Analysis of Prenylflavonoids from aerial parts of Epimedium grandiflorum and dietary supplements using HPTLC, UHPLC-PDA and UHPLC-QToF along with chemometric tools to differentiate Epimedium species

Authors: BAE, JI-YEONG - University Of Mississippi; AVULA, BHARATHI - University Of Mississippi; ZHAO, JIANPING - University Of Mississippi; RAMAN, VIJAYASANKAR - University Of Mississippi; WANG, YAN-HONG - University Of Mississippi; WANG, MEI - University Of Mississippi; ZULFIQAR, FAZILA - University Of Mississippi; FENG, WEI - National Institutes For Food And Drug Control; PARK, JONG HEE - Pusan National University; ABE, NAOHITO - Gifu University; ALI, ZULFIQAR - University Of Mississippi; KHAN, IKHLAS - University Of Mississippi

Submitted to: Journal of Pharmaceutical and Biomedical Analysis
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/27/2019
Publication Date: 8/28/2019
Citation: Bae, J., Avula, B., Zhao, J., Raman, V., Wang, Y., Wang, M., Zulfiqar, F., Feng, W., Park, J., Abe, N., Ali, Z., Khan, I.A. 2019. Analysis of Prenylflavonoids from aerial parts of Epimedium grandiflorum and dietary supplements using HPTLC, UHPLC-PDA and UHPLC-QToF along with chemometric tools to differentiate Epimedium species. Journal of Pharmaceutical and Biomedical Analysis. 177:112843. https://doi.org/10.1016/j.jpba.2019.112843.
DOI: https://doi.org/10.1016/j.jpba.2019.112843

Interpretive Summary: • Epimedium has been traditionally used in China, Japan and Korea to treat osteroporosis, sexual dysfunction and cardiovascular disease. • There is no reports available on the chemical analysis of E. grandiflorum. • The purpose of this study is to evaluate the chemical composition of E. grandiflorum and compare with other species of Epimedium (E. brevicornu , E. sagittatum and E. koreanum). • A simple and rapid UHPLC-PDA validated method was developed for determination of fifteen prenylflavonoids from E. grandiflorum. • The developed method was validated for all parameters tested. • Chemometric analysis including PCA was used for the differentiation of three Epimedium species (E. grandiflorum, E. brevicornu and E. sagittatum). • A simple and fast HPTLC method was also developed for the chemical fingerprint analysis of nine prenylflavonoids, are useful in assessing the quality of dietary supplements claiming to contain E. grandiflorum.

Technical Abstract: An Ultra-High Performance Liquid Chromatography (UHPLC) method was developed for the determination of 15 prenylflavonoids from aerial parts of Epimedium grandiflorum and related species (Berberidaceae). The separation was achieved within 8 minutes by using C-18 column material, a water/acetonitrile mobile phase, both containing 0.1% formic acid gradient system and a temperature of 40°C. The method was validated for linearity, limits of detection (LOD) and limits of quantification (LOQ) and repeatability. The limits of detection were found to be in the range from 0.1-0.5 µg/mL. The wavelength used for quantification with the photodiode array detector was 269 nm. The total content of 15 prenylflavonoids were 9.1-20.6 mg/g for E. grandiflorum (except for sample #2899 and #20862), 5.6-35.4 mg/g for E. brevicornu and 10.8-30.5 mg/g for E. sagittatum. Twenty dietary supplements contained compounds in the range from 0.1-81.7 mg/day. The developed method is simple, economic, rapid and especially suitable for quality control analysis of E. grandiflorum. Liquid chromatography quadrupole time of flight-mass spectrometry (LC-QToF-MS) with electrospray ionization (ESI) method is described for the identification and confirmation of compounds in plant samples and dietary supplements. This technique is also used for chemical profiling of Epimedium samples. This method involved the use of [M+H]+ and [M+Na]+ ions in the positive ion mode and [M-H]- and [M+HCOO]- ions in the negative ion mode with extracted ion monitoring (EIM). The molecular formula determination and tentative identification of compounds was based on accurate mass measurement and fragments using MS-MS accurate mass spectra with collision induced dissociation (CID) of precursor ions. Chemometric analytical tool for visualizing plant and commercial samples quality was used for discriminating between Epimedium species and dietary supplements with regards to the relative content or presence of components. A High-Performance Thin Layer Chromatography (HPTLC) method was also developed for the fast chemical fingerprint analysis of Epimedium species.