Quantitative Estimation of Piperine in Ayurvedic Formulations by HPTLC

Churna are important group of formulation used by Ayurvedic physicians to treat various types of diasease. Trikatu and Pimpali churna, as per Ayurvedic literature is used for the treatment of resperatory disorders. In the present study, an attempt has been made to develop a HPTLC method of quantitative estimation of marker compound, piperine in laboratory prepared authentic formulation and a marketed formulation of Trikatu and Pimpali churna. The stationary phase used was precoated silica gel G₆₀ F₂₅₄ plates. The mobile phase containing n-Hexane-ethyl acetate, (5:5 v/v), was used to separate the spot of Piperine. Plates were developed to a distance of 8 cm at room temperature. Spectrodensitometric scanning was performed by TLC scanner III (CAMAG) in absorbance mode at the wavelength of 339nm. The R_f values of piperine was 0.44± 0.02. The method was validated in terms of Linearity, Accuracy and Precision. The linearity curve found to be linear in between 200-900 ng/spot. The limit of Detection (LOD) and limit of Quantification (LOQ) were found to be 1 ng and 3 ng respectively. The mean results from % Recovery was found to be, laboratory formulation contain 85.13% and 83.47% while the commercial formulation shows 96.65%, and 90.32% respectively. The proposed method can be used to determine Piperine from Ayurvedic formulations.