Development of HPTLC method for determination of cefpodoxime proxetil and ambroxol hydrochloride in human plasma by liquid–liquid extraction
Aim: To develop a simple, sensitive, rapid, and economic high performance thin layer chromatographic method for determination of cefpodoxime proxetil (CEFPO) and ambroxol hydrochloride (AMBRO) in human plasma by liquid–liquid extraction using paracetamol as an internal standard. Materials and Methods: The plasma sample was extracted by a mixture of methanol and acetonitrile. A concentration range from 500 to 3500 ng/spot for CEFPO and 1000 to 7000 ng/spot for AMBRO were used for the calibration curve, respectively. This recovery was found to be 74.40 and 94.50 for CEFPO and AMBRO, respectively. The mobile phase used consists of chloroform: methanol (9:1v/v). Densitometric analysis was carried out at a wavelength of 240 nm. Results: The Rf values for CEFPO, AMBRO, and paracetamol were found to be 0.69 ± 0.005, 0.49 ± 0.0057, and 0.31 ± 0.0054, respectively. The stability of CEFPO and AMBRO in plasma was confirmed during three freeze–thaw cycles (-20°C), on bench during 24 h and post preparative during 48 h. Conclusion: The proposed method was validated statistically and by performing a recovery study for determination of CEFPO and AMBRO in human plasma.