title: Separation and determination of isoflavonoids in several kudzu samples by high-performance capillary electrophoresis (HPCE).
abstract: 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.
authors.affiliation: Key Laboratory of Tea Biochemistry and Biotechnology, Ministry of Education and Ministry of Agriculture, Anhui Agricultural University, Hefei 230036, Anhui, China.
authors.forename: Congbing
authors.initials: C
authors.lastname: Fang
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Wan
authors.forename: Huarong
authors.initials: H
authors.lastname: Tan
authors.forename: Changjun
authors.initials: C
authors.lastname: Jiang
mesh_terms: D001881:Borates / Q000737:chemistry; D002021:Buffers; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D019075:Electrophoresis, Capillary / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007529:Isoflavones / Q000737:chemistry / Q000302:isolation & purification*; D018515:Plant Leaves / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D018547:Plant Stems / Q000737:chemistry; D018514:Plant Structures / Q000737:chemistry; D010946:Plants, Medicinal / Q000737:chemistry*; D029908:Pueraria / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D046509:Tissue Culture Techniques
chemical_list: D001881:Borates; D002021:Buffers; D007529:Isoflavones; C018851:borax; C013908:daidzin; C004742:daidzein