Acute toxicity of Stizophyllum perforatum leaves crude extract and determination of verbascoside by a validated HPLC-DAD method

Stizophyllum perforatum (Cham.) Miers (Bignoniaceae) crude extracts showed trypanocidal and leishmanicidal properties, additionally, verbascoside was the main active ingredient in the crude ethanol extract. We decided to quantify only verbascoside due to its biological properties and relative abundance in the crude extracts investigated during the study [1], [2], [3]. The aims of this study were to develop and validate a method for verbascoside determination in extracts of S. perforatum by HPLC-DAD using mangiferin as the internal standard, and to evaluate acute toxicity. The HPLC analysis was performed on a Luna ODS Phenomenex column, with a gradient mode, using water with acetic acid and acetonitrile. The method presented selectivity for verbascoside and mangiferin. The calibration curves were found to be linear in a concentration range of 0.8 – 60 µg/mL. In addition, this method provides recoveries between 102.82 and 111.04% and RSDs lower than 5.0%. In the acute toxicity test, the crude extract CE-2 did not cause deaths and significant behavioral changes in animals treated with a 2000 mg/kg dose. Thus, indicating a lethal dose of LD50 > 2000 mg/kg for CE-2. Additionally, macroscopic and histopathological analyzes confirmed that CE-2 did not induce significant toxicity in liver and kidney tissues. In CE-2 verbascoside concentration was 3.78 ± 0.14 µg/mL, 0.63% relatively to the dried extract. Thus, the developed method proved to be reliable and pertinent for quantification of verbascoside in S. perforatum samples, and regarding the acute toxicity study, the data suggested that CE-2 could be safe in a single administration.

Publikationsverlauf

Artikel online veröffentlicht:
12. Dezember 2022

• References

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