Fang Congbing, Wan Xiaochun, Tan Huarong, Jiang Changjun
Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, Anhui, China.
Ann Chim. 2006 Jan-Feb;96(1-2):117-24. doi: 10.1002/adic.200690002.
Pueraria lobata is a rich source of isoflavonoids. The detection and identification of isoflavonoid components from root, stem, leaf, callus and cell samples, is very important for the best, safest and most efficacious use of kudzu as a medicinal plant, and for the studies on quantitative analysis in the secondary metabolism of isoflavonoids. In this paper, a simple, rapid and precise high-performance capillary electrophoresis (HPCE) method with diode array detection (DAD) has been developed for separation and determination of isoflavonoids in several kudzu samples. The isoflavonoids could be well separated within 15 min in a 40 cm length capillary at a separation voltage of 15kV in a 30 mmol L(-1) borax buffer (pH9.29), and this proposed method demonstrated excellent reproducibility and accuracy with relative standard deviations of less than 5% for isoflavonoid content (n = 5) of different kudzu samples. The relationship between peak areas and isoflavone concentrations, in a specified working range with linear response, was determined by first-order polynomial regression over the range 0.05-0.5 mg mL(-1) for puerarin and 2.5-50 microg mL(-1) for 3'-methoxypuerarin, daidzin and daidzein, respectively, and quantitative evaluation of those four main isoflavonoid components was determined by ultraviolet absorption at lambda = 192 nm. The differences were also illustrated by comparison of the determination of isoflavonoid components from kudzu root, stem, leaf samples and plant tissue cultures in vitro.
葛根是异黄酮的丰富来源。从根、茎、叶、愈伤组织和细胞样本中检测和鉴定异黄酮成分，对于葛根作为药用植物的最佳、最安全和最有效利用，以及对异黄酮次生代谢的定量分析研究非常重要。本文建立了一种简单、快速、精确的带二极管阵列检测（DAD）的高效毛细管电泳（HPCE）方法，用于分离和测定几种葛根样品中的异黄酮。在30 mmol L(-1)硼砂缓冲液（pH9.29）中，在40 cm长的毛细管中，分离电压为15kV时，异黄酮可在15分钟内得到良好分离，该方法对不同葛根样品异黄酮含量（n = 5）的相对标准偏差小于5%，具有出色的重现性和准确性。葛根素在0.05 - 0.5 mg mL(-1)范围内、3'-甲氧基葛根素、大豆苷和大豆黄酮在2.5 - 50 μg mL(-1)范围内，通过一阶多项式回归确定了峰面积与异黄酮浓度在特定工作范围内的线性响应关系，并通过在λ = 192 nm处的紫外吸收对这四种主要异黄酮成分进行了定量评估。通过比较葛根根、茎、叶样品和体外植物组织培养物中异黄酮成分的测定结果，也说明了这些差异。
