新疆一枝蒿多糖的提取、纯化及其结构分析

doi:10.6048/j.issn.1001-4330.2018.06.012

摘要/Abstract

摘要： 【目的】 采用响应面法优化超声波法提取新疆一枝蒿多糖(ARP)的工艺条件,对其进行分离纯化和结构分析,这为ARP的结构分析及药理活性研究奠定基础。【方法】以新疆一枝蒿为原料,采用超声波法提取,Sevag法除蛋白,经DEAE纤维素-52柱和Sephadex G-100凝胶色谱柱纯化得ARP。利用紫外光谱扫描、红外光谱法对其化学结构进行分析。【结果】提取ARP的优化工艺条件为超声功率716 W、液料比36∶1和超声时间25 min,在此条件下,ARP的得率为3.92%±0.065%。ARP-2的红外吸收峰出现在917、1 100、1 147、1 332、1 423、1 611、1 742、2 939和3 410 cm^-1。【结论】优化的ARP制备工艺合理、可行,紫外光谱扫描结果显示ARP-2无明显的核酸和蛋白质吸收峰,ARP-2含有多糖类物质的特征吸收峰。

关键词: 新疆一枝蒿; 多糖; 超声波法; 纯化; 响应面法

Abstract: 【Objective】 The extraction of Artemisia rupestris L. polysaccharides (ARP) from Xinjiang was optimized by response surface method, and its separation, purification and preliminary structure analysis of ARP were carried out.【Method】Using Artemisia rupestris L. in Xinjiang as the raw material, ARP were obtained through ultrasonic-assisted method, Sevag deproteinization, DEAE cellulose and Sephadex G-100 gel filter column chromatographies. The structure of the ARP was analyzed and characterized by ultraviolet (UV) and infrared (IR) spectroscopy.【Result】A yield of ARP of 3.92%± 0.065% was obtained under optimized process condition (ultrasonic power 716 W, ratio of water to raw material 36 and ultrasonic time 25 min). The crude polysaccharides were purifed by fltration, DEAE-52 cellulose column and Sephadex G-100 size-exclusion chromatography in that order. A fraction, ARP-2 was obtained through the extraction and purifcation steps. The absorption spectra of the polysaccharides showed that the polysaccharides-related absorption peaks at 917, 1,100, 1,147, 1,332, 1,423, 1,611, 1,742, 2,939 and 3,410 cm^-1.【Conclusion】The optimized preparation process of ARP is reasonable and feasible. UV scanning showed that ARP-2 had no apparent absorption peaks of nucleic acid and protein. The characteristic absorption peak of ARP-2 containing polysaccharides was obtained by IR analysis, which has laid a foundation for the structural analysis and the pharmacological activity of ARP.

Key words: Artemisia rupestris L.; polysaccharides; ultrasonic extraction; purifcation; response surface methodology

中图分类号:

:S188

王旭辉, 徐鑫, 王卉, 叶凯, 涂振东, 晁群芳. 新疆一枝蒿多糖的提取、纯化及其结构分析[J]. 新疆农业科学, 2018, 55(6): 1085-1097.

WANG Xu-hui^1,2, XU Xin ³, WANG Hui², YE Kai², TU Zhen-dong², CHAO Qun-fang¹. Optimization, Purifcation and Preliminary Structural Characterization of Artemisia rupestris L. Polysaccharides[J]. Xinjiang Agricultural Sciences, 2018, 55(6): 1085-1097.

