酸角果肉多糖的提取工艺优化与初级结构分析
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摘要
以酸角果肉为原料,利用响应面分析法改进超声波辅助提取酸角果肉多糖(polysaccharides from Tamarindus indica L. Pulp fruit,PFTP)的条件参数。基于单因素试验结果,设置自变量为提取时间、超声功率、提取温度与液料比,响应值为酸角果肉多糖得率,进行四因素三水平的响应面分析。获得最佳提取参数:提取时间40 min、超声功率200 W、提取温度50℃、液料比30∶1(mL/g),此时得率为31.94%,高于文献优化条件下的热水浸提得率23.44%。结合紫外吸收光谱、红外吸收光谱与高效阴离子交换色谱法分析酸角果肉多糖的初级结构,紫外吸收光谱表明酸角果肉多糖含有少量蛋白质,无核酸;红外吸收光谱表明酸角果肉多糖出现多糖的特征吸收,具有β-糖苷键结构;高效阴离子交换色谱图表明酸角果肉多糖的主要单糖组成为阿拉伯糖、半乳糖、鼠李糖、岩藻糖、木糖、葡萄糖和半乳糖醛酸,对应的摩尔比为45.56∶15.67∶6.41∶1∶5.14∶14.27∶1.95,高效排阻色谱法测得酸角果肉多糖的分子量为8.262×10~5g/mol,且分布均一,其半乳糖醛酸含量为30.02%。
Response surface methodology(RSM) was used to optimize ultrasonic extraction process of polysaccharides from Tamarindus indica L. Pulp fruit(PFTP). In single factor experiments,extraction time,ultrasonic power,extraction temperature and material solution ratio were used as independent variables and tested at three levels,while the polysaccharide yield was the response variable. The obtained optimum conditions were listed as follows:extraction time(40 min),ultrasonic power(200 W),extraction temperature(50 ℃),and the material solution ratio(1 ∶ 30 g/mL). Under the optimized conditions,the yield was 31.94 %,higher than the reported value(23.44 %)which was obtained by thermal water extraction. Primary structure of the polysaccharide was determined by ultraviolet(UV)absorption spectrum,infrared(IR)absorption spectrum and high-performance anion-exchange chromatography(HPAEC),suggested that polysaccharides from PFTP contained protein but without nucleic acid. IR showed PFTP had characteristic absorption peaks of polysaccharide and β-glycosidic bond. PFTP were mainly composed of seven monosaccharide units,including arabinose,galactose,rhamnose,fucose,xylose,glucose and galacturonic acid with a molar ratio of 45.56 ∶ 15.67 ∶ 6.41 ∶ 1 ∶5.14 ∶ 14.27 ∶ 1.95. The relative molecular weight of the PFTP was 8.262×10~5 g/mol and the distribution was uniform by high performance size-exclusion chromatography(HPSEC). The polysaccharide showed 30.02 % of uronic acid.
Response surface methodology(RSM) was used to optimize ultrasonic extraction process of polysaccharides from Tamarindus indica L. Pulp fruit(PFTP). In single factor experiments,extraction time,ultrasonic power,extraction temperature and material solution ratio were used as independent variables and tested at three levels,while the polysaccharide yield was the response variable. The obtained optimum conditions were listed as follows:extraction time(40 min),ultrasonic power(200 W),extraction temperature(50 ℃),and the material solution ratio(1 ∶ 30 g/mL). Under the optimized conditions,the yield was 31.94 %,higher than the reported value(23.44 %)which was obtained by thermal water extraction. Primary structure of the polysaccharide was determined by ultraviolet(UV)absorption spectrum,infrared(IR)absorption spectrum and high-performance anion-exchange chromatography(HPAEC),suggested that polysaccharides from PFTP contained protein but without nucleic acid. IR showed PFTP had characteristic absorption peaks of polysaccharide and β-glycosidic bond. PFTP were mainly composed of seven monosaccharide units,including arabinose,galactose,rhamnose,fucose,xylose,glucose and galacturonic acid with a molar ratio of 45.56 ∶ 15.67 ∶ 6.41 ∶ 1 ∶5.14 ∶ 14.27 ∶ 1.95. The relative molecular weight of the PFTP was 8.262×10~5 g/mol and the distribution was uniform by high performance size-exclusion chromatography(HPSEC). The polysaccharide showed 30.02 % of uronic acid.
