Determination of disialoganglioside GD3 and monosialoganglioside GM3 in infant formulas and whey protein concentrates by ultra‐performance liquid chromatography/electrospray ionization tandem mass spectrometry

Abstract

A method of ultra-performance liquid chromatography combined with electrospray ionization triple quadrupole tandem mass spectrometry (UPLC-ESI-MS/MS) has been established for simultaneous determination of major disialoganglioside 3 (GD3) and monosialoganglioside 3 (GM3) in infant formulas and whey protein concentrates. Gangliosides were extracted by using the technique of Svennerholm and Fredman and then cleaned up with OASIS HLB solid-phase extraction (SPE) cartridges. The various molecular species of gangliosides were separated on an Acquity UPLC BEH C8 column and analyzed under the negative ion mode. GD3 and GM3 were rapidly quantified using internal standard (IS) method. The developed method was further validated by determining the linearity, average recovery, sensitivity (limit of quantification), and precision. The results presented high correlation coefficients (R(2) > 0.993) of the selected 16 gangliosides molecular species and provided the respective linear ranges. The limit of quantification was 0.325-0.734 mg/100 g for eight molecular species of GD3 and 0.008-0.312 mg/100 g for eight molecular species of GM3, respectively. The reasonable average recoveries (81-95%) and precision (relative standard deviation [RSD] ≤15%) were also demonstrated in three different spiked levels. This new method would be very useful in the quantitative determination of gangliosides in infant formulas and whey protein concentrates.

The Role of Gangliosides in Neurodevelopment

“…The first evidence in humans that neonatal brain ganglioside concentration could be influenced by diet was provided in a study of infants who died of sudden infant death syndrome, in which it was found that those babies who had been largely breastfed had a 32% higher ganglioside content (measured as ganglioside-bound sialic acid) in their frontal cortices than formula fed babies [ 111 ]. Human milk naturally contains gangliosides, and although cow’s milk, which provides the basis for most infant formula (IF), contains similar amounts of ganglioside to human breast milk, ganglioside concentration can be substantially lower in unsupplemented IF and indeed, gangliosides may be completely absent from soy-based IF [ 30 , 112 , 113 , 114 ]. This implies that ganglioside intake is generally lower in IF-fed infants than in breast-fed infants.…”

Profiling gangliosides from milk products and other biological membranes using LC/MS

“…The third quadrupole is set to allow sialic acid to pass and to collect fragments. Triple quadrupole MS with MRM has been recently used due to the high specificity for simultaneous detection of multiple components in complex samples (Zhang et al, 2012;Lee et al, 2013;Giuffrida et al, 2014;Huang et al, 2014).…”

Mass‐spectrometry‐based lipidomics

“…extracted the total lipid fraction from adipose tissue using the modified Bligh–Dyer method and further removed neutral lipids from the total lipid extract with a silica SPE cartridge to yield FAHFAs . C8 and C18 cartridges have been used to isolate cerebrosides, gangliosides, eicosanoids, steroid hormones and FAs from polar matrixes in biofluid samples . 96‐well SPE plates enable pretreatment of 96 samples at once, which enables improved automation, throughput and parallel mode of lipid extractions .…”