Giuffrida Francesca, Elmelegy Isabelle Masserey, Thakkar Sagar K, Marmet Cynthia, Destaillats Frédéric
Nestlé Research Center, Nestec Ltd, P.O. Box 44, Vers-chez-les-Blanc, 26, 1000, Lausanne, Switzerland,
Lipids. 2014 Oct;49(10):997-1004. doi: 10.1007/s11745-014-3943-2. Epub 2014 Sep 4.
It has been reported that dietary gangliosides may have an important role in preventing infections and in brain development during early infancy. However, data related to the evolution of their concentration over the different stages of lactation are scarce. Liquid chromatography coupled with electrospray ionization high resolution mass spectrometer (LC/ESI-HR-MS) has been optimized to quantify the two major ganglioside classes, i.e., aNeu5Ac(2-8)aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer (GD3) and aNeu5Ac(2-3)bDGalp(1-4)bDGlcp(1-1)Cer (GM3) in human milk. Gangliosides were extracted using chloroform and methanol, further purified by solid-phase extraction and separated by reversed-phase liquid chromatography. Repeatability, intermediate reproducibility, and recovery values were assessed to validate the method. In human milk, GD3 and GM3 could be quantified at the level of 0.1 and 0.2 μg/mL, respectively, with relative standard deviation of repeatability [CV(r)] and intermediate reproducibility [CV(iR)] values ranging from 1.9 to 15.0 % and 1.9 to 22.5 %, respectively. The described method was used to quantify GD3 and GM3 in human milk samples collected from 450 volunteers between 0 and 11 days and at 30, 60 and 120 days postpartum, providing for the first time the concentration of these minor lipids in a large cohort. The content of total gangliosides ranged from 8.1 and 10.7 μg/mL and the mean intake of gangliosides in infants 30, 60 and 120 days postpartum could be estimated at about 5.5, 7.0 and 8.6 mg of total gangliosides per day, respectively, when infants were exclusively breastfed.
据报道，膳食神经节苷脂可能在预防感染以及婴儿早期大脑发育中发挥重要作用。然而，关于其在哺乳期不同阶段浓度变化的数据却很稀少。液相色谱联用电喷雾电离高分辨率质谱仪（LC/ESI-HR-MS）已得到优化，用于定量母乳中两种主要的神经节苷脂类别，即α-Neu5Ac(2-8)α-Neu5Ac(2-3)β-DGalp(1-4)β-DGlcp(1-1)Cer（GD3）和α-Neu5Ac(2-3)β-DGalp(1-4)β-DGlcp(1-1)Cer（GM3）。神经节苷脂采用氯仿和甲醇进行提取，通过固相萃取进一步纯化，并通过反相液相色谱进行分离。对方法的重复性、中间精密度和回收率进行了评估以验证该方法。在母乳中，GD3和GM3的定量限分别为0.1和0.2 μg/mL，重复性[CV(r)]和中间精密度[CV(iR)]的相对标准偏差值分别在1.9%至15.0%和1.9%至22.5%之间。所描述的方法用于对450名志愿者在产后0至11天以及30、60和120天收集的母乳样本中的GD3和GM3进行定量，首次提供了一个大样本队列中这些微量脂质的浓度。总神经节苷脂含量在8.1至10.7 μg/mL之间，当婴儿纯母乳喂养时，产后30、60和120天婴儿每天神经节苷脂的平均摄入量分别约为5.5、7.0和8.6 mg总神经节苷脂。
