Development and Validation of an HPLC Method for Quality Control of Pueraria lobata Flower

Abstract

Pueraria lobata, also known as Kudzu (Japan) or Ge (China), is a medicinal plant widely used in Oriental traditional medicine. In this study the development, optimization and validation of an HPLC ethod for quality control of Pueraria flower plant material is presented. By means of this analytical method the three major compounds, i. e., the isoflavones tectorigenin 7-O-[β-D-xylopyranosyl-(1 - 6)-β-D-glucopyranoside], tectorigenin 7-O-β-D-glucopyranoside and tectorigenin, were quantified, using the isoflavones genistin and genistein as external standards. The extraction procedure, the extraction solvent, the extraction yields and the HPLC conditions were evaluated and optimized. The samples were analyzed on an RP C18 column, and eluted with a binary system consisting of water and methanol using a linear gradient; detection was at 262 nm. Tectorigenin used in the recovery experiments was isolated and purified in the laboratory. The final method was fully validated according to the ICH guidelines in terms of linearity, precision and accuracy. The validation data showed that the precision, (RSD%_{between days} of 3.1, 2.84 and 1.77 for the three major compounds, respectively), and the accuracy (recovery of 104.2 %) were acceptable. These validation results demonstrate the suitability of the method for the quality control of this crude drug.

Abbreviations

ANOVA:analysis of variance

Cnf. limit:confidence limit

DAD:diode array detector

F crit:F critical

F LOF:F lack of fit

ICH:International Conference on Harmonization

RSD:relative standard deviation

Key words

Pueraria lobata - Fabaceae - flowers - HPLC analysis - isoflavonoids

References

• 1 Keung W M. The genus Pueraria (Ge) in traditional Chinese herbal medicine, 1^st edition. Oxfordshire: Taylor & Francis 2002: 59-69
• 2 Miyazawa M, Sakano K, Nakamura S, Kosaka H. Antimutagenic activity of isoflavones from Pueraria lobata .  J Agric Food Chem. 2001;  49 336-41.
• 3 Kinjo J, Takeshita T, Abe Y, Terada N, Yamashita H, Yamasaki M. et al . Studies on the constituents of Pueraria lobata. Chemical constituents in the flowers and the leaves.  Chem Pharm Bull. 1988;  36 1174-9.
• 4 European Directorate for the Quality of Medicines. European Pharmacopoeia, 5th edition. Strasbourg; EDQM 2005: 1433.
• 5 Aida M, Hano Y, Nomura T. Ficusins A and B, two new cyclic-monoterpene-substituted isoflavones from Ficus septica Barm. F.  Heterocycles. 1995;  41 2761-8.
• 6 Park H J, Park J H, Moon J, Lee K T, Jung W T, Oh S R. et al . Effects of naturally occurring isoflavones on prostaglandin E2 production.  Phytochemistry. 1999;  51 147-51.
• 7 Ding H, Chen Y, Chang W, Lin H. Flavonoids from the flowers of Pueraria lobata .  J Chin Chem Soc. 2004;  51 1425-8.
• 8 Text on Validation of Analytical Procedures - ICH Harmonized Tripartite Guideline. ICH 1994.
• 9 Validation of Analytical Procedures: Methodology - ICH Harmonized Tripartite Guideline. ICH 1996.
• 10 Caporal-Gautier J, Nivet J M, Algranti P, Guiloteau M, Histe M, Lallier M. et al . Guide de validation analytique. Rapport d’une commission SFSTP. I. Methodologie.  Stp Pharma Prat. 1992;  4 201-26.