Primary Structural Analysis of Dendrobium officinale Polysaccharide†

doi:10.7503/cjcu20170464

Abstract

Abstract:

A homogeneous polysaccharide(SDOP) was obtained from the fresh Dendrobium officinale. The relative molecular mass and composition of monosaccharide were then determined by gel permeation chromatography-laser scatter(GPC-LS) and high performance liquid chromatography with pre-column derivatization using 1-phenyl-3-methyl-5-pyrazolone(PMP) as derivatization agent. The detailed chemical structure of SDOP was elucidated by means of infrared spectroscopy, methylation analysis and nuclear magnetic resonance spectroscopy(NMR). The result shows that SDOP with an M_w of 1.66×10⁶ consists dominantly of mannose and glucose in a molar ratio of 4.9∶1.0. The results of NMR and methylation analysis suggest that SDOP is a 1,4-linked O-acetylated glucomannan, with a small amount of branch and end signal, while acetyl groups are mainly attached to the O-2 or O-3 positions on saccharide ring.

Key words: Dendrobium officinale, Polysaccharide, Structure analysis, Nuclear magnetic resonance

CLC Number:

O629

TrendMD:

GAO Yunxiao,HU Xiaolong,WANG Yuerong,JIANG Zhihong,ZHANG Hongyang,ZHANG Min,HU Ping. Primary Structural Analysis of Dendrobium officinale Polysaccharide^†[J]. Chem. J. Chinese Universities, 2018, 39(5): 934.

Figures/Tables 4

Table 1 Methylation analysis result of SDOP

t_R/min	Name of partially methylate alditol acetate	Linkage
15.155	1,5-Di-O-acetyl-2,3,4,6-tetra-O-methylhexitol	Manp-(1→
15.240	1,5-Di-O-acetyl-2,3,4,6-tetra-O-methylhexitol	Glcp-(1→
17.200	1,4,5-Tri-O-acetyl-2,3,6-tri-O-methylhexitol	→4)-Manp-(1→
17.305	1,4,5-Tri-Oacetyl-2,3,6-tri-O-methylhexitol	→4)-Glcp-(1→
18.415	1,3,4,5-Tetra-O-acetyl-2,6-di-methylhexitol	→3,4)-Manp-(1→
18.490	1,3,4,5-Tetra-O-acetyl-2,6-di-methylhexitol	→3,4)-Glcp-(1→
18.865	1,2,4,5-Tetra-O-acetyl-3,6-di-O-methylhexitol	→2,4)-Manp-(1→
19.305	1,4,5,6-Tetra-O-acetyl-2,3-di-O-methylhexitol	→4,6)-Manp-(1→
19.380	1,4,5,6-Tetra-O-acetyl-2,3-di-O-methylhexitol	→4,6)-Glcp-(1→
19.540	1,2,3,4,5-Penta-O-acetyl-6-O-methylhexitol	→2,3,4)-Manp-(1→
20.485	1,2,3,4,5-Penta-O-acetyl-6-O-methylhexitol	→2,3,4)-Glcp-(1→
20.580	1,3,4,5,6-Penta-O-acetyl-2-O-methylhexitol	→3,4,6)-Manp-(1→
20.885	1,2,4,5,6-Penta-O-acetyl-3-O-methylhexitol	→2,4,6)-Manp-(1→
21.025	1,2,3,5,6-Penta-O-acetyl-4-O-methylhexitol	→2,3,6)-Manp-(1→

Fig.1 HSQC spectrum(A), COSY spectrum(B), TOCSY spectrum(C) and HMBC spectrum(D) of enzymatic hydrolysis product of SDOP

Table 2 1H and 13C chemical shifts(δ) for SDOP

Residue	M	M₂	M₃	M₂₃	G	M_α	M_β
1	4.77,102.94	4.96,101.95	4.84,102.39	5.05,101.33	4.52,105.33	5.19,96.71	4.92,96.60
2	4.13,72.81	5.52,74.50	4.20,71.54	5.53,72.38	3.38,76.72	4.01,71.74	4.00,73.49
3	3.82,74.28	4.00,73.09	5.11,76.24	5.19,74.63	3.72,76.78	3.91	3.91
4	3.79,79.30	3.84,79.36	4.05,75.97	4.10,74.65	3.70,81.30	4.05
5	3.57,77.84	3.59,77.91	3.65,75.65	3.72,72.47	3.66,72.49	3.62	3.54
6	3.92,63.31	3.89,63.18	3.90,63.34	3.94	3.91,63.12	3.94	3.88
6'	3.76	3.79	3.75	3.78
Linkage	G/M/M₂/M₃/ $M 2 3$ /M_α	G/M	G/M	M/M₂/M₃	G/M/M₂/M₃/M_β

Table 2 1H and 13C chemical shifts(δ) for SDOP

Residue	M	M₂	M₃	M₂₃	G	M_α	M_β
1	4.77,102.94	4.96,101.95	4.84,102.39	5.05,101.33	4.52,105.33	5.19,96.71	4.92,96.60
2	4.13,72.81	5.52,74.50	4.20,71.54	5.53,72.38	3.38,76.72	4.01,71.74	4.00,73.49
3	3.82,74.28	4.00,73.09	5.11,76.24	5.19,74.63	3.72,76.78	3.91	3.91
4	3.79,79.30	3.84,79.36	4.05,75.97	4.10,74.65	3.70,81.30	4.05
5	3.57,77.84	3.59,77.91	3.65,75.65	3.72,72.47	3.66,72.49	3.62	3.54
6	3.92,63.31	3.89,63.18	3.90,63.34	3.94	3.91,63.12	3.94	3.88
6'	3.76	3.79	3.75	3.78
Linkage	G/M/M₂/M₃/ $M 2 3$ /M_α	G/M	G/M	M/M₂/M₃	G/M/M₂/M₃/M_β