引文
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[27]王海燕,戴军,陈尚卫.灵芝菌丝体多糖的分离纯化及其单糖组成分析与分子量测定[J].食品与机械,2015(5):201-205
[2]李维熙,王葳,杨柏荣,等.酸角的化学成分及生物活性研究现状[J].国际药学研究杂志,2016,43(4):697-704
[3]赵一鹤,杨时宇,李昆.世界酸角研究现状及进展[J].云南农业大学学报(自然科学),2005,20(1):65-72
[4]吉宏武,丁霄霖.罗望子研究进展[J].中国野生植物资源,2000(6):10-14
[5]Shrikant Hiwale.Sustainable Horticulture in Semiarid Dry Lands[M].Berlin:Springer,2015
[6]Makkawi N A,Kinattingal N,Manjula S N,et al.CARDIOPRO-TECTIVE EFFECT OF TAMARINDUS INDICA LINN AGAINSTISOPROTERENOL INDUCED MYOCARDIAL INFARCTION INRATS[J].International Journal of Pharmacy&Pharmaceutical Sciences,2016,8(5):254-260
[7]Gupta C,Prakash D,Gupta S.Studies on the antimicrobial activity of tamarind(Tamarindus indica)and its potential as food biopreservative.[J].International Food Research Journal,2014,21(6):2437-2441
[8]Maiti R,Jana D,Das U K,et al.Antidiabetic effect of aqueous extract of seed of Tamarindus indica in streptozotocin-induced diabetic rats[J].Journal of Ethnopharmacology,2004,92(1):85-91
[9]王玲.罗望子有效成分的提取纯化及抗氧化作用研究[D].长沙:湖南农业大学,2013:19-31
[10]Vilkhu K,Mawson R,Simons L,et al.Applications and opportunities for ultrasound assisted extraction in the food industry-A review[J].Innovative Food Science&Emerging Technologies,2008,9(2):161-169
[11]Hong S,Pan L H,Luo J P.Progress of ultrasound-assisted exaction of biological polysaccharides[J].Science&Technology of Food Industry,2011,32(8):481-484
[12]Yan W,Cui S W,Jian T,et al.Optimization of extraction process of crude polysaccharides from boat-fruited sterculia seeds by response surface methodology[J].Food Chemistry,2007,105(4):1599-1605
[13]戴军,梁立娜,尹鸿萍,等.高效阴离子交换色谱法分析杜氏盐藻多糖的单糖组成[J].食品与发酵工业,2006,32(11):131-135
[14]郭欣,高向东,杨晓兵.酸性多糖中的葡萄糖醛酸与中性糖的含量测定[J].中国生化药物杂志,2004,25(2):100-101
[15]贾媛颖.橄榄多糖提取、分离及性质研究[D].福州:福建农林大学,2012:15-39
[16]张惟杰.糖复合物生化研究技术[M].杭州:浙江大学出版社,1999
[17]Aguiló-Aguayo I,Walton J,Vi觡as I,et al.Ultrasound assisted extraction of polysaccharides from mushroom by-products[J].LWT-Food Science and Technology,2017,77:92-99
[18]Maran J P,Manikandan S,Nivetha C V,et al.Ultrasound assisted extraction of bioactive compounds from Nephelium lappaceum,L.fruit peel using central composite face centered response surface design[J].Arabian Journal of Chemistry,2017,10:S1145-S1157
[19]Zhu C P,Zhai X C,Li L Q,et al.Response surface optimization of ultrasound-assisted polysaccharides extraction from pomegranate peel.[J].Food Chemistry,2015,177:139-146
[20]Chen R,Li S,Liu C,et al.Ultrasound complex enzymes assisted extraction and biochemical activities of polysaccharides from Epimedium leaves[J].Process Biochemistry,2012,47(12):2040-2050
[21]Chen C,You L J,Abbasi A M,et al.Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro[J].Carbohydrate Polymers,2015,130:122-132
[22]Ying Z,Han X,Li J.Ultrasound-assisted extraction of polysaccharides from mulberry leaves[J].Food Chemistry,2011,127(3):1273-1279
[23]Maran J P,Priya B.Ultrasound-assisted extraction of polysaccharide from Nephelium lappaceum L.fruit peel.[J].International Journal of Biological Macromolecules,2014,70(8):530-536
[24]王琪,李文亚.晋西北酸粥发酵工艺的研究[J].食品与发酵工业,2017,43(9):137-143
[25]Faurot A L,Saulnier L,Berot S,et al.Large scale isolation of watersoluble and water-insoluble pentosans from wheat flour.[J].LWT-Food Science and Technology,1995,28(4):436-441
[26]吴艳,顾小红,Steve W Cui,等.胖大海酸性多糖的分离纯化及初步结构研究[J].食品研究与开发,2006,27(7):95-98
[27]王海燕,戴军,陈尚卫.灵芝菌丝体多糖的分离纯化及其单糖组成分析与分子量测定[J].食品与机械,2015(5):201-205