唾液酸化人乳寡糖的分离及串联质谱结构序列分析

doi:10.7503/cjcu20170593

高等学校化学学报 ›› 2018, Vol. 39 ›› Issue (4): 645.doi: 10.7503/cjcu20170593

唾液酸化人乳寡糖的分离及串联质谱结构序列分析

郎银芝, 刘世龙, 王晨, 张晓, 吕友晶, 蔡超, 李国云, 于广利()

山东省糖科学与糖工程重点实验室, 海洋药物教育部重点实验室, 中国海洋大学医药学院, 青岛 266003

收稿日期:2017-08-31 出版日期:2018-04-10 发布日期:2018-03-18
作者简介:联系人简介: 于广利, 男, 博士, 教授, 博士生导师, 主要从事糖化学及糖药物学方面的研究. E-mail:glyu@ouc.edu.cn
基金资助:
国家自然科学基金(批准号: 31670811)、国家自然科学基金山东海洋科学研究中心联合基金(批准号: U1406403)、山东省科技项目(批准号: 2015ZDJS04002, LKH2015102)和泰山学者工程专项基金资助

Separation and Structural Sequence Analysis of Sialylated HMOs via Tandem Mass Spectrometry^†

LANG Yinzhi, LIU Shilong, WANG Chen, ZHANG Xiao, LÜ Youjing, CAI Chao, LI Guoyun, YU Guangli^*()

Shandong Provincial Key Laboratory of Glycoscience and Glycotechnology, Key Laboratory of Marine Drugs,Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China

Received:2017-08-31 Online:2018-04-10 Published:2018-03-18
Contact: YU Guangli E-mail:glyu@ouc.edu.cn
Supported by:
† Supported by the National Natural Science Foundation of China(No.31670811), the National Natural Science Foundation of China-Shandong Joint Fund for Marine Science Research Centers(No.U1406403), the Science & Technology Projects of Shandong Province, China(Nos.2015ZDJS04002, LKH2015102) and the Taishan Scholar Project Special Funds of China

摘要/Abstract

摘要：

基于不同唾液酸化寡糖之间的极性、空间结构及电荷密度的差异性, 采用半制备型多孔性石墨化碳(PGC)色谱分离制备了高纯度唾液酸化人乳寡糖单体, 并使用离线的电喷雾-串联质谱技术(ESI-MSⁿ)确定8种寡糖单体的序列结构分别为6'-SL, 3'-SL, LSTb, LSTc, F-LSTc, F-LSTb, DSLNT与FS-LNnH. 本文结果为进一步从事唾液酸化人乳寡糖的结构与活性研究提供了参考依据.

关键词: 唾液酸化人乳寡糖, 石墨化碳色谱分离, 串联质谱, 结构序列

Abstract:

Sialylated human milk oligosaccharides(SHMOs), important components of human milk oligosaccharides, play a key role in the growth and development of the newborn. Herein, based on the difference of polarity, spatial structure and charge density of sialylated oligosaccharides, a semi-preparative porous graphitized carbon(PGC) chromatography was employed to separate SHMOs in high purity. An off-line electrospray ionization tandem mass spectrometry(ESI-MSⁿ) was performed to identify the sequential structures of eight oligosaccharides including 6'-SL, 3'-SL, LSTb, LSTc, F-LSTc, F-LSTb, DSLNT and FS-LNnH. This study provides a useful reference for the further structure-activity analysis and application development of SHMOs.

Key words: Sialylated human milk oligosaccharides, Porous graphitized carbon separation, Tandem MS, Sequence analysis

中图分类号:

O657
O629.1

TrendMD:

郎银芝, 刘世龙, 王晨, 张晓, 吕友晶, 蔡超, 李国云, 于广利. 唾液酸化人乳寡糖的分离及串联质谱结构序列分析. 高等学校化学学报, 2018, 39(4): 645.

LANG Yinzhi, LIU Shilong, WANG Chen, ZHANG Xiao, LÜ Youjing, CAI Chao, LI Guoyun, YU Guangli. Separation and Structural Sequence Analysis of Sialylated HMOs via Tandem Mass Spectrometry^†. Chem. J. Chinese Universities, 2018, 39(4): 645.

图/表 8

Fig.1 Determination of sialic acids in NHMOs and SHMOs via an improved RP-HPLC methoda. Standard; b. SHMOs; c. NHMOs.

