Method Development and Validation of 5,7-Dimethoxyflavone Content of Black Ginger Rhizome Aqueous Extract Using Ultra High Performance Liquid Chromatography

Method Development of 5,7-dimethoxyflavone Content of Black Ginger Extract Using UHPLC

Abstract

    Black ginger rhizomes (Kaempferia parviflora Wall. ex Baker) are used for tonic, erectile dysfunction aid, blood circulation system improvement, and antispasmodic in Thai traditional medicine. The major constituent, 5,7-dimethoxyflavone (5,7-DMF), was reported as antioxidant, anti-inflammation and anti-cancer. Until now, the quantitative determination of black ginger rhizomes aqueous extract has not been specifi ed. The objective of this study was to develop and validate method to analyze the content of 5,7-DMF in the extract using ultra high performance liquid chromatography (UHPLC), detected at 264 nm. The stationary phase was ZORBAX Extend-C18 Rapid Resolution HD (2.1×50 mm, 1.8 μm). The mobile phase was the mixture of 0.2% glacial acetic acid in water and methanol as a gradient system. The sample was prepared by sonication with methanol for 30 minutes. As a result, this method was specific. The linearity was shown between 0.06–0.036 mg/ml with r > 0.999. The %recovery was 98.31–105.12. The precision repeatability within day and reproducibility within laboratory, were acceptable with %RSD < 2 and %RSD < 4 (HORRAT < 2), respectively. The limit of quantitation (LOQ) was equal to 0.04 mg/ml. Therefore, the method can be used for determination of 5,7-DMF content for the quality control of black ginger rhizomes aqueous extracts which are commercially available.