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Objective: The objective of this research was to study the retention behavior of β-carotene under isocratic elution by reversed-phase high performance liquid chromatography (RP-HLPC) for describing its elution behavior. Method: The reversed-phase chromatographic elution systems were performed using binary mobile phase comprising 2-propanol in methanol at different volume fractions (ψ) under the designed temperatures at 25 and 35 oC. In this study, the obtained chromatographic parameters, i.e. retention factor (k) and reciprocal retention factor (1/k) were calculated to provide the plots for describing the elution behavior. Results: From this study, the exponential dependency of k on ψ, and linear dependency of 1/k on ψ were plotted. From linear plot, the derived chromatographic parameters, i.e. slope (S) and extrapolated y-intercept (1/ko) were also considered. The larger value of correlation coefficient (r2) from the linear plot at 35 oC (r2 = 0.9922) has been obtained with the comparison of r2 at 25 oC (r2 = 0.9783). These findings thus have recommended that the elution behavior of β-carotene using 2-propanol in methanol at ψ = 0.25% was able to elute β-carotene successfully with the faster elution at 35 oC evidently.Conclusion: The elution behavior of β-carotene under its partition between the binary mobile phases and immobilized n-alkyl chains in stationary phase was depended on the polarity of mobile phase as a function of temperature used. These results will be useful for predicting and controlling the elution behavior of β-carotene under isocratic RP-HPLC system.

international journal of pharma medicine and biological sciences

Effects of Mobile Phase Composition as a Function of Temperature on Isocratic Elution Behavior of β-carotene in Revered-Phase Liquid Chromatographic Systems