利用UPLC-TOF-MS测定成熟母乳及婴幼儿配方奶粉中主要神经节苷酯含量

Determination of content of main gangliosides in mature human milk and infant formula by UPLC-TOF-MS

建立了超高效液相色谱结合飞行时间质谱法分析成熟母乳中主要神经节苷酯双唾液酸神经节苷酯GD3和单唾液酸神经节苷酯GM3的含量。通过C18柱将不同结构的神经节苷酯进行分离,并在ESI负离子模式下进行分析。用外标法对GD3和GM3进行了定量,并对该方法进行方法学验证。结果表明:神经节苷酯GD3、GM3分别在5~200μg/mL和0.1~10.0μg/mL范围内线性关系良好,相关系数均大于等于0.9620,神经节苷酯GD3和GM3的定量限分别为2.75μg/mL和0.29μg/mL,加标回收率为92.15%~111.78%,精密度小于等于6.07%。成熟母乳中神经节苷酯GD3和GM3含量分别为(9.61±3.32)μg/mL和(2.08±1.39)μg/mL;4种婴幼儿配方奶粉中GD3含量略高于成熟母乳,但未检出GM3。

An ultra-performance liquid chromatography combined with time-of-flight mass spectrometry method was established to analyze the contents of the main gangliosides(disialoganglioside GD3 and monosialoganglioside GM3)in mature human milk.The gangliosides of different molecular species were separated by C18 column and analyzed in ESI negative ion mode.The external standard method was used to quantify GD3 and GM3,and the method was validated.The results showed that gangliosides GD3 and GM3 had a good correlation coefficient(R^(2)≥0.9620)in the range of 5-200μg/mL and 0.1-10.0μg/mL,respectively.The limits of quantification of gangliosides GD3 and GM3 were 2.75μg/mL and 0.29μg/mL,respectively.They also demonstrated reasonable recovery rates(92.15%-111.78%)and precision(relative standard deviation≤6.07%).The contents of gangliosides GD3 and GM3 in mature human milk were(9.61±3.32)μg/mL and(2.08±1.39)μg/mL,respectively.The content of gangliosides GD3 in 4 infant formulas was slightly higher than that in mature human milk,but gangliosides GM3 was not detected in all infant formulas.