GC-MS分析牛樟芝多糖的单糖组成及其抗氧化活性
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摘要
研究牛樟芝的单糖组成和多糖的体外抗氧化活性。采用水提醇沉法提取牛樟芝多糖。以葡萄糖为标准品,苯酚硫酸法测定多糖的总糖含量。多糖经三氟乙酸水解后,糖腈乙酸酯法进行衍生化,采用气相色谱法-质谱法联用(GC-MS)分析测定单糖组成。通过DPPH自由基、ABTS自由基、OH自由基的清除能力评价牛樟芝多糖的体外抗氧化能力。结果表明,牛樟芝多糖的得率为3.02%±0.09%,总糖(以葡萄糖计)含量为55.42%±2.13%,单糖组成为半乳糖、葡萄糖、甘露糖,摩尔比为1∶7.14∶0.96。体外抗氧化实验结果表明牛樟芝多糖对DPPH自由基,ABTS自由基和OH自由基清除的IC_(50)分别为(0.584±0.008)、(1.182±0.058)、(1.384±0.133) mg/mL。GC-MS用于检测牛樟芝多糖的单糖组成简单、快速、灵敏,牛樟芝多糖对三种不同自由基具有一定的清除作用。
This study was conducted to investigate the monosaccharide composition and antioxidant activity of the polysaccharide from Antrodia cinnamomea(ACP). ACP was obtained with water extraction and ethanol precipitation method. The total sugar content of ACP was determined by using glucose as the standard substance and phenol-sulfuric acid method. ACP was hydrolyzed with trifluoroacetic acid,which monosaccharide compositions were analyzed by glyc-nitrile derivatization and gas chromatography-mass spectrometer(GC-MS)method. The antioxidant capacity of ACP in vitro was evaluated by DPPH,ABTS and OH free radicals scavenging assay. Results showed that total sugar contents of ACP was 55.42%±2.13% with the yield of 3.02%,mainly composed of galactose,glucose,mannose with an average molar ratio of 1∶7.14∶0.96,respectively. The result of antioxidant tests in vitro showed that ACP scavenged free radicals viz. DPPH free radical,ABTS free radical and OH free radical with IC_(50) values of(0.584±0.008),(1.182±0.058),(1.384±0.133) mg/mL,respectively. GC-MS method employed for analysis of the monosaccharide compositions of ACP was simple,rapid and sensitive. ACP exhibited a certain degree of scavenging activity for the three free radicals.
This study was conducted to investigate the monosaccharide composition and antioxidant activity of the polysaccharide from Antrodia cinnamomea(ACP). ACP was obtained with water extraction and ethanol precipitation method. The total sugar content of ACP was determined by using glucose as the standard substance and phenol-sulfuric acid method. ACP was hydrolyzed with trifluoroacetic acid,which monosaccharide compositions were analyzed by glyc-nitrile derivatization and gas chromatography-mass spectrometer(GC-MS)method. The antioxidant capacity of ACP in vitro was evaluated by DPPH,ABTS and OH free radicals scavenging assay. Results showed that total sugar contents of ACP was 55.42%±2.13% with the yield of 3.02%,mainly composed of galactose,glucose,mannose with an average molar ratio of 1∶7.14∶0.96,respectively. The result of antioxidant tests in vitro showed that ACP scavenged free radicals viz. DPPH free radical,ABTS free radical and OH free radical with IC_(50) values of(0.584±0.008),(1.182±0.058),(1.384±0.133) mg/mL,respectively. GC-MS method employed for analysis of the monosaccharide compositions of ACP was simple,rapid and sensitive. ACP exhibited a certain degree of scavenging activity for the three free radicals.
引文
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[19]龚劲松,袁峰,袁兵兵,等.高压均质法提取药用真菌多糖的研究[J].生物学杂志,2018,35(4):29-33.
[20]邓勇,张杰良,王兰英,等.李绍平.薄层色谱法分析不同虫草多糖的单糖组成[J].药物分析杂志,2018,38(1):13-21.
[21]Wang G J,Lin S Y,Wu W C,et al.DPPH radical scavenging and semicarbazide-sensitive amine oxidase inhibitory and cytotoxic activities of Taiwanofungus camphoratus(Chang-chih)[J].Bioscience Biotechnology and Biochemistry,2007,71(8):1873-1878.
[22]陈杰,董扬,鲁吉珂,等.3种不同产地灵芝子实体粗多糖体外抗氧化活性比较研究[J].食品工业科技,2016,37(21):100-104.
[23]张志军,李淑芬,魏雪生.灵芝多糖清除自由基活性的研究[J].食品研究与开发,2012,33(3):167-169.
[24]李绍平,吴定涛,赵静.糖谱及其在中药多糖质量控制中的应用[J].中国中药杂志,2015,40(17):3505-3513.
