Development and validation of a method for densitometric analysis of berberine in herbal extract and polyherbal...

To develop a new high-performance thin-layer chromatography (HPTLC) method for the quantification of berberine in herbal extract and pharmaceutical dosage form.

The HPTLC was performed on aluminium foil plates coated with 200 μm silica gel 60F(254). Linear ascending development with toluene:ethyl acetate:formic acid:methanol 9:9:3:1 (v/v/v/v) was performed at room temperature (25 ± 2°C) in a twin-trough glass chamber saturated with mobile-phase vapour. Compact bands (R(F) 0.58 ± 0.02) were obtained for berberine. Spectrodensitometric scanning was performed in fluorescence mode at 350 nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with International Conference on Harmonization guidelines.

Linear regression analysis of the calibration plots showed a good linear relationship (r(2) = 0.9996 ± 0.0001) between peak area and concentration in the range 10-100 ng/band, respectively. The limits of detection and quantification were 2.8 and 9.3 ng/band. The recovery of the method was 98.5-100.6%.

The above method was a rapid and cost-effective quality-control tool for routine analysis of berberine in herbal extracts and in pharmaceutical dosage form.