A Study on the Effect on an Ultrasound-enhanced Aqueous Two-
发布时间：2021-09-04 13:18
植物多糖是天然的,功能强大的,无细胞毒性的化合物,越来越多地被用作传统药物的替代品,用来减少因氧化应激所导致的慢性疾病。但是,由于大多数提取方法会使多糖遭到一定程度的破坏,从而影响其生物活性,因此在提取过程中提高其提取率和生物活性仍然是一个挑战。因此,一种利用多频超声辅助醇/盐水溶液两相萃取技术被开发和探索以用来提取和纯化黄芪和香菇多糖,其主要目的是提高产量并增强其抗氧化能力。通过以下方式实现了目标:（i）选择合适的双水相体系作为提取和纯化体系。（ii）通过响应面法优化提取参数,以提高多频超声中两相提取的性能。（iii）在脱盐过程中使用不同时间的超声波（10分钟,20分钟和30分钟）,使富含多糖和盐的底部相达到脱盐并纯化多糖的目的。最后,对脱盐提取物的理化,微观结构和体外抗氧化能力进行了研究。主要结果和内容如下:1.双频超声辅助乙醇/盐双水相萃取黄芪多糖的研究。本课题研究了双频超声辅助乙醇/盐双水相对黄芪多糖（APS）的萃取率和脱盐率的影响,并研究了其微观结构和抗氧化性能的影响。将提取物脱盐10(APS10),20(APS20)和30(APS...
【文章来源】：江苏大学江苏省
【文章页数】：124 页
【学位级别】：硕士
【文章目录】：
ACKNOWLEDGEMENT
ABSTRACT
摘要
ABBREVIATIONS
CHAPTER 1 INTRODUCTION AND RESEARCH OBJECTIVES
1.1 General background
1.2 Astragalus membranaceus
1.2.1 Composition
1.2.2 Health-related properties
1.3 Lentinus edodes
1.3.1 Composition
1.3.2 Health-related properties
1.4 Extraction of polysaccharides
1.5 Conventional polysaccharide extraction methods
1.5.1 Hot water extraction
1.5.2 Reflux extraction
1.5.3 Complex enzyme hydrolysis method
1.6 Novel polysaccharide extraction methods
1.6.1 Ultrasound-assisted extraction
1.6.1.1 Multi-frequency ultrasound
1.6.2 Subcritical water extraction(SWE)
1.6.3 Microwave-assisted extraction(MAE)
1.6.4 Enzyme assisted extraction(EAE)
1.6.5 Aqueous two-phase system
1.6.5.1 Alcohol/salt ATPS
1.7 Traditional deproteinization methods of polysaccharides
1.7.1 Sevag Method
1.7.2 Trichloroacetic acid method(TCA)
1.7.3 NaCl method
1.7.4 Calcium chloride method
1.8 Traditional purification methods
1.8.1 Column Chromatography
1.8.2 High-Pressure Liquid Chromatography(HPLC)
1.8.3 Gas Chromatography
1.8.4 Ethanol graded precipitation
1.9 Significance and Justification of the study
1.10 Specific objectives
CHAPTER 2 DUAL-FREQUENCY ULTRASOUND-ASSISTED ALCOHOL/SALT AQUEOUS TWO-PHASE EXTRACTION AND PURIFICATION OF ASTRAGALUS POLYSACCHARIDES
