کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1213104 | 1494109 | 2013 | 10 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Development and validation of a rapid ultra-high performance liquid chromatography diode array detector method for Vitex agnus-castus Development and validation of a rapid ultra-high performance liquid chromatography diode array detector method for Vitex agnus-castus](/preview/png/1213104.png)
A rapid ultra-high performance liquid chromatography diode array detector (UHPLC-DAD) method was developed and validated for the simultaneous determination of all classes of non-volatile phytochemicals (iridoids, flavonoids and diterpenes) in Vitex agnus-castus (Lamiaceae) fruits, a traditional medicinal plant used against premenstrual symptoms (PMS) and other disorders. Seven marker compounds, 3,4-dihydroxybenzoic acid, p-hydroxybenzoic acid, agnuside, 5-hydroxykaempferol-3,6,7,4′-tetramethylether, 1,2-dibenzoic acid glucose, methoxy-vitexilactone, and vitetrifolin D were isolated from the methanol extract of V. agnus-castus to be used as reference substances. Chromatographic separation was performed on a Zorbax Eclipse XDB-C18 (50 mm × 2.1 mm) UHPLC column with 1.8 μm particle size, within 20 min. A solvent gradient from 0.5% acetic acid to acetonitrile at a flow rate of 0.6 mL/min was used as mobile phase. Analyte detection and quantification was realized at 210 nm and 260 nm. The UHPLC-DAD assay was validated for the quantitative analysis of agnuside, isovitexin, casticin, 5-hydroxykaempferol-3,6,7,4′-tetramethylether and vitetrifolin D. It was found to be specific, accurate, precise, and reproducible for the quantification of these compound within a concentration range of 0.7–500.0 μg/mL for casticin and 5-hydroxykaempferol-3,6,7,4′-tetramethylether, 1.4–1000.0 μg/mL for isovitexin and agnuside, and 12.4–1000.0 μg/mL for vitetrifolin D. Intra- and inter-day variations showed relative standard deviations (RSD) of less than 3.9% and 6.4%, respectively. Tentatively assignment of 62 chromatographic features found in the UHPLC-DAD assay was carried out by coupling the UHPLC instrument to a quadrupole time-of-flight mass spectrometer via an electrospray ionization interface (ESI-QTOF-MS) operated in positive and negative ion mode. By using the established quantitative UHPLC-DAD assay to asses agnuside, isovitexin, casticin, 5-hydroxykaempferol-3,6,7,4′-tetramethylether and vitetrifolin D in V. agnus-castus derived preparations as extracts, tinctures and tablets, the applicability of the developed assay to phytopharmaceuticals was successfully proven.
► A rapid UHPLC-DAD assay to monitor Vitex agnus-castus was developed.
► The assay covers all major classes of non-volatile V. agnus-castus phyto-chemicals.
► Based on UHPLC-DAD-QTOF-MS experiments 62 chromatographic peaks of V. agnus-castus extract could be characterized.
► The assay was validated and can be applied to fruits, tablets, and extracts.
Journal: Journal of Chromatography B - Volume 927, 15 May 2013, Pages 181–190