A Simple Isocratic HPLC Method for the Simultaneous Determination of Sinensetin, Eupatorin, and 30-hydroxy-5,6,7,4'-tetramethoxyflavone in Orthosiphon stamineus Extracts
- Authors
- Mun Fei Yam; Elsnoussi Ali Hussin Mohamed; Lee Fung Ang; Li Pei; Yusrida Darwis; Roziahanim Mahmud; Mohd. Zaini Asmawi; Rusliza Basir; Mariam Ahmad; Yoo, Hye Hyun; KIM DONG HYUN; Chung, Hye Jin
- Issue Date
- Jun-2012
- Publisher
- 사단법인약침학회
- Keywords
- 3' -hydroxy-5; 6; 7; 4'- tetramethoxyflavone; eupatorin; flavonoids; O stamineus; HPLC; sinensetin
- Citation
- Journal of acupuncture and meridian studies, v.5, no.4, pp 176 - 182
- Pages
- 7
- Indexed
- SCOPUS
KCICANDI
- Journal Title
- Journal of acupuncture and meridian studies
- Volume
- 5
- Number
- 4
- Start Page
- 176
- End Page
- 182
- URI
- https://scholarworks.bwise.kr/erica/handle/2021.sw.erica/35045
- DOI
- 10.1016/j.jams.2012.05.005
- ISSN
- 2005-2901
2093-8152
- Abstract
- Orthosiphon stamineus extracts contain three flavonoids (3’-hydroxy-5, 6, 7, 4’-tetramethoxyflavone, sinensetin, and eupatorin) as bioactive substances. Previous reported high performance liquid chromatography- ultraviolet (HPLC-UV) methods for the determination of these flavonoids have several disadvantages, including unsatisfactory separation times and not being well validated according to International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) standard guidelines. A rapid, specific reversed-phase HPLC method with isocratic elution of acetonitrile: isopropyl alcohol: 20 mM phosphate buffer (NaH2-PO4) (30:15:55, v/v) (pH 3.5) at a flow-rate of 1 ml/minute, a column temperature of 25_C, and ultraviolet (UV) detection at 340 nm was developed. The method was validated and applied for quantification of different types of O stamineus extracts and fractions. The method allowed simultaneous determination of 3’-hydroxy-5,6,7,4’- tetramethoxyflavone, sinensetin, and eupatorin in the concentration range of 0.03052 e250 mg/ml. The limits of detection and quantification, respectively, were 0.0076 and 0.061 mg/ml for 3’-hydroxy-5,6,7,4’-tetramethoxyflavone, 0.0153 and 0.122 mg/ml for sinensetin and 0.0305 and 0.122 mg/ml for eupatorin. The percent relative standard deviation (% RSD) values for intraday were 0.048e0.368, 0.025e0.135, and 0.05e0.476 for 3’-hydroxy-5,6,7,4’-tetramethoxyflavone, sinensetin, and eupatorin, respectively, and those for intraday precision were 0.333e1.688, 0.722e1.055, and 0.548e1.819, respectively. The accuracy for intraday were 91.25%e103.38%, 94.32%e109.56%, and 92.85%e109.70% for 30-hydroxy-5,6,7,40-tetramethoxyflavone, sinensetin, and eupatorin, respectively, and those for interday accuracy were 97.49%e103.92%, 103.58% e104.57%, and 103.9%e105.33%, respectively. The method was found to be simple, accurate and precise and is recommended for routine quality control analysis of O stamineus extract containing the three flavonoids as the principle components in the extract.
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