超高效液相色谱质谱联用技术在发酵乳有机酸分析中的应用  

Application of UPLC-MS in analysis of organic acids in fermented milk

作　　者：米智慧 陈永福[1] 张和平[1] 

机构地区：[1]内蒙古农业大学乳品生物技术与工程教育部重点实验室奶制品加工农业部重点实验室,呼和浩特010018

出　　处：《中国乳品工业》2017年第12期37-39,共3页China Dairy Industry

基　　金：国家自然科学基金(31571814);国家奶牛体系建设项目CARS-36)

摘　　要：利用超高效液相色谱-质谱联用(UPLC-MS)技术,建立了发酵乳中乙酸,乳酸,琥珀酸的检测方法。色谱条件为:采用ACQUITY UPlC HSS T3色谱柱进行分离,乙腈-0.1%甲酸/1 mmol/L甲酸铵水溶液体系作为流动相进行梯度洗脱,流速为0.25 mL/min,柱温为30℃,进样量5μL。质谱采集选择ESI-负离子监测模式进行扫描,优化锥孔电压和碰撞能量等质谱条件,离子源温度100℃,脱溶剂气温度450℃,脱溶剂气流量600 L/h。通过绘制有机酸的标准曲线,得到乙酸、琥珀酸、乳酸的线性范围分别是0~100,0~80,0~600μg/mL;最低检出限分别为0.04,0.0025,0.0025μg/mL;最低定量限分别为0.2,0.01,0.01μg/mL;相关系数范围R2为0.9951~0.9997,回收率范围为93.5%~106.07%。该方法准确、快速,可用于检测发酵乳中有机酸的含量。This study established a method for detecting the three kinds of short-chain fatty acids, acetic acid, lactic acid, and succinic acid of the fermented milk by ultra-performance liquid chromatography-mass spectrometry （UPLC-MS）. Chromatographic conditions were as fol- lows： the ACQUITY UPLC HSS T3 was used to separate the organic acids, acetonitrile solution-0.1% formic acid/1 mmol/L ammonium formate aqueous solution as mobile phase to gradient elute the sample, the flow rate was 0.25 mL/min, column temperature was set to 30 ℃, the injection volurrm was 5 μL. The ESI- negative mode was selected to monitor the changes,in addition,the cone voltage and collision en- ergy were optimized to obtain an appropriate method, the operating conditions were as follows：source temperature 100 ℃, desolvation tem- perature 450 ℃, desolvation gas flow 600 L/h, cone gas flow 50 L/h. By drawing the standard curve of organic acid, the linear range of acetic acid, lactic acid and succinic acid were ensured as follows：0~100,0-80,0-600 μg/mL respectively;the minimum detection limits were 0.04, 0.0025 and 0.0025 μg/mL;the minimum quantitative limits were 0.2,0.01,0.01 p,g/mL;the range of correlation coefficient Ik2 was between 0.9951-0.9997, and the recoveries varied between 81.3%~102.2%. The established method was proved to be accurate and fast for the determi- nation of organic acids in fermented milk.

关 键 词：超高效液相色谱-质谱联用仪 发酵乳 有机酸 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程]