油茶籽饼粕多糖的提取工艺优化及结构鉴定
|
摘要
[目的]以油茶籽饼粕为原料,优化油茶籽饼粕多糖提取工艺,并对纯化后的多糖进行结构鉴定。[方法]根据单因素试验结果,以超声时间10、15、20 min;料液比1∶15、1∶20、1∶25;浸提温度50、60、70℃;浸提时间60、90、120 min为影响因素,多糖得率为指标,通过正交试验设计优化提取工艺。粗多糖经脱色,脱蛋白处理后,再经过DEAE-52纤维素阴离子交换树脂纯化,并对纯化多糖组分进行紫外光谱和高效液相色谱分析。[结果]超声时间10 min,料液比1∶20,浸提温度60℃,浸提时间120 min为最佳工艺条件,此条件下油茶籽饼粕多糖的平均得率16.76%。粗多糖经脱色、脱蛋白、纤维素阴离子交换树脂柱层析得到纯化组分CCP。紫外光谱表明CCP为较纯的均一多糖组分,高效液相色谱表明CCP的单糖组成为甘露糖和葡萄糖,其物质的量比为:1.91∶1.00。同时还含有少量的鼠李糖。[结论]该工艺优化合理,多糖的结构得到了初步的确证。研究为油茶籽饼粕多糖的后续理化活性分析、结构表征提供了理论依据。
[Objective]Camellia oleifera seed cake was used as raw material to optimize the extraction process of polysaccharides and to analyze the structure of purified polysaccharide.[Method]Ultrasonic time( 10,15,20 min),ratio of material to liquid( 1 ∶15,1 ∶20,1 ∶25),extraction temperature( 50,60,70 ℃),extraction time( 60,90,120 min) were choosen as influencing factors on the basis of single factor experiments,extraction rate of polysaccharides was taken as evaluation indexe.Orthogonal test was used to optimize polysaccharide extraction process.The crude polysaccharides was decolorized,deproteinized and purified by DEAE-52 Cellulose anion exchange chromatography respectively.The purified polysaccharides were analyzed by UV and high performance liquid chromatography( HPLC).[Result]The best extraction process was ultrasonic time 10 min,ratio of material to liquid 1 ∶20,extraction temperature 60 ℃,extraction time 120 min.Under these conditions,the average extraction rate of polysaccharides reached 16.76%.Purified component was obtained and named as CCP.UV spectrum showed that CCP was a homogeneous polysaccharide component.HPLC showed that the purified polysaccharide was mainly composed of mannose,glucose with molar ratio of 1.91 ∶1.00,and tiny amounts of rhamnose.[Conclusion]The optimized process is reasonable and the structure of polysaccharides is preliminary identificated.It can provide a theoretical basis for further activity analysis and structural characterization of camellia oleifera seed cake polysaccharides.
[Objective]Camellia oleifera seed cake was used as raw material to optimize the extraction process of polysaccharides and to analyze the structure of purified polysaccharide.[Method]Ultrasonic time( 10,15,20 min),ratio of material to liquid( 1 ∶15,1 ∶20,1 ∶25),extraction temperature( 50,60,70 ℃),extraction time( 60,90,120 min) were choosen as influencing factors on the basis of single factor experiments,extraction rate of polysaccharides was taken as evaluation indexe.Orthogonal test was used to optimize polysaccharide extraction process.The crude polysaccharides was decolorized,deproteinized and purified by DEAE-52 Cellulose anion exchange chromatography respectively.The purified polysaccharides were analyzed by UV and high performance liquid chromatography( HPLC).[Result]The best extraction process was ultrasonic time 10 min,ratio of material to liquid 1 ∶20,extraction temperature 60 ℃,extraction time 120 min.Under these conditions,the average extraction rate of polysaccharides reached 16.76%.Purified component was obtained and named as CCP.UV spectrum showed that CCP was a homogeneous polysaccharide component.HPLC showed that the purified polysaccharide was mainly composed of mannose,glucose with molar ratio of 1.91 ∶1.00,and tiny amounts of rhamnose.[Conclusion]The optimized process is reasonable and the structure of polysaccharides is preliminary identificated.It can provide a theoretical basis for further activity analysis and structural characterization of camellia oleifera seed cake polysaccharides.
