Determination of 2′-Fucosylactose and Lacto-N-Neotetraose Contents in Infant Formula by High Performance Anion Exchange Chromatography-Pulsed Amperometric Detector

doi:10.19894/j.issn.1000-0518.230216

Chinese Journal of Applied Chemistry ›› 2024, Vol. 41 ›› Issue (3): 405-414.DOI: 10.19894/j.issn.1000-0518.230216

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Determination of 2′-Fucosylactose and Lacto-N-Neotetraose Contents in Infant Formula by High Performance Anion Exchange Chromatography-Pulsed Amperometric Detector

Sheng-Qun ZHAN, Cheng GE(), Rong-Jie ZHOU, Jun ZHOU

Ausnutria Dairy （China） Co. Ltd. ，Changsha 410200，China

Received:2023-07-26 Accepted:2023-11-13 Published:2024-03-01 Online:2024-04-09
Contact: Cheng GE
About author:873685937@qq.com

Abstract

Abstract:

Simultaneous determination of 2'-fucosylactose and lacto-N-neotetraose in infant formula milk powder by ion chromatography is studied. These 2 human milk oligosaccharides （HMOs） are separated by ion chromatography， and determined by pulse amperometric detector， and quantified using an external standard method. The method is optimized through analyzing different purification methods， enzymatic hydrolysis treatment， different elution conditions， and the linear range. The effects of determining chromatographic conditions and pre-treatment conditions on separation are compared. The results show that the optimized method has a good linear relationship， with R²＞0.999. It also has good precision and accuracy， which the relative standard deviation （n=7） is ＜2.7%， and the recoveries of 2'-fucose lactose and lacto-N-neotetraose are 95.97%~103.38% and 100.09%~104.01%， respectively. This method can simultaneously determine 2'-fucose and lacto-N-neotetraose in infant formula milk powder. It is simple to operate， has good accuracy and precision， and can provide reference for the quality control method of infant formula milk powder.

Key words: Human milk oligosaccharides, Ion chromatography, 2'-Fucosylactose, Lactose-N-neotetraose, Infant milk powder

CLC Number:

O657.7

Sheng-Qun ZHAN, Cheng GE, Rong-Jie ZHOU, Jun ZHOU. Determination of 2′-Fucosylactose and Lacto-N-Neotetraose Contents in Infant Formula by High Performance Anion Exchange Chromatography-Pulsed Amperometric Detector[J]. Chinese Journal of Applied Chemistry, 2024, 41(3): 405-414.

Figures/Tables 10

References 19

1	ZIVKOVIC A M， GERMAN J B， LEBRILLA C B， et al. Human milk glycobiome and its impact on the infant gastrointestinal microbiota［J］. Proc Natl Acad Sci USA， 2011， 108（1）： 4653-4658.
2	WICIŃSKI M， SAWICKA E， GĘBALSKI J， et al. Human milk oligosaccharides： health benefits， potential applications in infant formulas， and pharmacology［J］. Nutrients， 2020， 12（1）： 266.
3	KOLETZKO B， BRANDS B， GROTE V， et al. Early nutrition programming project. long-term health impact of early nutrition： the power of programming［J］. Ann Nutr Metab， 2017， 70： 161-169.
4	BLACK R E， ALLEN L H， BHUTTA Z A， et al. Maternal and child undernutrition study group. maternal and child undernutrition： global and regional exposures and health consequences［J］. Lancet， 2008， 371： 243-260.
5	DIETERICH C M， FELICE J P， O′SULLIVAN E， et al. Breastfeeding and health outcomes for the mother-infant dyad［J］. Pediatr Clin N Am， 2013， 60： 31-48.
6	CHRISTENSEN A S， SKOV S H， LENDAL S E， et al. Quantifying the human milk oligosaccharides 2'-fucosyllactose and 3-fucosyllactose in different food applications by high performance liquid chromatography with refractive index detection［J］. J Food Sci， 2020， 85（2）： 332-339.
7	AUSTIN S， CUANY D， MICHAUD J， et al. Determination of 2'-fucosyllactose and lacto-n-neotetraose in infant formula［J］. Mol， 2018， 23（10）： 2650.
8	ELLINGSON D J， RUOSCH A J， FOSTER K L， et al. Analysis of six human milk oligosaccharides （HMO） in infant formula and adult nutritionals by 2AB labeling and quantification with HILIC-FLD： first action 2022.02［J］. J AOAC Int， 2022， 106（1）： 112-126.
9	AUSTIN S， DE CASTRO C A， BÉNET T， et al. Temporal change of the content of 10 oligosaccharides in the milk of Chinese urban mothers［J］. Nutrients， 2016， 8（6）： 346.
10	ZHANG W， WANG T， CHEN X， et al. Absolute quantification of twelve oligosaccharides in human milk using a targeted mass spectrometry-based approach［J］. Carbohydr Polym， 2019， 219： 328-333
11	FONG B， MA K， MCJARROW P. Quantification of bovine milk oligosaccharides using liquid chromatography-selected reaction monitoring-mass spectrometry［J］. J Agric Food Chem， 2011， 59（18）： 9788-9795.
12	BAO Y， CHEN C， NEWBURG D S. Quantification of neutral human milk oligosaccharides by graphitic carbon high-performance liquid chromatography with tandem mass spectrometry［J］. Anal Biochem， 2013， 433（1）： 28-35.
13	陈新新，芦晶，刘鹭，等. 超高效液相色谱-质谱法测定母乳中12种低聚糖［J］. 食品科学， 2018， 39（4）： 138-143.
	CHEN X X， LU J， LIU L， et al. Determination of 12 oligosaccharides in human milk by ultra-performance liquid chromatography-mass spectrometry［J］. Food Sci， 2018， 39（4）： 138-143.
14	MANK M， WELSCH P， HECK A J R， et al. Label-free targeted LC-ESI-MS2 analysis of human milk oligosaccharides （HMOs） and related human milk groups with enhanced structural selectivity［J］. Anal Bioanal Chem， 2019， 411（1）： 231-250.
15	NIJMAN R M， LIU Y， BUNYATRATCHATA A， et al. Characterization and quantification of oligosaccharides in human milk and infant formula［J］. J Agric Food Chem， 2018， 66（26）： 6851-6859.
16	陈磊， PHILIP H，田芳，等. 离子色谱法测定母乳中的寡聚糖与游离唾液酸［J］. 中国食品学报， 2019， 19（10）： 227-234.
	CHEN L， PHILIP H， TIAN F， et al. Determination of oligosaccharides and free sialic acid in human milk by ion chromatography［J］. J Chin Inst Food Sci Technol， 2019， 19（10）： 227-234.
17	GU F， KATE G A T， ARTS I C W， et al. Combining HPAEC-PAD， PGC-LC-MS， and 1D ¹H NMR to investigate metabolic fates of human milk oligosaccharides in 1-month-old infants： a pilot study［J］. J Agric Food Chem， 2021， 69（23）： 6495-6509.
18	AUER F， JARVAS G， GUTTMAN A. Recent advances in the analysis of human milk oligosaccharides by liquid phase separation methods［J］. J Chromatogr B， 2021， 1162： 122497.
19	尹大芳，孙晓杰，郭莹莹，等. 单糖定性定量的色谱分析方法的研究进展［J］. 食品工业科技， 2020， 41（24）： 321-329.
	YIN D F， SUN X J， GUO Y Y， et al. Development of qualitative and quantitative chromatographic analysis of monosaccharides［J］. Sci Technol Food Ind， 2020， 41（24）： 321-329.

