RESUMO
Objective: A simple, accurate, and rapid high-performance thin-layer chromatographic (HPTLC) method for simultaneous quantification of the two biologically active flavonoidal compounds, gallic acid and quercetin, in Hygrophila auriculata (K. Schum) Heine (HA) has been established and validated. Methods: Chromatography was performed on aluminium foil-backed silica gel 60 F254 HPTLC plates with the binary mobile phase toluene: ethyl acetate: formic acid (5:4:1, v/v/v). Ultraviolet detection was performed densitometrically at the maximum absorbance wavelength, 270nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with ICH guidelines. Results: The system was found to give compact spots for gallic acid (GA) and quercetin (QE) (Rf value of 0.31 and 0.50, respectively). The limit of detection (23 and 41 ng band-1) limit of quantification (69 and 123 ng band-1), recovery (99.4-99.9 and 98.7-99.4%), and precision (i.e ≤1.98 and 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for GA and QE were 100-1000 (r 2= 0.9991) and 150-900 ng band-1 (r2= 0.9956) and the contents estimated as 0.28±0.01% and 0.41±0.01% w/w respectively.Conclusions:This simple, precise and accurate method gave good resolution from other constituents present in the extract. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing Hygrophila auriculata (K. Schum) Heine.
RESUMO
Objective: To analyse the two marker compounds lupeol (LP) and stigmasterol (ST) from methanolic extract of Hygrophila auriculata (H. auriculata). Methods: Separation was achieved on aluminium plates precoated with silica gel 60F254 with toluene-methanol-formic acid (7.0:2.7: 0.3 v/v/v) as mobile phase. Results: Densitometric analysis was performed at 530 nm in the reflectance mode. Compact bands for LP and ST were obtained at RF 0.52 ± 0.02 and 0.28 ± 0.05. Linearity (r2=0.998 5 and 0.993 7), limit of detection (45 and 18 ng/band) limit of quantification (135 and 54 ng/band), recovery (98.2%-99.7% and 97.2%-99.6%), and precision (<2.18 and 1.91) were satisfactory for LP and ST respectively. Linearity range for LP and ST were 100-1 000 and 50-500 ng/band and the contents estimated as (0.19±0.1)% and (0.47±0.1)% w/w respectively. Conclusion:The method demonstrated efficient analysis testing of LP and ST in samples; therefore it can be used for routine analysis.