Liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (QTOF/MS) assay for quantification of protodioscin in Brachiaria grasses.

This work aimed to develop and validate a method to detect and quantify protodioscin in Brachiaria grasses using ultraperformance liquid chromatography (UPLC) coupled to high-resolution quadrupole time-of-flight mass spectrometry. Samples were extracted by acetonitrile-water 50:50 v/v mixture and ultrasonication. The mobile phase consisted of 5 mM ammonium acetate in water-methanol and acetonitrile containing 0.1% formic acid. The parameters used to validate the method for determining protodioscin comprised determination of the selectivity, ionization suppression/enhancement (matrix effect), linearity of the calibration curve, the limit of detection (LOD), the lower limit of quantitation (LLOQ), and the precision and accuracy of the method. The LLOQ of protodioscin was determined as 0.1 µg mL-1, and the LOD was 0.03 µg mL-1. The developed method was applied for determining protodioscin levels in B. decumbens collected from three pastures where sheep showed clinical signs of photosensitization. The obtained values ranged from 0.71% to 1.12%. Thus, the developed method for determining protodioscin in Brachiaria grasses by LC coupled to high-resolution quadrupole time-of-flight mass spectrometry showed high accuracy, precision, and sensitivity.

Acute and sub-chronic toxicity of aqueous extracts of Chenopodium ambrosioides leaves in rats.

The present study aimed to evaluate the toxicity of aqueous extract of Chenopodium ambrosioides leaves. To measure acute toxicity, rats were administered 0, 0.3, 1.0, or 3.0 g/kg of aqueous extract from C. ambrosioides leaves by gavage. To analyze sub-chronic toxicity, rats were treated by oral gavage for 15 consecutive days with 0, 0.3, or 1.0 g/kg of extract of C. ambrosioides leaves. No animals from either trial exhibited any signs of toxicity. In the acute study, the highest dose of the extract led to an increase in the serum activities of alanine transaminase (ALT) and aspartate transaminase (AST) and a decrease in the serum levels of urea. In the sub-chronic test, rats treated with 1.0 g/kg for 15 days exhibited increased serum ALT activity and creatinine levels and mild cytoplasmic vacuolation of hepatocytes. The results indicate that aqueous extract from C. ambrosioides leaves produce slight hepatotoxic lesions in rats.