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Determination of Biflavonoids in Four Mexican Species of Selaginella by HPLC

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The genus Selaginella comprises several species, some of which grow on the American continent. Among these species, S. nothohybrida Valdespino, S. lepidophylla Hook et Grev, S. pallescens (Presl) Spring, and S. reflexa Underw. are found in several states of the Mexican Republic. The aforementioned three species are used medicinally, typically to treat renal disorders. This paper describes the development of an HPLC-UV method for the determination of amentoflavone (1), robustaflavone (2), and (2S)-2,3-dihydrorobustaflavone (3) as major components of the above-mentioned species of Selaginella. Components 1, 2, and 3 were quantitatively determined using an XBridge Waters C18 5 μm column, with a gradient system consisting of mixtures of acetonitrile and water with 0.4% acetic acid. The flow rate was 0.4 mL/min, with UV detection at 367 nm. LOD and LOQ values were in the range of 0.025 to 0.216 μg/mL. Compounds 1, 2, and 3 showed good linearity in the 1.2 to 18 μg/mL range; recovery was within 98.2 and 101.9% for all three cases. Compounds 1 and 2 were detected in all eight samples; their concentration ranged from 0.35 to 1.79 mg/g of plant. Thus, compounds 1 and 2 could be used as markers for S. nothohybrida, S. lepidophylla, S. pallescens, and S. reflexa. In addition, trehalose was detected in all samples as two peaks at 1.5 and 2.0 min. The HPLC method described here was shown to be reliable, reproducible, and accurate and can be used for QC of Selaginella medicinal materials.

Document Type: Research Article


Publication date: July 1, 2013

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  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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