BIOANALYTICAL METHOD VALIDATION FOR DETERMINATION OF ROSMARINIC ACID IN SIMULATED BIOLOGICAL MEDIA USING HPLC

Objective: This study aims to develop and validate a simple, rapid accurate HPLC method for the determination of rosmarinic acid (RA), an active marker Thunbergia laurifolia (TL) tea, in Hank's Balanced Salt Solution (HBSS) using HPLC.
 Methods: The separation was performed Xterra® C18 reversed-phase column (150 3.9 mm, 5 µm). mobile phase consisted 0.5% (v/v) glacial acetic (A) acetonitrile (B). flow rate 1 ml/min detection carried out at 330 nm with UV-visible spectrophotometer. validated according US FDA guidance on bioanalytical validation 2018.
 Results: successfully range 50-500 ng/ml RA. Intra-and inter-day precision ranged from 1.6 2.1% 1.4 4.5%, respectively. relative errors (% bias) were less than 7.6 3.6%, In addition, it found that stability RA HBSS could be dependent pH temperature.
 Conclusion: developed met requirements further applied permeability vitro Caco-2 cell monolayer model.