引文
[1]姚小华,王开良,罗细芳,等.我国油茶产业化现状及发展思路[J].林业科技开发,2005,19(1):3-6.
[2]刘小如,张丽美,胡蒋宁,等.油茶粕多糖的分级纯化及结构研究[J].食品科学,2013,34(23):96-102.
[3]王元凤,俞兰,魏新林.从油茶饼粕中提取油茶籽多糖和茶皂素的工艺研究[J].食品工业,2008(6):29-32.
[4]郭华,刁全平,张博,等.杨树口蘑多糖的超声波辅助提取工艺及其抗氧化活性[J].食品工业科技,2018(5):180-184.
[5]王胜男,赵玲玲,张红运,等.提取方法对大豆种皮多糖乳化能力的影响[J].中国食品学报,2018,18(1):154-161.
[6]吴宪玲,于晓红,刘涛,等.响应面法优化西洋参多糖的酶解辅助提取工艺[J].食品工业,2016,37(1):118-122.
[7]蔡锦源,梁莹,吴庆华,等.白及多糖的提取工艺及其生物活性研究[J].食品工业,2018(1):45-49.
[8]黎英,陈雪梅,严月萍,等.超声波辅助酶法提取红腰豆多糖工艺优化[J].农业工程学报,2015,31(15):293-301.
[9]CHEN Y,XIE M Y,NIE S P,et al.Purification,composition analysis and antioxidant activity of a polysaccharide from the fruiting bodies of Ganoderma atrum[J].Food chemistry,2008,107(1):231-241.
[10]LIU Y,GONG G P,SUN Y J,et al.Isolation,structural characterization,and immunological activity of a polysaccharide LRLP4-A from the leaves of Lycium ruthenicum[J].Journal of carbohydrate chemistry,2016,35(1):40-56.
[11]ZHENG Y F,ZHANG S,WANG Q,et al.Characterization and hypoglycemic activity of aβ-pyran polysaccharides from bamboo shoot(Leleba oldhami Nakal)shells[J].Carbohydrate polymers,2016,144:438-446.
[12]赵雪竹,刘洋,王艳萍,等.原料纯度对白及多糖疏水改性反应取代度的影响[J].药学实践杂志,2015,33(3):221-225.
[13]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(9):274-279.
[14]王琳炜,欧阳臻,张碧娟,等.霍山铁皮石斛多糖的脱蛋白工艺及结构分析[J].食品科学,2017,38(12):164-170.
[15]赵艳,王白娟,杨青松,等.红雪茶多糖过氧化氢脱色工艺优化[J].南方农业学报,2016,47(5):710-714.
[16]戴军,朱松,汤坚,等.PMP柱前衍生高效液相色谱法分析杜氏盐藻多糖的单糖组成[J].分析测试学报,2007,26(2):206-210.
[17]李薇,夏晴,孙成荣,等.柱前衍生(PMP)-HPLC法测定不同品种甘草多糖中单糖组成[J].辽宁中医药大学学报,2014,16(1):56-58.
[18]王向红,吉爽爽,生庆海,等.柱前衍生高效液相色谱法检测8种枣水溶性多糖的单糖组成[J].中国食品学报,2014,14(9):257-262.
[19]魏晓蕊.桑叶中1-脱氧野尻霉素的提取纯化及其降血糖活性的研究[D].扬州:扬州大学,2012.
[20]王强,李盛钰,杨帆,等.玉竹中性多糖的分离纯化及单糖组成分析[J].食品科学,2010,31(15):100-102.
[21]宋学伟,任磊,韩泳平,等.大花红景天多糖RCPS分离纯化及单糖组成分析[J].光谱学与光谱分析,2008,28(3):642-644.
[22]施君君.牡丹籽粕多糖理化特性及抗氧化、抗肿瘤功能研究[D].合肥:合肥工业大学,2017:12-13.
