Evaluation of marketed herbal medicines for the simultaneous estimation of steroidal adulterants using FTIR and RP-HPLC-UV

The herbal products are considered safe due to their natural origin; however there are reports about their adulteration with synthetic steroids. In this particular study identification and quantification of selected steroidal adulterants including prednisolone, betamethasone, dexamethasone, triamcinolone acetonide and testosterone in herbal products have been carried out using FTIR and HPLC. The samples were extracted by methanol, and quantified by high-performance liquid chromatography (HPLC). The chromatographic separation was achieved on stationary phase Supelco R100–10 C18 ODS (5 µm × 250 × 4.6 mm) using methanol:water (60:40 v/v) as mobile phase (pH was adjusted to 4.5 with ortho-phosphoric acid) at a flow rate of 1.1 ml/min with a detection wavelength of 242 nm. The developed procedure was successfully applied to spiked plasma and herbal medicines samples. The LODs were 0.2, 0.02, 0.02, 0.06, 0.144 µg/ml, while LOQ were 0.6, 0.06, 0.07, 0.192 and 0.438 µg/ml for testosterone, prednisolone, triamcinolone, dexamethasone and betamethasone, respectively. The developed method was validated according to ICH guidelines. The regression coefficients of the analytes were 0.999 for standard solutions while for spiked plasma samples these were from 0.998 to 0.999. The proposed method is the first one for the simultaneous evaluation of the mentioned steroids. The method is simple, sensitive, and inexpensive that can be used for the studied steroids in dosage forms and body fluids.