2.1 Introduction
2.2 MATERIALS AND METHODS
2.2.1 Materials and chemicals
2.2.2 Plant material preparation
2.2.3 Pre-treatment of Astragalus root powder
2.2.4 Phase diagram
2.2.5 Selection of ethanol/salt ATPS
2.2.6 Dual-frequency ultrasound-assisted aqueous two-phase extraction(DFu-AATPE) ..
2.2.7 Conventinal extraction experiments
2.2.8 Response surface methodology
2.2.9 Desalination
2.2.10 Thermodynamics
2.2.11 Chemical analysis
2.2.12 Structural Analysis
2.2.12.1 Monosaccharide Composition
2.2.12.2 Molecular Weight
2.2.12.3 Morphological structure of polysaccharides
2.2.13 Spectral analysis
2.2.13.1 Fourier Transform Infrared Spectroscopy
2.2.13.2 Ultra-Violet Scanning
2.2.14 Radical scavenging activity assays
2.2.14.1 Hydroxyl radical scavenging activity
2.2.14.2 DPPH radical scavenging activity
2.2.15 Statistical analysis
2.3 RESULTS AND DISCUSSION
2.3.1 Phase diagram
2.3.2 Selection of ATPS
2.3.2.1 Effect of salt concentration on the partition coefficient of APS in ATPS
2.3.2.2 Effect of ethanol concentration on the partition coefficient of APS in ATPS
2.3.2.3 Effect of temperature on the partition coefficient of APS in ATPS
2.3.3 Single-factor experiments on DFu-AATPE
2.3.3.1 Effect of temperature on APS yield
2.3.3.2 Effect of dual-frequency ultrasound on APS yield
2.3.3.3 Effect of(NH4)2SO4 concentration on APS yield
2.3.3.4 Effect of Ethanol addition on APS yield
2.3.3.5 Effect of ultrasound power on APS yield
2.3.3.6 Effect of extraction time on APS yield
2.3.4 Optimization
2.3.4.1 Model fitting
2.3.4.2 Response Surface Analysis
2.3.4.3 Validation of the model
2.3.5 Primary Characterization of APS polysaccharides
2.3.5.1 Carbohydrate content
2.3.5.2 Residual rate of protein after extraction and purification
2.3.6 Thermodynamics
2.3.7 Desalination
2.3.8 Structural composition
2.3.8.1 Monosaccharide Composition
2.3.8.2 Molecular weight determination(HPGPC)
2.3.8.3 Scanning Electron Microscopy
2.3.9 Spectral analysis
2.3.9.1 Fourier Transform-Infrared Spectroscopy
2.3.9.2 UV-Vis analysis
2.3.10 Anti-radical activities
2.3.10.1 Hydroxyl scavenging activity
2.3.10.2 DPPH scavenging assay
2.3.11 Mechanism of DFu-AATPE of Astragalus polysaccharides
2.3.12 Comparison with other extraction techniques
2.4 Conclusion
CHAPTER 3 SYNERGIZED SUBCRITICAL-ULTRASOUND-ASSISTED AQUEOUS TWO-PHASE EXTRACTION,PURIFICATION,AND CHARACTERIZATION OF LENTINUS EDODES POLYSACCHARIDES
3.1 Introduction
3.2 MATERIALS AND METHODS
3.2.1 Chemicals and reagents
3.2.2 Plant material and pretreatment
3.2.3 Subcritical water extraction
3.2.4 Purification
3.2.5 Comparison of Extraction methods
3.2.6 Response Surface Methodology
3.2.7 Ultrasound-assisted desalination
3.2.8 Chemical Composition of LEPs
3.2.9 Structural characterization
3.2.9.1 Monosaccharide Composition
3.2.9.2 Molecular Weight determination
3.2.9.3 Congo red
3.2.9.4 Scanning Electron Microscopy
3.2.10 Spectral analysis
3.2.10.1 Fourier Transform IR spectrophotometer(FT-IR) analysis
3.2.10.2 UV-vis analysis
3.2.11 Radical scavenging activity assays
3.2.11.1 2.2-diphenyl;-1 picrylhydrazyl(DPPH) scavenging activity
3.2.11.2 Hydroxyl radical scavenging activity
3.2.11.3 2 2-azino-bis(3-ethylbenzothiazoline-6 sulfonic acid) ABTS Scavenging activity
3.2.12 Statistical analysis
3.3 RESULTS AND DISCUSSION
3.3.1 Yield of Lentinus edodes polysaccharides
3.3.2 Optimization
3.3.2.1 Model fitting
3.3.2.2 Effect of independent variables on the LEP yield
3.3.2.3 Model Verification
