Separation of hexosamines, hexosaminitols and hexosamine-containing di- and trisaccharides on an amino acid analyser.
Separation of hexosamines, hexosaminitols and hexosamine-containing di- and trisaccharides on an amino acid analyser.
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在氨基酸分析仪上分离己糖胺、己糖胺醇以及含己糖胺的二糖和三糖。
DOI：
10.1016/s0021-9673(00)82587-x
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发表时间：
1977
期刊：
影响因子：
--
通讯作者：
A. S. Donald
中科院分区：
文献类型：
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作者：
A. S. Donald
二糖半乳糖L-+6 GlcNAc和乳寡糖由已故教授R.Kuhn(德国海德堡马克斯普朗克研究所)实验室获得。以N-内酯为原料，经部分酸解和纸层析分离得到GlcNAc#？(1+3)GAI。其他寡糖是在利斯特研究所(英国伦敦)的Winifred M.Watkins教授和WTJ Morgan教授的实验室中制备的。氨基半乳糖胺、氨基半乳糖胺、氨基半乳糖胺和氨基半乳糖醇如前所述获得。S_N-乙酰甘露糖胺购自Koch-ILight(英国科恩布鲁克)。以N-乙酰甘露糖胺酸解制备甘露糖胺溶液，N-乙酰甘露糖胺经NaBH还原酸解制备甘露糖醇溶液，低聚糖还原脱N-乙酰化反应如下：在5×0.4 cm管内的蔗糖溶液中加入10%(w/v)NaBH，3h后加入6M氢氧化钠(20~1)，在丙酮-CO中冷冻，真空封口。试管在100“的温度下加热16h，然后打开，在pH 2.2的缓冲液中装入1ml的内容物。将含有5-5Opg还原脱N-乙酰化糖的等分转移到氨基酸分析仪的自动配料中，用pH 2.2缓冲液洗涤，并按以下程序运行：缓冲液I(320分钟)，缓冲液II(200分钟)，0.5M氢氧化钠(40分钟)，缓冲液I(120分钟)。
The disaccharide Gal~(l-+ 6) GlcNAc and milk oligosaccharides were obtained from the laboratories of the late Professor R. Kuhn (Max-Planck Institute, Heideiburg, GFR). GlcNAc#?(1+ 3) GaI was prepared by partial acid hydrolysis of lacto-N-tetraose and isolation of the disaccharide by paper chromatography. Other oligosaccharides were prepared in the laboratories of Professors Winifred M. Watkins and WTJ Morgan at the Lister Institute (London, Great Britain). GIucosamine, galactosamine, gIucosamiuito1, and galactosaminitol were obtained as described previouslyS_ N-Acetylmannosamine was purchased from Koch-ILight (Colnbrook, Great Britain). Mannosamine solutions were prepared by acid hydrolysis of N-acetylmannosamine, and mannosaminitol solutions by NaBH., reduction and acid hydrolysis of N-acetylmannosamine.Oligosaccharides were reduced and de-N-acetylated as fohows: To the sugar solution (20 pg-1 mg in 90, uI water) in Pyrex tubes (5 x 0.4 cm ID) was added 10%(w/v) NaBH.,(10,~ l) _ After 3 h, 6 M NaOH (20~ 1) was added; the samples were frozen in acetone-solid CO, and then sealed under vacuum. The tubes were heated for 16 h at 100”, then opened, and the contents made up to 1 ml with pH 2.2 buffer. Aliquots containing 5--5Opg of the reduced de-N-acetylated sugars were transferred to the autofoader of the amino acid analyser, washed in with the pH 2.2 buffer, and run with the following programme: Buffer I (320 min), buffer II (200 min), 0.5 M NaOH (40 min), buffer I (120 min).