Development and validation of HPLC method for vicenin-1 isolated from fenugreek seeds in rat plasma: application to pharmacokinetic,tissue distribution and excretion studies

Context: Vicenin-1, a flavonol glycoside, has potent platelet aggregation inhibition, antioxidant, radioprotectants and anti-inflammatory activities.

Objective: To establish a rapid, simple, precise and sensitive high-performance liquid chromatography (HPLC) method for determination of vicenin-1 in rat plasma, and to investigate the pharmacokinetics, tissue distribution and excretion after a single 60?mg/kg oral dose in rats.

Materials and methods: Vicenin-1 was extracted by solid–liquid extraction through Tulsicon^® ADS-400 (0.40–1.2?mm). Chromatographic separation was achieved by HPLC with a C₁₈ column with a mobile phase composed of water and acetonitrile (75:25 v/v) and a flow rate of 1?mL/min along with UV detection at 210?nm.

Results: Good linearity of calibration curve was found between 10.5 and 100.5?μg/mL (R^2?=^?0.995) for plasma and tissue, whereas 2.5–500?μg/mL (R^2?=^?0.999) for the urine and stool samples. The extraction recoveries were 98.51–99.58% for vicenin-1 in plasma, whereas intra-day and inter-day precision were validated by relative standard deviation (%RSD), that came in the ranges of 1.16–1.79% and 1.28–1.73%, respectively. The pharmacokinetics results showed C_max (7.039?μg/mL) and T_max (2?h) after oral administration of vicenin-1. Tissue distribution study showed that the highest concentration of vicenin-1 was achieved in the liver followed by the lung. Approximately 24.2% of its administered dose was excreted via urinary excretion route.

Conclusion: The first-pass metabolism, poor solubility and presence of reducing sugar moiety in vicenin-1 may decrease its bioavailability. The developed method is sensitive, specific and was successfully applied to the pharmacokinetics, tissue distribution and excretion studies of vicenin-1 in rats.