A sensitive and efficient method for determination of N-acetylhexosamines and N-acetylneuraminic acid in breast milk and milk-based products by high-performance liquid chromatography via UV detection and mass spectrometry identification

doi:10.1016/j.jchromb.2015.12.032

NWIPB OpenIR

	A sensitive and efficient method for determination of N-acetylhexosamines and N-acetylneuraminic acid in breast milk and milk-based products by high-performance liquid chromatography via UV detection and mass spectrometry identification
	Wu, Chuanxiang 1; Xia, Lian 1; Liu, Lijie 1; Qu, Fengli 1; Sun, Zhiwei 1; Zhao, Xianen 1; You, Jinmao 1,2
	2016-02-01
发表期刊	JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
卷号	1011 页码:14-23
文章类型	Article
摘要	A sensitive and efficient method of high performance liquid chromatography using 1-(2-naphthyl)-3-methyl-5-pyrazolone (NMP) as pre-column derivatization reagent coupled with UV detection (HPLC-UV) and online mass spectrometry identification was established for determination of the most common N-Acetylhexosamines (N-acetyl-D-glucosamine (GlcNAc) and N-acetyl-n-galactosamine (GalNAc)) and N-acetylneuraminic acid (Neu5Ac). In order to obtain the highest liberation level of the three monosaccharides without destruction of Neu5Ac or conversion of GlcNAc/GaINAc to GlcN/GalN in the hydrolysis procedure, the pivotal parameters affecting the liberation of N-acetylhexosamines/Neu5Ac from sample were investigated with response surface methodology (RSM). Under the optimized condition, maximum yield was obtained. The effects of key parameters on derivatization, separation and detection were also investigated. At optimized conditions, three monosaccharides were labeled fast and entirely, and all derivatives exhibited a good baseline resolution and high detection sensitivity. The developed method was linear over the calibration range 0.25-12 mu M, with R-2 > 0.9991. The detection limits of the method were between 0.48 and 2.01 pmol. Intra- and inter-day precisions for the three monosaccharides (GlcNAc, GalNAc and Neu5Ac) were found to be in the range of 3.07-4.02% and 3.69-4.67%, respectively. Individual monosaccharide recovery from spiked milk was in the range of 81%-97%. The sensitivity of the method, the facility of the derivatization procedure and the reliability of the hydrolysis conditions suggest the proposed method has a high potential for utilization in routine trace N-acetylhexosamines and Neu5Ac analysis in biological samples. (C) 2015 Elsevier B.V. All rights reserved.
关键词	N-acetylhexosamines N-acetylneuraminic Acid Pre-column Derivatization 1-(2-naphthyl)-3-methyl-5-pyrazolone Hplc-uv-ms/ms Milk-based Products
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
DOI	10.1016/j.jchromb.2015.12.032
关键词[WOS]	C-TYPE LECTIN ; ACETYL-D-GLUCOSAMINE ; MONOSACCHARIDE COMPOSITION ; DERIVATIZATION ; HPLC ; ACETYLGALACTOSAMINE ; OLIGOSACCHARIDES ; POLYSACCHARIDES ; QUANTIFICATION ; OSTEOARTHRITIS
收录类别	SCI
语种	英语
项目资助者	National Natural Science Foundation of China(21505084 ; Natural Science Foundation of Shandong Province(ZR2014BM029 ; 21275089 ; ZR2012BQ022) ; 21475075 ; 21375076 ; 21305076 ; 81303179)
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
WOS类目	Biochemical Research Methods ; Chemistry, Analytical
WOS记录号	WOS:000369458600003
引用统计	被引频次：8[WOS] [WOS记录] [WOS相关记录]
文献类型	期刊论文
条目标识符	http://210.75.249.4/handle/363003/6494
专题	中国科学院西北高原生物研究所
作者单位	1.Qufu Normal Univ, Key Lab Life Organ Anal Shandong Prov, Qufu 273165, Peoples R China 2.Chinese Acad Sci, Northwest Plateau Inst Biol, Xining 810001, Peoples R China
推荐引用方式 GB/T 7714	Wu, Chuanxiang,Xia, Lian,Liu, Lijie,et al. A sensitive and efficient method for determination of N-acetylhexosamines and N-acetylneuraminic acid in breast milk and milk-based products by high-performance liquid chromatography via UV detection and mass spectrometry identification[J]. JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,2016,1011:14-23.
APA	Wu, Chuanxiang.,Xia, Lian.,Liu, Lijie.,Qu, Fengli.,Sun, Zhiwei.,...&You, Jinmao.(2016).A sensitive and efficient method for determination of N-acetylhexosamines and N-acetylneuraminic acid in breast milk and milk-based products by high-performance liquid chromatography via UV detection and mass spectrometry identification.JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES,1011,14-23.
MLA	Wu, Chuanxiang,et al."A sensitive and efficient method for determination of N-acetylhexosamines and N-acetylneuraminic acid in breast milk and milk-based products by high-performance liquid chromatography via UV detection and mass spectrometry identification".JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES 1011(2016):14-23.