Fig.2 Separation of eight sialylated human milk oligosaccharides by PGC-HPLC method(A) and their ESI-MS spectra(B)

Table 1 Structural determination of eight sialylated human milk oligosaccharides by ESI-MSn

Fraction	Molecular ion	Theoretical M_w	DP^*	Composition	Glycan name
S1	632.19[M-H]^-	633.21	3	Neu5Ac₁Gal₁Glc₁	6'-SL
S2	632.19[M-H]^-	633.21	3	Neu5Ac₁Gal₁Glc₁	3'-SL
S4	997.31[M-H]^-	998.34	5	Neu5Ac₁Gal₂Glc₂	LSTb
S6	997.31[M-H]^-	998.34	5	Neu5Ac₁Gal₂Glc₂	LSTc
S5	1143.36[M-H]^-	1144.40	6	Fuc₁Neu5Ac₁Gal₂Glc₂	F-LSTc
S7	1143.36[M-H]^-	1144.40	6	Fuc₁Neu5Ac₁Gal₂Glc₂	F-LSTb
S3	643.70[M-2H]^2-	1289.44	6	Neu5Ac₂Gal₂Glc₂	DSLNT
S8	1508.48[M-H]^-; 771.74[M+2H₂O-2H]^2-	1509.53	8	Fuc₁ Neu5Ac₁Gal₃Glc₃	FS-LNnH

Fig.3 ESI-MS/MS spectra of monosialylated lactose species from S1(A) and S2(B) using [M-H]-(m/z 632.19) as the precursor ions

Fig.4 ESI-MS/MS spectra of monosialylated tetrasaccharide species from S4(A) and S6(B) using [M-H]-(m/z 997.31) as the precursorsα: The large branch; β: the small branch.

Fig.5 ESI-MS/MS spectra of monofucosylated and monosialylated tetrasaccharide species from S7(A) and S5(B) using [M-H]-(m/z 1143.36) as the precursor ionsα: The large branch; β: the small branch.

Fig.6 ESI-MS/MS product ion spectra and ESI-MS3 product ion spectrum of disialylated tetrasaccharide from S3 fraction using [M-H]-(m/z 1288.40)(A) and Y1α(m/z 997.31)(B) as the precursor,respectivelyα: The large branch; β: the small branch.

Fig.7 ESI-MS/MS and ESI-MS3 spectrum of monofucosylated and monosialylated hexasaccharide from S8 fraction using[M-H]-(m/z 1508.48) and D2b-4(m/z 673.21) as the precursor, respectivelyα, The large branch. β, the small branch; β', the large branch of β; β'', the small branch of β.

参考文献 21

[1]	Bode L., Glycobiology, 2012, 22, 1147-1162
[2]	Mantovani V., Galeotti F., Maccari F., Volpi N., Electrophoresis,2016, 37, 1514-1524
[3]	Oliveira D. L., Wilbey R. A., Grandison A. S., Roseiro L. B., Int. J. Dairy Technol., 2015, 68, 305-321
[4]	Urashima T., Kitaoka M., Asakuma S., Messer M., Comprehensive Glycoscience, 2007, 42, 695-724
[5]	Bode L., Jantscher-Krenn E., Adv. Nutr., 2012, 3, 383S-391S
[6]	Varki A., Trends Mol. Med., 2008, 14, 351-360
[7]	Varki A., Gagneux P., Ann. N. Y. Acad. Sci., 2012, 1253, 16-36
[8]	Wang B., Yu B., Karim M., Hu H., Sun Y., Mcgreevy P., Petocz P., Held S., Miller J. B., Am. J. Clin. Nutr., 2007, 85, 561-569
[9]	Liu S. L., Lang Y. Z., Zhu H., Yan L. N., Lü Y. J., Zhao X. L., Cai C., Yu G. L., Chem. J. Chinese Universities, 2015, 36(6), 1087-1093
	(刘世龙, 郎银芝, 祝贺, 闫立娜, 吕友晶, 赵小亮, 蔡超, 于广利. 高等学校化学学报, 2015, 36(6), 1087-1093)
[10]	Ito M., Ikeda K., Suzuki Y., Tanaka K., Saito M., Anal. Biochem., 2002, 300, 260-266
[11]	Spichtig V., Michaud J., Austin S., Anal. Biochem., 2010, 405, 28-40
[12]	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)
[13]	Lang Y., Zhao X., Liu L., Yu G., Mar. Drugs, 2014, 12, 4005-4030
[14]	Spengler B., Dolce J. W., Cotter R. J., Anal. Chem., 1990, 62, 1731-1737
[15]	Hofmeister G. E., Zhou Z., Leary J. A., J. Am. Chem. Soc., 1991, 113, 5964-5970
[16]	Wu S., Grimm R., German J. B., Lebrilla C. B., J. Proteome Res., 2011, 10, 856-868
[17]	Wu S., Tao N., German J. B., Grimm R., Lebrilla C. B., J. Proteome Res., 2010, 9, 4138-4151
[18]	Donmon B., Costello C. E., Glycoconj. J., 1988, 5, 397-409
[19]	Chai W., Piskarev V., Lawson A. M., J. Am. Soc. Mass Spectrom., 2002, 13, 670-679
[20]	Chai W., Piskarev V., Lawson A. M., Anal. Chem., 2001, 73, 651-657
[21]	Wheeler S. F., Harvey D. J., Anal. Chem., 2000, 72, 5027-5039