[2]张远腾,李晓波.牛樟芝化学成分及其药理作用研究进展[J].中草药,2016,47(6):1034-1042.
[3]Su Y K,Shih P H,Lee W H,et al.Antrodia cinnamomea sensitizes radio-/chemo-therapy of cancer stem-like cells by modulating micro RNA expression[J].Journal of Ethnopharmacology,2017,207:47-56.
[4]Hseu Y C,Huang H C,Hsiang C Y.Antrodia camphorata suppresses lipopolysaccharide-induced nuclear factor-kappaB activation in transgenic mice evaluated by bioluminescence imaging[J].Food and Chemical Toxicology,2010,48(8-9):2319-2325.
[5]Huang T T,Lan Y W,Ko Y F,et al.Antrodia cinnamomea produces anti-angiogenic effects by inhibiting the VEGFR2 signaling pathway[J].Journal of Ethnopharmacology.2018,220:239-249.
[6]Cheng P C,Hsu C Y,Chen C C,et al.In vivo immunomodulatory effects of Antrodia camphorata polysaccharides in a T1/T2 doubly transgenic mouse model for inhibiting infection of Schistosoma mansoni[J].Toxicology and Applied Pharmacology,2008,227(2):291-298.
[7]Liu Y G,Yang A H,Qu Y D,et al.Ameliorative effects of Antrodia cinnamomea polysaccharides against cyclophosphamide-induced immunosuppression related to Nrf2/HO-1 signaling in BALB/c mice[J].International Journal of BiologicalMacromolecules,2018,116:8-15.
[8]Chang C J,Lu C C,Lin C S,et al.Antrodia cinnamomea reduces obesity and modulates the gut microbiota in high-fat diet-fed mice[J].International Journal of Obesity,2018,42(2):231-243.
[9]Li Z W,Kuang Y,Tang S N,et al.Hepatoprotective activities of Antrodia camphorata and its triterpenoid compounds against CCl4-induced liver injury in mice[J].Journal of Ethnopharmacology,2017,206:31-39.
[10]郭元亨,张利军,曹丽丽,等.植物多糖中单糖组成分析技术的研究进展[J].食品科学,2018,39(1):326-332.
[11]张丽芝.单糖组成分析方法的研究进展[J].微生物学免疫学进展,2013,41(1):77-81.
[12]Su C H,Lai M N,Lin C C,et al Comparative characterization of physicochemical properties and bioactivities of polysaccharides from selected medicinal mushrooms[J].Applied Microbiology and Biotecnology,2016(100):4385-4393.
[13]张荔菲,刘莹,王益,等.超声辅助酶法提取柚子皮多糖工艺的响应面优化[J].食品工业科技,2018,39(5):146-150.
[14]王文洁,唐炜,俞玲娜,等.蒽酮-硫酸法与苯酚-硫酸法测定凉粉草多糖的比较[J].食品科技,2017,42(9):274-279.
[15]刘存芳,史娟,庞海霞,等.中药材野菊茎叶多糖的单糖组成及其抗氧化活性[J].食品科技,2018,43(5):191-197.
[16]Song Y,Wei X Q,Li M Y,et al.Nutritional composition and antioxidant properties of the fruits of a Chinese wild Passiflora foetida[J].Molecules,2018,23(2):459.
[17]Zhang X L,Gao H,Wang C Y,et al.Characterization and comparison of the structure and antioxidant activity of glycosylated whey peptides from two pathways[J].Food Chemistry,2018,257:279-288.
[18]Dadwal V,Agrawal H,Sonkhla K,et al.Characterization of phenolics,amino acids,fatty acids and antioxidant activity in pulp and seeds of high altitude Himalayan crab apple fruits(Malus baccata)[J].Journal of Food Science and Technology,2018,55(6):2160-2169.
[19]龚劲松,袁峰,袁兵兵,等.高压均质法提取药用真菌多糖的研究[J].生物学杂志,2018,35(4):29-33.
[20]邓勇,张杰良,王兰英,等.李绍平.薄层色谱法分析不同虫草多糖的单糖组成[J].药物分析杂志,2018,38(1):13-21.
[21]Wang G J,Lin S Y,Wu W C,et al.DPPH radical scavenging and semicarbazide-sensitive amine oxidase inhibitory and cytotoxic activities of Taiwanofungus camphoratus(Chang-chih)[J].Bioscience Biotechnology and Biochemistry,2007,71(8):1873-1878.
[22]陈杰,董扬,鲁吉珂,等.3种不同产地灵芝子实体粗多糖体外抗氧化活性比较研究[J].食品工业科技,2016,37(21):100-104.
[23]张志军,李淑芬,魏雪生.灵芝多糖清除自由基活性的研究[J].食品研究与开发,2012,33(3):167-169.
[24]李绍平,吴定涛,赵静.糖谱及其在中药多糖质量控制中的应用[J].中国中药杂志,2015,40(17):3505-3513.
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