A method for the determination of N-acetylneuraminic acid (Neu5Ac) and N-glycolyneuraminic acid (Neu5Gc) in human milk and infant formulas was established by high-performance anion-exchange chromatography with pulsed amperometric detection. The sialic acids in human milk and infant formulas were released by the acid hydrolysis method, and the hydrolysate of infant formulas was purified by an ion chromatography pretreatment column. And Dionex CarboPac PA20-high performance anion chromatographic column was used for separation, gradient elution was conducted with sodium hydroxide and sodium acetate as eluents with the flow rate of 0.5 mL/min, injection volume of 10 μL, the column temperature of 30 ℃ and pulsed amperometric detector for detection. Furthermore, the acid hydrolysis time and the purification conditions of infant formulas were optimized, and the results showed that Neu5Ac and Neu5Gc could be separated well under the experimental conditions. And the ion peak area had a good linear relationship in a wide range of concentration (R2>0.999). The detection limits of Neu5Ac and Neu5Gc were 3 and 2 μg/L, and their limits of quantification were 10 and 6 μg/L, respectively. The spiked recoveries of sialic acids in human milk and infant formula were 87.65%-102.33%, the RSDs of stability were 1.86%-5.21%. This method developed here has the advantages of simple sample treatment procedure, low detection limit and quantitation limit, wide linear range, good stability, good repeatability and recovery rate. It can be widely used for the determination of sialic acids in milk substrate.
RU Yuanpu
,
CHEN Jun
,
ZHANG Minghui
,
QIAO Weicang
,
ZHAO Junying
,
HOU Juncai
,
CHEN Shuxing
,
CHEN Lijun
. Determination of sialic acids in human milk and infant formulas by high-performance anion-exchange chromatography with pulsed amperometric detection[J]. Food and Fermentation Industries, 2021
, 47(11)
: 221
-226
.
DOI: 10.13995/j.cnki.11-1802/ts.026527
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