A validated HPTLC method for determination of trans-caryophyllene from polyherbal formulations

Formulations of traditional medicines are usually made up of a complex mixture of herbs. However, effective quality control methods in order to select materials of the right quality are lacking. 'Amukkara choornam' is a polyherbal Siddha formulation used for gastritis, spleen enlargement, leucorrhoea, hiccups, anaemia, tuberculosis and kappa diseases. Trans-caryophyllene is an important constituent present in the ingredients of this formulation. In a literature survey, it was found that there is no such method for the quantification of trans-caryophyllene except gas chromatography or gas chromatography-mass spectroscopy (GC-MS). So, a high performance thin layer chromatography (HPTLC) method was developed and validated for the quantification of trans-caryophyllene in amukkara choornam. Pre-coated silica gel 60F-254 plates (10 × 10 cm²) were used for the analysis. The solvent system consisted of toluene-ethyl acetatate (9 : 3, v/v), and trans-caryophyllene was detected at 260 nm. The developed method was validated for linearity (R² = 0.9996 ± 0.0034), limit of detection (LOD) (0.101 ng), limit of quantification (LOQ) (0.639 ng), accuracy (% recovery = 97.19 ± 1.204), and precision (CV < 5%, for both intra-day and inter-day precisions). The levels of trans-caryophyllene were found to be 3.5-4.10 µg per gram of herbal products.