CHEMOMETRIC ASSISTED RP-HPLC METHOD DEVELOPMENT & VALIDATION OF CYPROHEPTADINE HCL AND TRICHOLINE CITRATE

Abstract

Special, effective high pressure liquid chromatography method has been developed for the simultaneous
quantification of Cyproheptadine and Tricholine Citrate. By using Waters HPLC e-2695 quaternary pump with a
PDA detector of 2998 instrument the chromatographic separation of Cyproheptadine and Tricholine Citrate was
achieved on the column of Luna C18 (250x4.6mm, 5 µ) using an isocratic elution with a buffer containing water and
acetonitrile at a rate of 90:10 as a mobile phase with a flow rate of 1 ml/min at ambient temperature. A detector
wavelength of 245 nm utilizing the PDA detector was given in the instrumental settings. The linearity was studied
between the concentration range of 0.5-3 µg/ml of Cyproheptadine and 68.75-412.5µg/ml of Tricholine Citrate was
injected. The plotted calibration curves were linear with a regression coefficient of R2> 0.999, indicates that the
linearity was within the limit. As a part of method validation the parameters like specificity, linearity, accuracy,
ruggedness, and robustness were determined and the results were found to be within the allowable limit. The method
developed was found to be applicable to routine analysis and to be used for the measurement of both active
pharmaceutical ingredients (i.e., Cyproheptadine and Tricholine Citrate). Validation of the proposed method was
carried out according to an International Conference on Harmonization (ICH) guidelines. Since, there is no HPLC
method reported in the literature for the estimation of Cyproheptadine and Tricholine Citrate, there is a need to
develop quantitative methods under different conditions to achieve improvement in specificity, selectivity etc.