3.3.3 Mechanism of Multi-frequency ultrasound-assisted aqueous two-phase extraction(MFU-AATPE)of Lentinus edodes polysaccharides
3.3.4 Chemical composition
3.3.5 Desalination
3.3.6 Structural Analysis
3.3.6.1 Monosaccharide Composition
3.3.6.2 Molecular weight
3.3.6.3 Congo red
3.3.6.4 Scanning Electron Microscopy(SEM)
3.3.7 Spectral analysis
3.3.7.1.Fourier Transform Infrared(FTIR)
3.3.7.2.Ultraviolet-visible spectroscopy analysis
3.3.8 Radical Scavenging Activity Assays
3.3.8.1 2.2-diphenyl;-1 picrylhydrazyl(DPPH)scavenging activity
3.3.8.2 Hydroxyl scavenging activity
3.3.8.3 2,2-azino-bis(3-ethylbenzothiazoline-6 sulfonic acid)ABTS scavenging activity
3.3.9 Comparison with other extraction techniques
3.5 Conclusion
CHAPTER 4 CONCLUSION AND FUTURE WORK
4.1 General Conclusion
4.2 Novelty
4.3 Recommendations
REFERENCES
APPENDIX
【参考文献】：
期刊论文
[1]交替双频逆流超声辅助提取条斑紫菜蛋白和多糖[J]. 曲文娟,马海乐,王婷,郑惠华. 农业工程学报. 2013(01)
[2]日粮添加黄芪多糖对蛋鸡机体抗氧化能力和鸡蛋品质的影响[J]. 左兆云,杨维仁,杨在宾,姜淑贞,张桂国,赵旭. 中国兽医学报. 2012(01)
[3]超声联合酶法提取黄芪总多糖的影响因素分析[J]. 贲永光,吴铮超. 广东药学院学报. 2010(02)
[4]膜荚黄芪与蒙古黄芪植物学特征分析[J]. 燕玲,宛涛,张众,汪西荣,孙文斌. 内蒙古农业大学学报(自然科学版). 2001(04)
本文编号：3383339
【文章来源】：江苏大学江苏省
【文章页数】：124 页
【学位级别】：硕士
【文章目录】：
ACKNOWLEDGEMENT
ABSTRACT
摘要
ABBREVIATIONS
CHAPTER 1 INTRODUCTION AND RESEARCH OBJECTIVES
1.1 General background
1.2 Astragalus membranaceus
1.2.1 Composition
1.2.2 Health-related properties
1.3 Lentinus edodes
1.3.1 Composition
1.3.2 Health-related properties
1.4 Extraction of polysaccharides
1.5 Conventional polysaccharide extraction methods
1.5.1 Hot water extraction
1.5.2 Reflux extraction
1.5.3 Complex enzyme hydrolysis method
1.6 Novel polysaccharide extraction methods
1.6.1 Ultrasound-assisted extraction
1.6.1.1 Multi-frequency ultrasound
1.6.2 Subcritical water extraction(SWE)
1.6.3 Microwave-assisted extraction(MAE)
1.6.4 Enzyme assisted extraction(EAE)
1.6.5 Aqueous two-phase system
1.6.5.1 Alcohol/salt ATPS
1.7 Traditional deproteinization methods of polysaccharides
1.7.1 Sevag Method
1.7.2 Trichloroacetic acid method(TCA)
1.7.3 NaCl method
1.7.4 Calcium chloride method
1.8 Traditional purification methods
1.8.1 Column Chromatography
1.8.2 High-Pressure Liquid Chromatography(HPLC)
1.8.3 Gas Chromatography
1.8.4 Ethanol graded precipitation
1.9 Significance and Justification of the study
1.10 Specific objectives
CHAPTER 2 DUAL-FREQUENCY ULTRASOUND-ASSISTED ALCOHOL/SALT AQUEOUS TWO-PHASE EXTRACTION AND PURIFICATION OF ASTRAGALUS POLYSACCHARIDES
2.1 Introduction
2.2 MATERIALS AND METHODS
2.2.1 Materials and chemicals
2.2.2 Plant material preparation
2.2.3 Pre-treatment of Astragalus root powder
2.2.4 Phase diagram
2.2.5 Selection of ethanol/salt ATPS
2.2.6 Dual-frequency ultrasound-assisted aqueous two-phase extraction(DFu-AATPE) ..
2.2.7 Conventinal extraction experiments
2.2.8 Response surface methodology
2.2.9 Desalination
2.2.10 Thermodynamics
2.2.11 Chemical analysis
2.2.12 Structural Analysis
2.2.12.1 Monosaccharide Composition
2.2.12.2 Molecular Weight
2.2.12.3 Morphological structure of polysaccharides
2.2.13 Spectral analysis
2.2.13.1 Fourier Transform Infrared Spectroscopy
2.2.13.2 Ultra-Violet Scanning
2.2.14 Radical scavenging activity assays
2.2.14.1 Hydroxyl radical scavenging activity
2.2.14.2 DPPH radical scavenging activity
2.2.15 Statistical analysis
2.3 RESULTS AND DISCUSSION
2.3.1 Phase diagram
2.3.2 Selection of ATPS
2.3.2.1 Effect of salt concentration on the partition coefficient of APS in ATPS
2.3.2.2 Effect of ethanol concentration on the partition coefficient of APS in ATPS
2.3.2.3 Effect of temperature on the partition coefficient of APS in ATPS