[23]林华娟,田晓春,秦小明,等.金花茶多糖单一成分的化学结构特征解析[J].食品科学,2013,34(3):141-146.
[2]刘小如,张丽美,胡蒋宁,等.油茶粕多糖的分级纯化及结构研究[J].食品科学,2013,34(23):96-102.
[3]王元凤,俞兰,魏新林.从油茶饼粕中提取油茶籽多糖和茶皂素的工艺研究[J].食品工业,2008(6):29-32.
[4]郭华,刁全平,张博,等.杨树口蘑多糖的超声波辅助提取工艺及其抗氧化活性[J].食品工业科技,2018(5):180-184.
[5]王胜男,赵玲玲,张红运,等.提取方法对大豆种皮多糖乳化能力的影响[J].中国食品学报,2018,18(1):154-161.
[6]吴宪玲,于晓红,刘涛,等.响应面法优化西洋参多糖的酶解辅助提取工艺[J].食品工业,2016,37(1):118-122.
[7]蔡锦源,梁莹,吴庆华,等.白及多糖的提取工艺及其生物活性研究[J].食品工业,2018(1):45-49.
[8]黎英,陈雪梅,严月萍,等.超声波辅助酶法提取红腰豆多糖工艺优化[J].农业工程学报,2015,31(15):293-301.
[9]CHEN Y,XIE M Y,NIE S P,et al.Purification,composition analysis and antioxidant activity of a polysaccharide from the fruiting bodies of Ganoderma atrum[J].Food chemistry,2008,107(1):231-241.
[10]LIU Y,GONG G P,SUN Y J,et al.Isolation,structural characterization,and immunological activity of a polysaccharide LRLP4-A from the leaves of Lycium ruthenicum[J].Journal of carbohydrate chemistry,2016,35(1):40-56.
[11]ZHENG Y F,ZHANG S,WANG Q,et al.Characterization and hypoglycemic activity of aβ-pyran polysaccharides from bamboo shoot(Leleba oldhami Nakal)shells[J].Carbohydrate polymers,2016,144:438-446.
[12]赵雪竹,刘洋,王艳萍,等.原料纯度对白及多糖疏水改性反应取代度的影响[J].药学实践杂志,2015,33(3):221-225.
[13]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(9):274-279.
[14]王琳炜,欧阳臻,张碧娟,等.霍山铁皮石斛多糖的脱蛋白工艺及结构分析[J].食品科学,2017,38(12):164-170.
[15]赵艳,王白娟,杨青松,等.红雪茶多糖过氧化氢脱色工艺优化[J].南方农业学报,2016,47(5):710-714.
[16]戴军,朱松,汤坚,等.PMP柱前衍生高效液相色谱法分析杜氏盐藻多糖的单糖组成[J].分析测试学报,2007,26(2):206-210.
[17]李薇,夏晴,孙成荣,等.柱前衍生(PMP)-HPLC法测定不同品种甘草多糖中单糖组成[J].辽宁中医药大学学报,2014,16(1):56-58.
[18]王向红,吉爽爽,生庆海,等.柱前衍生高效液相色谱法检测8种枣水溶性多糖的单糖组成[J].中国食品学报,2014,14(9):257-262.
[19]魏晓蕊.桑叶中1-脱氧野尻霉素的提取纯化及其降血糖活性的研究[D].扬州:扬州大学,2012.
[20]王强,李盛钰,杨帆,等.玉竹中性多糖的分离纯化及单糖组成分析[J].食品科学,2010,31(15):100-102.
[21]宋学伟,任磊,韩泳平,等.大花红景天多糖RCPS分离纯化及单糖组成分析[J].光谱学与光谱分析,2008,28(3):642-644.
[22]施君君.牡丹籽粕多糖理化特性及抗氧化、抗肿瘤功能研究[D].合肥:合肥工业大学,2017:12-13.
[23]林华娟,田晓春,秦小明,等.金花茶多糖单一成分的化学结构特征解析[J].食品科学,2013,34(3):141-146.