唾液酸化人乳寡糖的分离及串联质谱结构序列分析

Separation and Structural Sequence Analysis of Sialylated HMOs via Tandem Mass Spectrometry^†

RichHTML

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 21

相关文章 15

编辑推荐

Metrics

本文评价

[1]	杜晨晖, 李泽, 崔小芳, 张敏, 裴香萍, 詹海仙, 闫艳. 基于UPLC-Q-Orbitrap MS/MS技术研究酸枣仁发酵过程中的化学成分转化[J]. 高等学校化学学报, 2019, 40(8): 1614.
[2]	刘世龙, 郎银芝, 祝贺, 闫立娜, 吕友晶, 赵小亮, 蔡超, 于广利. 中性人乳寡糖的分离及结构鉴定[J]. 高等学校化学学报, 2015, 36(6): 1087.
[3]	皮子凤, 王倩倩, 张静, 宋凤瑞, 刘志强. 北五味子对糖尿病脑病大鼠脑中神经活性物质含量影响的在线微透析-高效液相色谱-串联质谱联用分析[J]. 高等学校化学学报, 2015, 36(3): 442.
[4]	陈海燕, 丁兰, 刘密兰. 微波辅助合成分子印迹聚合物用于萃取蜂蜜中的氯霉素[J]. 高等学校化学学报, 2015, 36(1): 67.
[5]	张洪涛, 朱莉, 詹晓北. 基于电喷雾串联质谱的蓝藻寡糖脱果糖分析及系列α-1,2-葡聚寡糖的制备[J]. 高等学校化学学报, 2014, 35(12): 2547.
[6]	张洪涛, 朱莉, 张爽, 詹晓北. 基于ESI-CID-MS/MS离子碎片峰特征的香菇多糖水解机制及系列香菇寡糖的制备[J]. 高等学校化学学报, 2014, 35(11): 2329.
[7]	毕云枫, 刘舒, 李雪, 刘志强, 宋凤瑞. 乌头类生物碱组分在CYP450中的代谢指纹图谱及对CYP450活性的影响[J]. 高等学校化学学报, 2013, 34(9): 2084.
[8]	李雪, 方小伟, 李银萍, 陈焕文. 羟基多环芳烃的电喷雾电离离子阱串联质谱检测[J]. 高等学校化学学报, 2013, 34(8): 1840.
[9]	黄宝勇, 欧阳喜辉, 孙江, 肖志勇, 潘灿平. 大气压固体分析探头离子源-串联质谱法快速检测蔬菜中多种农药残留[J]. 高等学校化学学报, 2013, 34(7): 1591.
[10]	潘素素, 赵娜, 欧阳永中, 黄科科, 丁健桦, 陈焕文, 袁龙, 王兴祥. 土壤中一种新的尿素衍生物的电喷雾萃取电离质谱分析[J]. 高等学校化学学报, 2013, 34(6): 1379.
[11]	毕云枫, 朱洪彬, 皮子凤, 刘志强, 宋凤瑞. UPLC-MS/MS结合多探针底物方法研究刺五加叶中黄酮苷类成分对CYP450活性的影响[J]. 高等学校化学学报, 2013, 34(5): 1067.
[12]	曹慧, 陈小珍, 朱岩, 李祖光. 多壁碳纳米管固相萃取技术同时测定蜂蜜中多类兽药残留[J]. 高等学校化学学报, 2013, 34(12): 2710.
[13]	王培培, 吕友晶, 曹欢, 赵小亮, 李广生, 吴建东, 于广利. 杂合褐藻糖胶寡糖的制备及结构分析[J]. 高等学校化学学报, 2012, 33(08): 1722.
[14]	丁健桦王兴祥张慧潘素素罗明标李建强陈焕文. 芹菜素的电喷雾萃取电离串联质谱[J]. 高等学校化学学报, 2011, 32(8): 1714.
[15]	符继红李俊芳付佩岳凡张旭龙王吉德. 组氨酸钴氧合反应的电喷雾串联质谱[J]. 高等学校化学学报, 2011, 32(7): 1483.

唾液酸化人乳寡糖的分离及串联质谱结构序列分析

Separation and Structural Sequence Analysis of Sialylated HMOs via Tandem Mass Spectrometry†

RichHTML

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 21

相关文章 15

编辑推荐

Metrics

本文评价

Separation and Structural Sequence Analysis of Sialylated HMOs via Tandem Mass Spectrometry^†