超高效液相色谱-质谱法测定母乳中12种低聚糖

Determination of 12 Oligosaccharides in Human Milk by Ultra-Performance Liquid Chromatography-Mass Spectrometry

建立母乳中12种低聚糖超高液相色谱-串联三重四极杆质谱的定性及定量测定方法。母乳样品脱脂除蛋白后经液相分离,通过质谱检测,最优梯度洗脱条件为:流动相A为10 mmol/L的甲酸铵溶液,流动相B为乙腈;流速0.3 m L/min;柱温50℃;0~10 min,95%~75%B;10~15 min,75%B;15~20 min,75%~65%B;20~21 min,65%~10%B;21~24 min,10%B;24~25 min,10%~95%B;25~35 min,95%B。12种低聚糖标准品标准曲线在设定质量浓度范围内线性良好(R2>0.98),加标回收率在80.00%~120.00%之间,重复性和仪器精密度(均用相对标准偏差表示)均小于10%。

A qualitative and quantitative method for the determination of 12 oligosaccharides in human milk was established. The conditions for gradient elution were set as follows： mobile phase A, 10 mmol/L ammonium formate solution; mobile phase B, acetonitrile; flow rate, 0.3 m L/min; and column temperature, 50 ℃. The gradient elution procedure was programmed as follows： 0–10 min, 95%–75% B; 10–15 min, 75% B; 15–20 min, 75%–65% B; 20–21 min, 65%–10% B; 21–24 min, 10% B, 24–25 min, 10%–95% B; and 25–35 min, 95% B. The results showed that the standard curves of 12 oligosaccharides had good linearity within the concentration range investigated（R2 0.98） and the recoveries were between 80.00% and 120.00%. The repeatability and instrumental precision（expressed as elative standard deviations） were less than 10%.