高效液相色谱-串联质谱法测定枇杷果膏中晚期糖基化终末产物和5-羟甲基糠醛

Determination of advanced glycation end products and 5-hydroxymethylfurfural in loquat fruit paste by high performance liquid chromatography-tandem mass spectrometry

中文关键词:  枇杷果膏，晚期糖基化终末产物，5-羟甲基糠醛，高效液相色谱质谱联用法，高效液相色谱法

英文关键词:loquat fruit paste  advanced glycation end products  5-hydroxymethylfurfural  high performance liquid chromatography-tandem mass spectrometry  high performance liquid chromatography

中文摘要:

      目的 建立高效液相色谱-串联质谱法(high performance liquid chromatography-tandem mass spectrometer, HPLC-MS/MS)检测果膏中晚期糖基化终末产物(advanced glycation end products, AGEs)和5-羟甲基糠醛(5-hydroxymethylfurfural, 5-HMF)的方法。方法 枇杷果膏以纯水浸提、超声处理后, 再经离心、固相萃取、氨水-甲醇溶液洗脱得到AGEs提取液, 以0.1%甲酸水和甲醇为流动相, 梯度洗脱, 采用多反应监测正离子源模式检测; 果膏经纯水浸提、乙酸乙酯提取、固相萃取、10%甲醇洗脱得到5-HMF提取液, 以体积比10%甲醇水为流动相, 在柱温30 ℃下检测。结果 3种目标物在浓度范围内均线性关系良好; 检出限和定量限均小于1 [AGEs/(ng/mL); 5-HMF/(μg/mL)], 可满足痕量物质的检测要求; 相对标准偏差(relative standard deviations, RSDs)均小于10%, 且在16 h样品待测时间范围内低于0.6%; 加标回收率均大于85%。结论 本研究优化的方法可满足定量分析的基本要求, 具有良好的准确性和稳定性, 为热加工果蔬产品质量与安全研究提供了检测手段。

英文摘要:

      Objective To establish a method for the determination of advanced glycation end products (AGEs) and 5-hydroxymethylfurfural (5-HMF) in loquat fruit paste by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Methods AGEs solution was obtained by extracting loquat fruit paste with pure water and treating with ultrasonic, centrifugation, solid-phase extraction and ammonia methanol solution elution, then separated by using gradient elution with 0.1% formic acid in water and methanol as the mobile phase. The detection method employed multi-reaction monitoring with positive ion. The 5-HMF solution was obtained by extracting the fruit paste in pure water, followed a liquid-liquid extraction with ethyl acetate, solid-phase extraction and 10% methanol elution. The extracts were analyzed at 30 ℃ with 10% methanol in water as mobile phase. Results It was found that the linearity of all the targets was good in the concentration range. The limits of detection and limits of quantification of each target substance were lower than 1 [AGEs/(ng/mL); 5-HMF/(μg/mL)], which could fulfill the requirements of detecting trace substances. The relative standard deviations (RSDs) of all the substances were less than 10%, and lower than 0.6% in the range of 16 hours of sample waiting time. The recoveries of all the substances were higher than 85%. Conclusion Results indicate that the method meet the basic requirement for quantitative analysis and has good accuracy and stability, which offers a detection means of studying on quality and safety of heat-processed fruit and vegetables.