柱前衍生高效液相色谱法检测乳制品中羟脯氨酸

采用高效液相色谱法(HPLC)检测乳制品中羟脯氨酸(Hyp)。以RP-HPLC为分离模式,邻硝基苯磺酰氯为衍生剂,选用Hedera ODS-2色谱柱(250mm×4.6 mm,5μm),15 mmol/L磷酸盐缓冲液(pH 5.8)和乙腈为流动相梯度洗脱,检测波长为230nm,建立了在多种氨基酸中分离出羟脯氨酸的方法。Hyp在9.70×10-3～4.96 mmol/L范围内峰面积与浓度线性良好,相关系数为0.9999,加标回收率范围95.5%～102.7%,峰面积的相对标准偏差为1.5%。本法适用于检测乳制品中的Hyp,具有准确度高,操作简单等优点。

Determination of hydroxyproline in dairy products by high performance liquid chromatography with precolumn derivatization

A method was developed for the determination of hydroxyproline by reversed phase high performance liquid chromatography.Under the optimal conditions,a Hedera ODS-2 column(250 mm ×4.6 mm,5 μm) was used,and the detection wavelength was 230 nm.Flow rate was 1.0 mL/min;Mobile phase A: 15 mmol/L sodium dihydrogen phoephate(pH 5.8),Mobile phase B: acetonitrile;the gradient protocol was applied.The correlation coefficient(r) of regression equation was 0.9999,and the recoveries were from 95.5% to 102.7%.The relative standard deviation was 1.5%.The method is accurate,easy and so on,and it can be applicable to the detection of hydroxyproline in dariy products.