Simultaneous Quantification of Berberine and Curcumin in Combined Extract Form and Polyherbal Marketed Formulation

An RP-HPLC analytical method for the estimation of both herbal markers (berberine and curcumin) was developed by Waters 515 Series pumps connected with an auto sampler, injector with a 20µl fixed loop and a 2998 PDA UV–vis. detector.

Separation was performed on an Agilent C-18(2) column (particle size 5µm; 250mm×4.5mm SN-usjab01584). Chromatographic data were recorded and processed by means of an EMPOWER-2 tool and photo diode array (PDA) detector. The mobile phase used was ACN: 20mM KH₂PO₄ buffer solution (PBS) (adjusted to pH 3.0 with phosphoric acid, H₃PO₄) (35:65 v/v) of 1.0 mL/min. The elution was measured at 272 nm.

The Rt was 3.07 and 5.53 min for berberine and curcumin respectively. The linearity range was from 150-4800 ng/mL and 900-28800 ng/mL for berberine and curcumin respectively. The LOD and LOQ were found to 50 ng/mL and 150 ng/mL for berberine and 300 ng/mL and 900 ng/mL for curcumin. Moreover, reproducibility and repeatability were found within the range i.e., % RSD ≤ 2% for both drugs. The accuracy of berberine and curcumin was 98.15% w/w and 98.57% w/w respectively. The %age purity of marketed herbal formulation was 94.61% w/w and 95.94% w/w for berberine and curcumin respectively.