Development of a RP-HPLC method for simultaneous determination of lactic acid，fumaric acid, 5-HMF and patulin in apple juice

同时定量检测苹果汁中乳酸、富马酸、5-HMF和展青霉素，建立一种基于多通道紫外检测器的反相高效液相色谱（RP-HPLC）分离方法。采用C18色谱柱（250 mm×4.6 mm ID,5.0 μm），以pH为2.5的磷酸水溶液和乙腈按95:5混合为流动相，在流速1.00 ml*min-1，温度30℃下进行乳酸、富马酸、5-HMF和展青霉素的色谱分析。前10 min，两种有机酸在210 nm下被分离和检测；后10 min，5-HMF和展青霉素于276 nm下被分离和检测。检测限分别为: 乳酸0.1790 mg*L-1、富马酸0.0032 mg*L-1、5-HMF 0.0057 mg*L-1和展青霉素0.0089 mg*L-1，保留时间、峰高和峰面积的重现性（n=3）变异系数（CV）均小于1.0%。保留时间的日内重现性（n=3）和日间精密度（n=10）变异系数（CV）分别小于0.09%和0.5%。平均回收率为95.64%～99.17%。该方法可应用于同时定量检测苹果浓缩汁中乳酸、富马酸、5-HMF和展青霉素。 A reversed-phase high-performance liquid chromatography (RP-HPLC) separation method with ultraviolet detector was developed to determine the lactic acid, fumaric acid, 5-hydroxymethylfurfural (5-HMF) and patulin in apple juice simultaneously. The above four organics were determined by RP-HPLC using a C18 (250 mm×4.6 mm ID, 5.0 μm) column and the mixture phosphoric acid solution (pH 2.5)-acetonitrile (95:5, v/v) as the mobile phase with a flow rate of 1.00 ml*min-1 at 30℃. Lactic acid and fumaric acid were eluted and detected in the first 10 min at 210 nm. 5-H