Isolation and Structural Identification of Neutral Human Milk Oligosaccharides†

doi:10.7503/cjcu20150118

Abstract

Abstract:

Eighteen natural oligosaccharides were obtained from the defatted and deproteinated human milk using a combination isolation process including anion exchange chromatography(AEC), gel permeation chromatography(GPC) and porous graphitized carbon(PGC) liquid chromatography. Meanwhile, the structures of these oligosaccharides were identified by electrospray ionization collision-induced dissociation tandem mass spectrometry(ESI-CID-MS/MS). The major building blocks of neutral human milk oligosaccharides were consisted of tri- to nonasaccharides through the regular assembly of glucose(Glc), galactose(Gal), N-acetylglucosamine(GlcNAc) and fucose(Fuc). Lactose(Galβ1-4Glc) core was located at the reducing end in all of these oligosaccharides while the lactosamine(Galβ1-3/4GlcNAc) was determined at the non-reducing end. In addition, some of Gal, Glc and GlcNAc residues on the backbone of oligosaccharides were fucosylated in α1-2, α1-3 and α1-4 linkages, respectively.

Key words: Neutral human milk oligosaccharide, Isolation, ESI-CID-MS/MS

CLC Number:

O629.1

TrendMD:

LIU Shilong, LANG Yinzhi, ZHU He, YAN Lina, LÜ Youjing, ZHAO Xiaoliang, CAI Chao, YU Guangli. Isolation and Structural Identification of Neutral Human Milk Oligosaccharides^†[J]. Chem. J. Chinese Universities, 2015, 36(6): 1087.

Figures/Tables 6

Fig.1 Separation graph of neutral human milk oligosaccharides(NES) on Bio-Gel P2 column

Fig.2 Chromatograms of fractions na(A), nb(B) and nc(C) on HPLC-PGC column

Fig.3 Structures(A, B) and MS2 spectra(C, D) of deprotonated nb6(A, C) and nb7(B, D) at m/z 706- as precursor ion

Fig.4 Structures(A—C) and MS2 spectra(D—F) of deprotonated nb2(A, D), nb3(B, E) and nb5(C, F) at m/z 852- as precursor ionα: the large branch; β: the small branch.

Fig.5 Structures(A, B), MS2 and MS3 spectra(C—F) of nc2(A, C, E) and nc6(B, D, F)(C) MS2 spectrum of fraction nc2 at m/z 1509-; (D) MS2 spectrum of fraction nc6 at m/z 7542-; (E) MS3 spectrum of nc2 at m/z 836-; (F) MS3 spectrum of fraction nc6 at m/z 672-. α': the large branch of α; α″: the small branch of α; β': the large branch of β; β″: the small branch of β.

References 27

[1]	Wu S., Tao N., German J. B., Grimm R., Lebrilla C. B., J. Proteome Res., 2010, 9(8), 4138—4151
[2]	Bode L., Kuhn L., Kim H. Y., Hsiao L., Nissan C., Sinkala M., Kankasa C., Mwiya M., Thea D. M., Aldrovandi G. M., Am. J. Clin. Nutr., 2012, 96(4), 831—839
[3]	Kurakevich E., Hennet T., Hausmann M., Rogler G., Borsig L., Proc. Natl. Acad. Sci. USA, 2013, 110(43), 17444—17449
[4]	Jeurink P. V., Vanesch B. C., Rijnierse A., Garssen J., Knippels L. M ., Am. J. Clin. Nutr., 2013, 98(2), 572S—577S
[5]	Newburg D. S., J. Anim. Sci., 2009, 87(13), 26—34
[6]	Yu Z. T., Chen C., Newburg D. S., Glycobiology, 2013, 23(11), 1281—1292
[7]	Wang B., Miller B. J., Eur. J. Clin. Nutr., 2003, 57(11), 1351—1369
[8]	Hennet T., Weiss A., Borsig L., Swiss Med. Wkly., 2014, 144(w13927), 1—9
[9]	Totten S. M., Wu L. D., Parker E. A., Davis J. C., Hua S., Stroble C., Ruhaak L. R., Smilowitz J. T., German J. B., Lebrilla C. B., Anal. Bioanal. Chem., 2014, 406, 7925—7935
[10]	Galeotti F., Coppa G. V., Zampini L., Maccari F., Galeazz T., Padella. L., Santoro L., Gabrielli O., Volpi N., Electrophoresis, 2014, 35(6), 811—818
[11]	Totten S. M., Zivkovic A. M., Wu S., Ngyuen U. T., Freeman S. L., J. Proteome. Res., 2012, 11(12), 6124—6133
[12]	Blank D., Gebhardt S., Maass K., Lochnit G., Dotz V., Blank J., Geyer R., Kunz C., Anal. Bioanal. Chem., 2011, 401(8), 2495—2510
[13]	Deleoz M. L., Gaerlan S. C., Strum J. S., Dimapasoc L. M., Mirmiran M., Tancredi D. J., Smilowitz J. T., Kalanetra K. M., Mills D. A., German J. B., Lebrilla C. B., Underwood M. A., J. Proteome Res., 2012, 11(9), 4662—4672
[14]	Bao Y., Zhu L., Newburg D. S., Anal. Biochem., 2007, 370(2), 206—214
[15]	Kunz C., Rudloff S., Hintelmann A., Pohlentz G., Egge H., J. Chromatogr. B, 1996, 685, 211—221
[16]	Finke B., Stahl B., Pfenninger A., Karas M., Daniel H., Sawatzki G., Anal. Chem., 1999, 71(17), 3755—3762
[17]	Ruhaak L. R., Lebrilla C. B., Adv. Nutr., 2012, 3(3), 406S—414S
[18]	Strum J. B., Kim J., Wu S., Deleoz M. L., Egge H., Peacock. K., Grimm R., German J. B., Mills D., Lebrilla C. B., Anal. Chem., 2012, 84, 7793—7801
[19]	Galeotti F., Coppa G. V., Zampini L., Maccari F., Galeazz T., Padella L., Santoro L., Gabrielli O., Volpi N., Anal. Biochem., 2012, 430, 97—104
[20]	Dubois M., Gilles K. A., Hamilton J. K., Anal. Chem., 1956, 28(03), 350—356
[21]	Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J., J. Biol. Chem., 1951, 193(1), 265—275
[22]	Zhu H., Chen X., Zhang X., Liu L. L., Cong D. P., Zhao X., Yu G. L., Anal. Biochem., 2014, 456, 63—69
[23]	Wang P. P., Lü Y. J., Cao H., Zhao X. L., Li G. S., Wu J. D., Yu G. L., Chem. J. Chinese Universities, 2012, 33(8), 1722—1726
	(王培培, 吕友晶, 曹欢, 赵小亮, 李广生, 吴建东, 于广利.高等学校化学学报, 2012,33(8), 1722—1726 )
[24]	Chai W., Piskarev V., Lawson A. M., J. Am. Soc. Mass Spectrom., 2002, 13(6), 670—679
[25]	Chen H. H., Zhao X., Luan X. H., Yu G. L., Chem. J. Chinese Universities, 2015,36(1), 1—8 )
	(陈欢欢., 赵峡., 栾晓红., 于广利.高等学校化学学报, 2015,36(1), 1—8 )
[26]	Chai W., Piskarev V., Lawson A. M., Anal. Chem., 2001, 73(3), 651—657
[27]	West C., Elfakir C., Lafosse M., J. Chromatogr. A, 2010, 1217, 3201—3216