208, China
  • Online:2014-07-25 Published:2014-08-26
  • Contact: LIU Xiang-qian

摘要:

建立高效液相色谱法同时测定湘葛一号地上部分中5 种异黄酮成分含量的方法。采用AT·LICHROM C18
色谱柱(4.6 mm×250 mm,5 μm),以甲醇-水为流动相进行梯度洗脱,流速1.0 mL/min,检测波长250 nm,
柱温30 ℃。葛根素、大豆苷、染料木苷、大豆苷元和染料木素分别在5.00~50.00(r=0.999 4)、1.00~10.00
(r=0.999 2)、3.64~36.40(r=0.999 9)、0.50~5.00 μg/mL(r=0.999 8)和0.10~1.00 μg/mL(r=0.999 4)范围
内线性良好,检测限分别为0.94、0.24、0.43、0.08 ng/L和0.05 ng/L,定量限分别为2.85、0.73、1.29、0.25 ng/L
和0.15 ng/L。精密度、稳定性和重复性良好,平均回收率分别为100.11%(相对标准偏差(RSD)为1.39%)、
99.54%(RSD为1.47%)、98.06%(RSD为0.32%)、101.52%(RSD为1.89%)和101.05%(RSD为0.32%)。该方
法快速、准确,操作简单,可用于湘葛一号地上部分5 种异黄酮成分含量的同时测定。

关键词: 湘葛一号, 高效液相色谱法, 葛根素, 大豆苷, 染料木苷, 大豆苷元, 染料木素

Abstract:

An HPLC method was presented for the simultaneous determination of five isoflavones simultaneously in the
aerial (leaves and stems) parts of Pueraria thomsonii Benth ‘Xiangege No.1’. The chromatography was performed on an
AT·LICHROM C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase of methanol-water by gradient elution at a flow
rate of 1.0 mL/min, and the detection wavelength was set at 250 nm and the column temperature at 30 ℃. The calibration
curves for puerarin, daidzin, genistin, daidzein and genistein were linear in the range of 5.00–50.00 (r = 0.999 4), 1.00–10.00
(r = 0.999 2), 3.64–36.40 (r = 0.999 9), 0.50–5.00 (r = 0.999 8) and 0.10–1.00 μg/mL (r = 0.999 4), respectively. The
LODs were 0.94, 0.24, 0.43, 0.08 and 0.05 ng/L, and the LOQs were 2.85, 0.73, 1.29, 0.25 and 0.15 ng/L. The method had
good precision, stability and reproducibility, and the mean recoveries for puerarin, daidzin, genistin, daidzein and genistein
were 100.11% (RSD = 1.39%), 99.54% (RSD = 1.47%), 98.06% (RSD = 0.32%), 101.52% (RSD = 1.89%) and 101.05%
(RSD = 0.32%), respectively. In conclusion, this study has proposed a rapid, accurate and simple method which allows the
simultaneous determination of five isoflavones in the aerial parts of Pueraria thomsonii Benth.

Key words: Hu’nan Pueraria thomsonii Benth, high performance liquid chromatography, puerarin, daidzin, genistin, daidzein, genistein

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