高效液相色谱法测定钙片中酪蛋白磷酸肽的含量

Determination of Casein Phosphopeptides in Calcium Tablets by High Performance Liquid Chromatography

摘要:

采用高效液相色谱（HPLC）法测定了5种钙片中酪蛋白磷酸肽（CPP）的含量。在流动相为水+1‰ TFA（水相）和乙腈+1‰ TFA（有机相），检测波长215 nm，流速1 mL/min，进样体积10 μL下，通过HPLC色谱图分析，优选了梯度洗脱条件为有机相5%~35%，45 min。在此条件下，CPP分离效果良好，回收率介于96.75%~104.07%，比较接近真实值，说明这5种钙片中CPP添加效果良好。同时，结果也表明，HPLC法处理快速简便，准确度和灵敏度较高，检测限为6.81 μg/mL，可作为检测钙片中CPP含量的优良方法。另外，为了将HPLC法与其他检测方法进行比较，还采用应用较普遍的钡-乙醇沉淀法对相同的5种钙片中的CPP含量进行了测定，结果表明，5种钙片样品中的CPP回收率均超过了220%，偏差较大，因而此法不宜作为钙片中CPP的检测方法。

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.