The analysis of enzymatic products of Ganoderma atrum polysaccharides by pre-column derivatization HPLC-UV-Q-TOF/MS
-
摘要: 采用1-苯基-3-甲基-5-吡唑啉酮(PMP)对灵芝粗多糖酶解产物进行衍生,并利用液相-紫外检测器-高分辨四级杆串联飞行时间质谱法(HPLC-UV-Q-TOF-MS)检测分析灵芝多糖的单糖组成及其酶解产物。结果表明,灵芝多糖的单糖组成主要是葡萄糖、甘露糖和半乳糖、及少量的戊糖、岩藻糖和葡萄糖醛酸。经HPLC-Q-TOF-MS分析鉴定,β-葡聚糖酶的降解产物为单糖、二糖到五糖的低聚糖,证明灵芝多糖多存在β-糖苷键,而果胶复合酶将黑灵芝多糖主要降解成单糖。Abstract: 1-phenyl-3- methyl-5- pyrazolone ( PMP) was applied to derivate polysaccharide degradation product.Then liquid chromatography with ultraviolet detector- high resolution quadrupole tandem time of flight mass spectrometry ( HPLC- UV- Q- TOF- MS) was used to detect and analyze monosaccharide compositions and enzymatic hydrolysate of Ganoderma atrum polysaccharide. The results showed that the monosaccharide compositions of Ganoderma atrum polysaccharides were mainly glucose, mannose, galactose. The degradation products of β-glucanase were monosaccharides and oligosaccharides from disaccharide to pentasaccharides, but Ganoderma atrum polysaccharides were mainly degraded into monosaccharides by pectic enzyme.
-
[1] 陈奕, 谢明勇, 弓晓峰.黑灵芝提取物清除DPPH自由基的作用[J].天然产物研究与开发, 2006, 18:917-921. [2] 毛健, 马海乐.灵芝多糖的研究进展[J].食品科学, 2010, 31 (1) :295-299. [3] Xu Zeng-tao, Chen Xiu-ping, ZHONG Zhang-feng, et al.Ganoderma lucidum Polysaccharides:Immunomodulation and Potential Anti-Tumor Activities[J].The American Journal of Chinese Medicine, 2011, 39 (1) :15-27.
[4] 余强, 聂少平, 李文娟, 等.黑灵芝多糖对D-半乳糖致衰老小鼠的作用研究[J].食品科学, 2009, 30 (17) :305-307. [5] 王本祥.现代中药药理与临床[M].天津:天津科技翻译出版公司, 2004:744-752. [6] 何晋浙, 邵平, 孟祥河, 等.灵芝多糖的结构特征分析[J].分析化学研究报告, 2010, 38 (3) :372-376. [7] 罗璐.灵芝多糖的结构分析与功能特性的研究[D].武汉:华中农业大学, 2009. [8] Zhang Hui, Li Wen-juan, Nie Shao-ping, et al.Structural characterisation of a novel bioactive polysaccharide from Ganoderma atrum[J].Carbohydrate Polymers, 2012, 88:1047-1054.
[9] J Guan, S P Li.Discrimination of polysaccharides from traditional Chinese medicines using saccharide mappingEnzymatic digestion followed by chromatographic analysis[J].Journal of Pharmaceutical and Biomedical Analysis, 2010, 51:590-598.
[10] Xin Di, Kelvin K C Chan, Hei Wun Leung.Fingerprint profiling of acid hydrolyzates of polysaccharides extracted from the fruiting bodies and spores of Lingzhi by high-performance thinlayer chromatography[J].Journal of Chromatography A, 2003, 1018 (1) :85-95.
[11] 陈克克.地瓜儿多糖的单糖组成实验中两种流动相的比较研究[J].陕西农业科学, 2009 (5) :41-49. [12] 张汇, 鄢嫣, 聂少平, 等.黑灵芝不同部位多糖成分分析及抗氧化活性[J].食品科学, 2011, 32 (1) :56-61. [13] Chen Yi, Xie Mingyong, Nie Shaoping, et al.Purification, composition analysis and antioxidant activityof a polysaccharide from the fruiting bodies of Ganoderma atrum[J].Food Chemistry, 2008, 107:231-241.
[14] 魏晓飞, 王在贵, 陈键.β-葡聚糖酶的酶学性质研究[J].中国饲料, 2007 (9) :17-19. [15] 郭雨桐, 肖文军.β-葡聚糖酶降解茯苓多糖的效应[J].北方园艺, 2011 (24) :18-22. [16] 薛长湖, 张永勤.果胶及果胶酶研究进展[J].食品与生物技术学报, 2005, 24 (6) :94-99. [17] 汪江山, 赵欣捷, 郑育芳, 等.超高效液相色谱/飞行时间质谱用于人参皂甙Rg3作用后大鼠尿液代谢物指纹图谱分析及标记物的鉴定[J].色谱, 2006, 24 (1) :5-9. [18] 毛文君, 林洪, 管华诗.琼胶寡糖的ESI-MS分析研究[J].中国水产科学, 2001, 8 (3) :69-72. [19] Domon B, Costello C E.Structure elucidation of glycosph ingolipids and gangliosides using high-performance tandem mass spectrometry[J].Biochemistry, 1988, 27:1534-1543.
计量
- 文章访问数:
- HTML全文浏览量:
- PDF下载量: