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Anubala S.,Natural Product Chemistry Division | Sekar R.,Analytical Chemistry Division | Narayana P.S.,Analytical Chemistry Division | Nagaiah K.,Indian Institute of Chemical Technology
Journal of Planar Chromatography - Modern TLC | Year: 2015

Piperine (PIN), a bioenhancer, is co-formulated with rifampicin (RFN) and isoniazid (INZ) to enhance the bioavailability of RFN. RFN and INZ are the drugs of first choice for the treatment of tuberculosis and recommended by the World Health Organization (WHO). A new high-performance thin-layer chromatographic (HPTLC) method was optimized for the separation and determination of RFN, INZ, and PIN in the bioenhanced pharmaceutical formulation. Separation was performed on a precoated silica gel 60 F254 plate, and an acceptable separation was achieved using a mobile phase comprising toluene-ethyl acetate-methanol (7:3:3, v/v); detection was performed at 300 nm. The proposed method was validated for specificity, accuracy, repeatability, limit of detection, limit of quantification, stability, and robustness. The correlation coefficient (Rsup) values for RFN, INZ, and PIN were 0.9997, 0.9995, and 0.9987, respectively. The spots were identified by matching with the RF values and absorbance spectrum with authentic compounds. The mean recovery (%) values for RFN, INZ, and PIN were 99.3, 99.8, and 99.2%, respectively. The results obtained with the proposed method have been compared with those previously reported for high-performance liquid chromatographic (HPLC) methods. The major degradation products of RFN: 3-formyl rifamycin, isonicotinyl hydrazone (INH), and rifampicin quinone did not interfere with the determination of active pharmaceutical ingredients (APIs). The proposed method is also suitable for the detection of some major RFN-related impurities in pharmaceutical dosage form. © 2015 Akadémiai Kiadó, Budapest.

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