Time/min	Mobile phase A/%	Mobile phase B/%	Mobile phase C/%	Mobile phase D/%
0.000	20	0	0	80
2.000	20	0	0	80
30.000	35	0	0	65
35.000	35	0	0	65
35.100	0	0	100	0
40.000	0	0	100	0
40.100	0	100	0	0
45.000	0	100	0	0
45.100	20	0	0	80
50.000	20	0	0	80

Time/min	Mobile phase A/%	Mobile phase B/%	Mobile phase C/%	Mobile phase D/%
0.000	20	0	0	80
2.000	20	0	0	80
30.000	35	0	0	65
35.000	35	0	0	65
35.100	0	0	100	0
40.000	0	0	100	0
40.100	0	100	0	0
45.000	0	100	0	0
45.100	20	0	0	80
50.000	20	0	0	80

		Apparatus precision（n=9）				Method precision（n=7）
		2'-FL		LNnT		2'-FL		LNnT
Items	Retention time/min	ρ/ （μg·mL^-1）	Peak area	Retention time/min	ρ/ （μg·mL^-1）	Peak area	Retention time/min	ρ/ （g·100 g^-1）	Retention time/min	ρ/ （g·100 g^-1）
Mean	17.867 1	3.893 3	12.198 9	29.444 8	1.973 5	3.802 9	17.758	1.145 5	29.473 0	0.252 2
SD	0.045 0	0.072 5	0.227 6	0.080 5	0.038 1	0.076 7	0.032	0.021 0	0.048	0.005 1
CV/%	0.25	1.86	1.87	0.27	1.93	2.02	0.18	1.83	0.16	2.03

		Apparatus precision（n=9）				Method precision（n=7）
		2'-FL		LNnT		2'-FL		LNnT
Items	Retention time/min	ρ/ （μg·mL^-1）	Peak area	Retention time/min	ρ/ （μg·mL^-1）	Peak area	Retention time/min	ρ/ （g·100 g^-1）	Retention time/min	ρ/ （g·100 g^-1）
Mean	17.867 1	3.893 3	12.198 9	29.444 8	1.973 5	3.802 9	17.758	1.145 5	29.473 0	0.252 2
SD	0.045 0	0.072 5	0.227 6	0.080 5	0.038 1	0.076 7	0.032	0.021 0	0.048	0.005 1
CV/%	0.25	1.86	1.87	0.27	1.93	2.02	0.18	1.83	0.16	2.03

Mass/g	2'-FL			LNnT
Mass/g	Adding mass/mg	ρ/（μg·mL^-1）	Recovery/%	Adding mass/mg	ρ/（μg·mL^-1）	Recovery/%
2.019 5	23.990 4	7.912 3	98.94	12.047 3	4.068 6	101.32
2.042 1		8.227 0	102.88		4.118 8	102.57
2.005 5		8.267 4	103.38		4.147 3	103.28
2.003 7	15.993 6	5.313 3	99.66	8.031 5	2.734 4	102.14
2.045 9		5.409 7	101.47		2.784 4	104.01
2.000 5		5.296 4	99.35		2.723 1	101.72
2.040 5	7.996 8	2.558 3	95.97	4.015 8	1.347 2	100.64
2.009 1		2.633 4	98.79		1.376	102.79
2.008 5		2.558 7	95.99		1.339 8	100.09

Determination of 2′-Fucosylactose and Lacto-N-Neotetraose Contents in Infant Formula by High Performance Anion Exchange Chromatography-Pulsed Amperometric Detector

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