高效液相色谱法测定钙片中酪蛋白磷酸肽含量.pdfVIP

现代食品科技 Modern Food Science and Technology 2014, Vol.30, No.7 258 高效液相色谱法测定钙片中酪蛋白磷酸肽的含量 张帅 1，2 ，刘飞 3 ，罗珍 3 ，郭斌 3 ，彭维 4 ，曹庸 2 （1.肇庆学院化学化工学院，广东肇庆 526061）（2.华南农业大学食品学院，广东广州 510642） （3.无限极（中国）有限公司，广东广州 510665）（4.广州医科大学公卫学院，广东广州 510182） 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。 关键词：酪蛋白磷酸肽；高效液相色谱；钙片；梯度洗脱；回收率 文章篇号：1673-9078(2014)7-258-262 Determination of Casein Phosphopeptides in Calcium Tablets by High Performance Liquid Chromatography ZHANG Shuai1,2, LIU Fei3, LUO Zhen3, GUO Bin3, PENG Wei4, CAO Yong2 (1.College of Chemistry and Chemical Engineering, Zhaoqing University, Zhaoqing 526061, China) (2.College of Food Science, South China Agricultural University, Guangzhou 510642, China) (3.Infinitus (China) Co., Ltd., Guangzhou 510665, China) (4.Institute of Public Health, Guangzhou Medical University, Guangzhou 510182, China) 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 excell