RP-HPLC法同时检测苹果汁中乳酸、富马酸、5-HMF和展青霉素的条件优化

Simultaneous Determination of Lactic Acid Fumaric Acid,5-HMF and Patulin in Apple Juice by RP-HPLC

建立了一种结合紫外检测器同时检测果汁中的乳酸、富马酸、5-羟甲基糠醛(5-hydroxymethylfurfural,5-HMF)和展青霉素的反相高效液相色谱(reversed-phase high-performance liquid chromatography,RP-HPLC)分离方法。考察了柱温、pH值、流速以及乙腈浓度对各组分保留时间和峰高的影响,最终采用pH为2.5的磷酸水溶液和乙腈的混合液作用于反相C18柱,以进行乳酸、富马酸、5-HMF和展青霉素的色谱分析。在流速为1.00ml/min,温度为30℃的整个分析过程中,流动相为含有5%乙腈的磷酸水溶液。前10min内,两种有机酸被在210nm处分离和检测。而5-HMF和展青霉素则被接下来的10min内于276nm处分离和检测。89.96%到102%的平均回收率证实了方法的准确度。该方法被成功地用于同时定量苹果浓缩汁中的乳酸、富马酸、5-HMF和展青霉素。

A reversed-phase high-performance liquid chromatography （RP-HPLC） separation method with ultraviolet detection was developed for the simultaneous determination of lactic acid, fumaric acid, 5-hydroxymethylfuffural （5-HMF） and patulin in apple juices. The chromatographic analysis of lactic acid, fumaric acid, 5-hydroxymethylfurfural and patulin was carried out after their elution with phosphoric acid solution （pH 2.5） and acetonitrile from RP-C18 stationary phase, and the effects of temperature, pH value, flow rate and concentration of acetonitrile on retention time and peek height were studied. The mobile phase employed was phosphoric acid solution with 5% acetonitrile working at flow rate of 1.00 ml/min at 30℃ for the whole run. Two organic acid acids were eluted within 10 min and were detected at 210 nm .The other two compositions, including 5- HMF and patulin, were eluted within the following 10 min and were detected at 276 nm. The accuracy of the method is confirmed with an average recovery range between 89.96 and 102%. The method is successfully used to assay lactic acid, fumaric acid, 5-HMF and patulin simultaneously in apple juice.