RP-HPLC法同时测定野葛的根、茎和叶中葛根素、大豆苷和大豆苷元的含量

doi:10.7506/spkx1002-6630-200914054

食品科学 ›› 2009, Vol. 30 ›› Issue (14): 248-252.doi: 10.7506/spkx1002-6630-200914054

RP-HPLC法同时测定野葛的根、茎和叶中葛根素、大豆苷和大豆苷元的含量

吴向阳¹，仰玲玲²，仰榴青^{3 ,*} ，邹艳敏³，陆继明⁴

1.江苏大学化学化工学院 2. 江苏大学食品与生物工程学院 3.江苏大学药学院 4. 镇江缘源葛业开发有限公司

收稿日期:2008-10-10 修回日期:2009-01-11 出版日期:2009-07-15 发布日期:2010-12-29
通讯作者: 仰榴青 E-mail:foodwhat605b@yahoo.com.cn

Simultaneous RP-HPLC Determination of Puerarin, Daidzin and Daidzein in Roots, Stems and Leaves of Pueraria lobata (Wild) Ohwi

WU Xiang-yang1，YANG Ling-ling2，YANG Liu-qing3,*，ZOU Yan-min3，LU Ji-ming4

1. College of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang 212013, China；2. College of Food and Biological
Engineering, Jiangsu University, Zhenjiang 212013, China； 3. College of Pharmacy, Jiangsu University, Zhenjiang 212013,
China；4. Zhenjiang Yuanyuan Geye Development Co. Ltd., Zhenjiang 212003, Chi

Received:2008-10-10 Revised:2009-01-11 Online:2009-07-15 Published:2010-12-29
Contact: YANG Liu-qing3,*， E-mail:foodwhat605b@yahoo.com.cn

摘要/Abstract

摘要：

目的：建立RP-HPLC 法同时测定野葛的根、茎及叶中葛根素、大豆苷和大豆苷元的方法。方法：样品前处理采用70% 乙醇超声提取，HPLC 直接测定，并与药典方法进行对比。结果：对照品葛根素、大豆苷和大豆苷元分别在11.87～73.91，1.00～6.48 和0.28～1.53μg/ml 范围内呈良好线性关系，相关系数 r 分别为0.9998，0.9997 和0.9994，最低检出限分别为0.43、0.24、0.18μg/ml。平均回收率分别为99.78%、102.61% 和105.57%，RSD 分别为1.6%、2.8% 和3.4%(n = 5)。精密度、稳定性和重现性良好。超声提取法测得的葛根中葛根素、大豆苷和大豆苷元含量分别为3.98%、0.36% 和0.039%，高于药典提取法的提取率，更适于同时提取葛根素、大豆苷和大豆苷元。葛茎中葛根素、大豆苷和大豆苷元含量分别为0.004%、0.0032% 和0.0007%，葛叶中葛根素、大豆苷和大豆苷元含量分别为0.0016%、0.005% 和0.0012%。结论：该方法准确、快速，操作简单，分离效果好，可用于野葛的根、茎和叶中葛根素、大豆苷和大豆苷元含量的同时测定；江苏茅山地区野葛茎和叶中葛根素、大豆苷和大豆苷元含量大大低于葛根中的含量。

关键词: 野葛, 葛根素, 大豆苷, 大豆苷元, 高效液相色谱法

Abstract:

A RP-HPLC method was established for the simultaneous determination of puerarin, daidzin and daidzein in roots, stems and leaves of Pueraria lobata (Wild) Ohwi. The procedure was based on extraction of test portions with 70% ethanol solution in an ultrasonic field for 20 min at 50 ℃ followed by direct HPLC analysis of the extracts. The chromatographic conditions were as follows: using a Hypersil ODS-2 column (4.6 mm × 250 mm, 5 μm) and a mobile phase composed of methanol and H2O, gradient elution [0－4 min 25:75 (methanol/water volume ratio), 4－7 min 25:75 → 32:78, 7－9min 32:68, 9－11 min 32:68 → 50:50, 11－24 min 50:50, 24－30 min 50:50 → 25:75], column temperature 35 ℃, detection wavelength 250 nm, and flow rate of the mobile phase 0.8 ml/min. A good linear relationship between peak area and concentration was found to puerarin, daidzin and daidzein in the ranges of 11.87－73.91 (r = 0.9998), 1.00－6.48 (r = 0.9997) and 0.28－1.53 μg/ml (r = 0.9994), respectively. The LODs were 0.43, 0.24 and 0.18μg/ml, respectively. Mean recoveries were 99.78%, 102.61% and 105.57% with the RSDs of 1.6%, 2.8% and 3.4% (n = 5), respectively. The precision, stability and reproducibility of the method all were good. The contents of puerain, daidzin, and daidzein in roots of Pueraria lobata (wild) Ohwi determined by this method were 3.98%, 0.36% and 0.039% respectively, higher than 3.28%, 0.26% and 0.028% of those analyzed according to the method stipulated in the Chinese Pharmacopoeia 2005 respectively, suggesting high efficiency of ultrasonic-assisted extraction of the three analytes. The contents of puerain, daidzin, and daidzein in stems of Pueraria lobata (wild) Ohwi were 0.004%, 0.0032% and 0.0007% respectively, while those in leaves were 0.0016%, 0.005% and 0.0012%, respectively. This method is accurate, fast and simple and therefore can be used for the simultaneous determination of puerain, daidzin, and daidzein in roots, stems and leaves of Pueraria lobata (wild) Ohwi. Our results demonstrate that the contents of puerain, daidzin, and daidzein in stems and leaves of Pueraria lobata (wild) Ohwi growned in Maoshan area of Jiangsu province are obviously lower than those in the roots.

Key words: Pueraria lobata (wild) Ohwi, puerarin, daidzin, daidzein, HPLC

中图分类号:

R284.1

吴向阳1，仰玲玲2，仰榴青3,*,邹艳敏3，陆继明4. RP-HPLC法同时测定野葛的根、茎和叶中葛根素、大豆苷和大豆苷元的含量[J]. 食品科学, 2009, 30(14): 248-252.

WU Xiang-yang1，YANG Ling-ling2，YANG Liu-qing3,*，ZOU Yan-min3，LU Ji-ming4. Simultaneous RP-HPLC Determination of Puerarin, Daidzin and Daidzein in Roots, Stems and Leaves of Pueraria lobata (Wild) Ohwi[J]. FOOD SCIENCE, 2009, 30(14): 248-252.

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RP-HPLC法同时测定野葛的根、茎和叶中葛根素、大豆苷和大豆苷元的含量

Simultaneous RP-HPLC Determination of Puerarin, Daidzin and Daidzein in Roots, Stems and Leaves of Pueraria lobata (Wild) Ohwi

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