Determination of Casein Phosphopeptides in Calcium Tablets by High Performance Liquid Chromatography
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Abstract:
The contents of casein phosphopeptides (CPP) from five kinds of calcium tablet were determined by high performance liquid chromatography (HPLC) method. Mobile phase, detection wavelength, velocity and injection volume were the mixture of 1‰ TFA aqueous solution and 1‰ TFA acetonitrile solution, 215 nm, 1 mL/min, and 10 μL, respectively. The optimal condition of gradient elution was obtained by HPLC chromatogram analysis as organic phase of 5%~35% and elution for 45 min. Under the optimal conditions, CPP can be well seperated, and its recovery was between 96.75%~104.07%, which was close to the true value. This method was proved to be simple with high accuracy and sensitivity, and its detection limit was 6.81 μg/mL, which could be regarded as an excellent method to determine CPP in calcium tablets. In order to compare this HPLC method with the other methods, Barium-ethanol precipitation was used to determine the CPP contents in the same five calcium tables. Result showed that the recoveries of CPP in five samples were all more than 220%, which was inappropriate to determine the content of CPP in calcium tablets for the great deviation.