2.3.3 Single-factor experiments on DFu-AATPE
2.3.3.1 Effect of temperature on APS yield
2.3.3.2 Effect of dual-frequency ultrasound on APS yield
2.3.3.3 Effect of(NH4)2SO4 concentration on APS yield
2.3.3.4 Effect of Ethanol addition on APS yield
2.3.3.5 Effect of ultrasound power on APS yield
2.3.3.6 Effect of extraction time on APS yield
2.3.4 Optimization
2.3.4.1 Model fitting
2.3.4.2 Response Surface Analysis
2.3.4.3 Validation of the model
2.3.5 Primary Characterization of APS polysaccharides
2.3.5.1 Carbohydrate content
2.3.5.2 Residual rate of protein after extraction and purification
2.3.6 Thermodynamics
2.3.7 Desalination
2.3.8 Structural composition
2.3.8.1 Monosaccharide Composition
2.3.8.2 Molecular weight determination(HPGPC)
2.3.8.3 Scanning Electron Microscopy
2.3.9 Spectral analysis
2.3.9.1 Fourier Transform-Infrared Spectroscopy
2.3.9.2 UV-Vis analysis
2.3.10 Anti-radical activities
2.3.10.1 Hydroxyl scavenging activity
2.3.10.2 DPPH scavenging assay
2.3.11 Mechanism of DFu-AATPE of Astragalus polysaccharides
2.3.12 Comparison with other extraction techniques
2.4 Conclusion
CHAPTER 3 SYNERGIZED SUBCRITICAL-ULTRASOUND-ASSISTED AQUEOUS TWO-PHASE EXTRACTION,PURIFICATION,AND CHARACTERIZATION OF LENTINUS EDODES POLYSACCHARIDES
3.1 Introduction
3.2 MATERIALS AND METHODS
3.2.1 Chemicals and reagents
3.2.2 Plant material and pretreatment
3.2.3 Subcritical water extraction
3.2.4 Purification
3.2.5 Comparison of Extraction methods
3.2.6 Response Surface Methodology
3.2.7 Ultrasound-assisted desalination
3.2.8 Chemical Composition of LEPs
3.2.9 Structural characterization
3.2.9.1 Monosaccharide Composition
3.2.9.2 Molecular Weight determination
3.2.9.3 Congo red
3.2.9.4 Scanning Electron Microscopy
3.2.10 Spectral analysis
3.2.10.1 Fourier Transform IR spectrophotometer(FT-IR) analysis
3.2.10.2 UV-vis analysis
3.2.11 Radical scavenging activity assays
3.2.11.1 2.2-diphenyl;-1 picrylhydrazyl(DPPH) scavenging activity
3.2.11.2 Hydroxyl radical scavenging activity
3.2.11.3 2 2-azino-bis(3-ethylbenzothiazoline-6 sulfonic acid) ABTS Scavenging activity
3.2.12 Statistical analysis
3.3 RESULTS AND DISCUSSION
3.3.1 Yield of Lentinus edodes polysaccharides
3.3.2 Optimization
3.3.2.1 Model fitting
3.3.2.2 Effect of independent variables on the LEP yield
3.3.2.3 Model Verification
3.3.3 Mechanism of Multi-frequency ultrasound-assisted aqueous two-phase extraction(MFU-AATPE)of Lentinus edodes polysaccharides
3.3.4 Chemical composition
3.3.5 Desalination
3.3.6 Structural Analysis
3.3.6.1 Monosaccharide Composition
3.3.6.2 Molecular weight
3.3.6.3 Congo red
3.3.6.4 Scanning Electron Microscopy(SEM)
3.3.7 Spectral analysis
3.3.7.1.Fourier Transform Infrared(FTIR)
3.3.7.2.Ultraviolet-visible spectroscopy analysis
3.3.8 Radical Scavenging Activity Assays
3.3.8.1 2.2-diphenyl;-1 picrylhydrazyl(DPPH)scavenging activity
3.3.8.2 Hydroxyl scavenging activity
3.3.8.3 2,2-azino-bis(3-ethylbenzothiazoline-6 sulfonic acid)ABTS scavenging activity
3.3.9 Comparison with other extraction techniques
3.5 Conclusion
CHAPTER 4 CONCLUSION AND FUTURE WORK
4.1 General Conclusion
4.2 Novelty
4.3 Recommendations
REFERENCES
APPENDIX
【参考文献】：
期刊论文
[1]交替双频逆流超声辅助提取条斑紫菜蛋白和多糖[J]. 曲文娟,马海乐,王婷,郑惠华. 农业工程学报. 2013(01)
[2]日粮添加黄芪多糖对蛋鸡机体抗氧化能力和鸡蛋品质的影响[J]. 左兆云,杨维仁,杨在宾,姜淑贞,张桂国,赵旭. 中国兽医学报. 2012(01)
[3]超声联合酶法提取黄芪总多糖的影响因素分析[J]. 贲永光,吴铮超. 广东药学院学报. 2010(02)
[4]膜荚黄芪与蒙古黄芪植物学特征分析[J]. 燕玲,宛涛,张众,汪西荣,孙文斌. 内蒙古农业大学学报(自然科学版). 2001(04)
本文编号：3383339
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