参考文献

[1] Ma, Y., Aisha, H. A., Liao, L., Aibai, S., Zhang, T., & Ito, Y. (2005). Preparative isolation and purification of rupestonic acid from the chinese medicinal plant artemisia rupestris, l. by high-speed counter-current chromatography. Journal of Chromatography A, 1076(1): 198-201.
[2] Forabosco, A., Bruno, G., Sparapano, L., Liut, G., Marino, D., & Delben, F. (2006). Pullulans produced by strains of cryphonectria parasitica-i. production and characterisation of the exopolysaccharides. Carbohydrate Polymers, 63(4): 535-544.
[3] Yang, B., Jiang, Y., Zhao, M., Chen, F., Wang, R., & Chen, Y., et al. (2009). Structural characterisation of polysaccharides purified from longan (dimocarpus longan lour.) fruit pericarp. Food Chemistry, 115(2): 609-614.
[4] Feng, K., Chen, W., Sun, L., Liu, J., Zhao, Y., & Li, L., et al. (2015). Optimization extraction, preliminary characterization and antioxidant activity in vitro, of polysaccharides from stachys sieboldii, miq. tubers. Carbohydr Polym, 125(10):45-52.
[5] Zheng, Q., Ren, D., Yang, N., & Yang, X. (2016). Optimization for ultrasound-assisted extraction of polysaccharides with chemical composition and antioxidant activity from the artemisia sphaerocephala krasch seeds. International Journal of Biological Macromolecules, (91): 856-866.
[6] Ferreira, I. C., Heleno, S. A., Reis, F. S., Stojkovic, D., Queiroz, M. J., & Vasconcelos, M. H., et al. (2015). Chemical features of ganoderma polysaccharides with antioxidant, antitumor and antimicrobial activities. Phytochemistry, (114): 38-55.
[7] Errea, M. I., & Matulewicz, M. C. (1996). Hot water-soluble polysaccharides from tetrasporic pterocladia capillacea. Phytochemistry, 42(4): 1,071-1,073.
[8] Bendjeddou, D., Lalaoui, K., & Satta, D. (2003). Immunostimulating activity of the hot water-soluble polysaccharide extracts of anacyclus pyrethrum, alpinia galanga and citrullus colocynthis. Journal of Ethnopharmacology, 88(2): 155-160.
[9] Li, H., Pordesimo, L., & Weiss, J. (2004). High intensity ultrasound-assisted extraction of oil from soybeans. Food Research International, 37(7): 731-738.
[10] Vinatoru, M., Toma, M., Radu, O., Filip, P. I., Lazurca, D., & Mason, T. J. (1997). The use of ultrasound for the extraction of bioactive principles from plant materials. Ultrasonics Sonochemistry, 4(2): 135-139.
[11] Wang, Y., Cheng, Z., Mao, J., Fan, M., & Wu, X. (2009). Optimization of ultrasonic-assisted extraction process of poria cocos polysaccharides by response surface methodology. Carbohydrate Polymers, 77(4):713-717.
[12] Entezari, M. H., Nazary, S. H., & Khodaparast, M. H. H. (2004). The direct effect of ultrasound on the extraction of date syrup and its micro-organisms. Ultrasonics Sonochemistry, 11(6):379-384.
[13] Vilkhu, K., Mawson, R., Simons, L., & Bates, D. (2008). Applications and opportunities for ultrasound assisted extraction in the food industry - a review. Innovative Food Science & Emerging Technologies, 9(2): 161-169.
[14] G. E. P. Box, & K. B. Wilson. (1951). On the experimental attainment of optimum conditions. Journal of the Royal Statistical Society. Series B (Methodological) , 13(1): 1-45.
[15] Giovanni, M. (1983). Response surface methodology and product optimization. Food Technology, 37(11): 41-45.
[16] enol banolu, & Ainsworth, P. (2004). Effect of canning on the starch gelatinization and protein in vitro digestibility of tarhana, a wheat flour-based mixture. Journal of Food Engineering, 64(2): 243-247.
[17] Liang, R. J. (2008). Optimization of extraction process of glycyrrhiza glabra polysaccharides by response surface methodology. Carbohydrate Polymers, 74(4): 858-861.
[18] Varnalis, A. I., Brennan, J. G., Macdougall, D. B., & Gilmour, S. G. (2004). Optimisation of high temperature puffing of potato cubes using response surface methodology. Journal of Food Engineering, 61(2): 153-163.
[19] Vega, P. J., Balaban, M. O., Sims, C. A., O'Keefe, S. F., & Cornell, J. A. (2010). Supercritical carbon dioxide extraction efficiency for carotenes from carrots by rsm. Journal of Food Science, 61(4): 757-759.
[20] Xu, H., Sun, L. P., Shi, Y. Z., Wu, Y. H., Zhang, B., & Zhao, D. Q. (2008). Optimization of cultivation conditions for extracellular polysaccharide and mycelium biomass by morchella esculenta, as51620. Biochemical Engineering Journal, 39(1): 66-73.
[21] Yan, Y. L., Yu, C. H., Chen, J., Li, X. X., Wang, W., & Li, S. Q. (2011). Ultrasonic-assisted extraction optimized by response surface methodology, chemical composition and antioxidant activity of polysaccharides from tremella mesenterica. Carbohydrate Polymers,83(1): 217-224.
[22] Zhao, Q., Kennedy, J. F., Wang, X., Yuan, X., Zhao, B., & Peng, Y., et al. (2011). Optimization of ultrasonic circulating extraction of polysaccharides from asparagus officinalis using response surface methodology. International Journal of Biological Macromolecules, 49(2):181-187.
[23] Yang, W., Fang, Y., Liang, J., & Hu, Q. (2011). Optimization of ultrasonic extraction of flammulina velutipes polysaccharides and evaluation of its acetylcholinesterase inhibitory activity. Food Research International, 44(5):1,269-1,275.
[24] Zykwinska, A., Rondeau-Mouro, C., Garnier, C., Thibault, J. F., & Ralet, M. C. (2006). Alkaline extractability of pectic arabinan and galactan and their mobility in sugar beet and potato cell walls. Carbohydrate Polymers,65(4): 510-520.
[25] Ye, C. L., & Jiang, C. J. (2011). Optimization of extraction process of crude polysaccharides from plantago asiatica l. by response surface methodology. Carbohydrate Polymers, 84(1): 495-502.
[26] L. Navarini, R. Gilli, V. Gombac, A. Abatangelo, M. Bosco, & R. Toffanin. (1999). Polysaccharides from hot water extracts of roasted coffea arabica, beans: isolation and characterization. Carbohydrate Polymers,40(1): 71-81.
[27] Copeland, K. A. F. (2001). Design and analysis of experiments, 5th ed. Journal of Quality Technology, 33(3): 382-383.
[28] RV Muralidhar, & RR Chirumamila. (2001). A response surface approach for the comparison of lipase production by canida cylindracea using two different carbon sources. Biochemical Engineering Journal, 9(1):17-23.
[29] Muralidhar, R. V., Chirumamilla, R. R., Ramachandran, V. N., Marchant, R., & Nigam, P. (2002). Resolution of -proglumide using lipase from candida cylindraceae. Cheminform, 10(5):1,471-1,475.
[30] Li, J., Fan, L., & Ding, S. (2011). Isolation, purification and structure of a new water-soluble polysaccharide from zizyphus jujuba cv. jinsixiaozao. Carbohydrate Polymers, 83(2):477-482.