Fig.2 Probable structure of SDOP Note:M: β-D-Man; G: β-D-Gle; A: O-acetyl

References 23

[1]	Lü G.Y., Yan M. Q., Chen S. H., China Journal of Chinese Materia Medica, 2013, 38(4), 489—493
	(吕圭源, 颜美秋, 陈素红. 中国中药杂志, 2013, 38(4), 489—493)
[2]	Huang J., Zhang Y., Yan J., Xiao R., Xia B.H., Wu P., Lin L. M., Liao D. F., Chinese Pharmaceutical Journal, 2017, 52(7), 548—552
	(黄杰, 章莹, 严婧, 肖榕, 夏伯候, 吴萍, 林丽美, 廖端芳. 中国药学杂志, 2017, 52(7), 548—552)
[3]	Li L., Deng X.L., Zhao X. B., Zeng Y., Ouyang D. S., Anti-tumor Pharmacy, 2011, 1(2),90—94
	(李玲, 邓晓兰, 赵兴兵, 曾勇, 欧阳冬生. 肿瘤药学, 2011, 1(2), 90—94)
[4]	Tu G.C., Strait Pharmaceutical Journal, 2010, 22(2), 70—71(屠国昌. 海峡药学, 2010, 22(2), 70—71)
[5]	Cai H.L., Huang X. J., Nie S. P., Xie M. Y., Phillips G. O., Steve W. C., Bioactive Carbohydratesand Dietary Fibre, 2015, 5, 99—105
[6]	He Y.F., Liu W. L., Su R., Xiu Y., Pei J., Chem. Res. Chinese Universities, 2017, 33(2), 172—178
[7]	Chen Q., Lin J.H., Zhao D., Zhu X., Chem. Res. Chinese Universities, 2016, 32(5), 792—796
[8]	Hu X.C., Guo Y. B., Xiang J. S., Du Y. P., Journal of Anhui Agricultural Sciences, 2015, 43(15), 78—80, 84
	(胡贤春, 郭永兵, 向劲松, 杜永鹏. 安徽农业科学, 2015, 43(15), 78—80, 84)
[9]	Meng L.Z., Lv G. P., Hu D. J., Cheong K. L., Xie J., Zhao J., Li S. P., Molecules, 2013, 18(5), 5779—5791
[10]	Pan L.H., Li X. F., Wang M. N., Zha X. Q., Yang X. F., Liu Z. J., Luo Y. B., Luo J. P., Internation Journal of Biological Macromolecules, 2014, 64, 420—427
[11]	Nie S.P., Cai H. L., Food Science, 2012, 33(23), 356—361
	(聂少平, 蔡海兰. 食品科学, 2012, 33(23), 356—361)
[12]	He T.B., Huang Y. P., Yang L., Liu T. T., Gong W. Y., Wang X. J., Sheng J., Hu J. M., International Journal of Biological Macromolecules, 2016, 83(10), 34—41
[13]	Xia L.J., Liu X. F., Guo H. Y., Zhang H., Zhu J., Ren F. Z., Journal of Functional Foods, 2012, 4(1), 294—301
[14]	Liu Y., Yin L., Gong G.P., Peng Y. F., Huang L. J., Wang Z. F., Chem. [J]. Chinese Universities, 2016, 37(2), 261—268
	(刘洋, 殷璐, 龚桂萍, 彭艺芳, 黄琳娟, 王仲孚. 高等学校化学学报, 2016, 37(2), 261—268)
[15]	Guo X., Hu X.L., Wang Y. R., Yang Z. F., Wang Y. T., Li Z. T., Hu P., Chinese Traditional And Herbal Drugs, 2016, 47(9), 1508—1514(国欣, 胡小龙, 王月荣, 杨子峰, 王玉涛, 李征途, 胡坪. 中草药, 2016, 47(9), 1508—1514)
[16]	Luo B.J., Su W. M., Liu Y., China Rural of Science and Technology, 2006, (12), 38—39
	(罗秉俊, 苏卫明, 刘娅. 中国农村科技, 2006, (12), 38—39)
[17]	Needs P.W., Selvendran R. R., Carbohydrate Research, 1993, 245, 1—10
[18]	Xing X.H., Steve W. C., Nie S. P., Glyn O., Phillips H., Douglas G., Wang Q., Bioactive Carbohydrates And Dietary Fibre, 2014, 4(1), 74—83
[19]	Wang Z., Fang J.N., Chinese [J]. Anal. Chem., 2000, 28(2), 240—247
	(王展, 方积年. 分析化学, 2000, 28(2), 240—247)
[20]	Brink V.D. J., Vries D. R. P., Applied Microbiology And Biotechnology, 2011, 91(6), 1477—1492
[21]	Xing X.H., Cui S. W., Nie S. P., Phillips G. O., Goff H. D., Wang Q., Carbohydrate Polymers, 2015, 117, 422—433
[22]	Li B., Chen H.H., Xu S. Y., Natural Product Research and Development, 2005, 17(4), 523—526
	(李波, 陈海华, 许时婴. 天然产物研究与开发, 2005, 17(4), 523—526)
[23]	Liu L., Li K., Hao X., Wang G.Z., Qin X. M., Du G. H., Zhang X., Chem. [J]. Chinese Universities, 2016, 37(12), 2168—2175
	(刘磊, 李科, 郝霞, 王桂臻, 秦雪梅, 杜冠华, 张翔. 高等学校化学学报, 2016, 37(12), 2168—2175)