亚硝基血红蛋白制备及在肉制品中的应用
发布时间：2021-10-30 11:57
本研究旨在开发具有良好着色稳定性、天然安全性、抗氧化性和抗菌性等特性的亚硝基血红蛋白（NO-Hb）作为色素添加剂,用以替代肉制品中亚硝酸盐。这种着色剂不仅可以使肉制品呈现出可观的颜色,而且还可以维持肉制品的颜色稳定性,使肉制品具有缓慢的褪色速度,合适的显色效果和良好的平衡性。这明显增强肉制品的颜色,减少残留的亚硝酸盐量,延长肉制品的保质期。实验研究集中于制备新型着色剂,即NO-Hb。通过使用副产品（牛血液）制备的NO-Hb具有亚硝酸盐替代品的应用前景,可提高颜色稳定性,并抑制病原菌。用于NO-Hb 提取的优化条件分别为 pH 6.0、VC 6.0 g/L、NaNO2 3.5 g/L 和转速 8000 r/min。含亚硝基血红蛋白的碎牛肉的IC50显着高于对照组（P<0.05）,IC50是含亚硝基血红蛋白的碎牛肉抑制50%DPPH阴离子时的浓度,含亚硝基血红蛋白的碎牛肉的IC50为10.53 mg/mL,而维生素C的IC50为9.84 mg/mL。含亚硝基血红蛋白和维生素C的碎牛肉的ABTS-SA显著高于对照组（P<0.05）。在4℃冰箱中冷藏1周后,含亚硝基血红蛋白的碎牛肉...
【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校
【文章页数】：255 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
ABBREVIATIONS
Chapter Ⅰ:Introduction and Literature Review
1 Research background
2 Hemoglobin chemical composition
2.1 Heme pigment found in myoglobin
2.2 Myoglobin and oxidative processes in biological systems
2.3 Hemoglobin profiles(source,structure,and properties)
2.4 Structural biochemistry/hemoglobin
3 Progress and functional of NO-Hb preparation
3.1 Meat color and meat product colorants status and development trend
3.2 Meat color
3.3 Meat product colorants status and development
3.4 Meat color measurement
4 Summary research of previous studied of NO-Hb approaches in the food industry
4.1 Summary of research evaluating(NO-Hb preparation)and used in meat colorant development
4.2 Summary of research evaluating used blood as food additives
4.3 Summary of research assessing current affecting in meat product's color
4.4 Summary of research evaluating changes in color and Mb effect atmosphere packaging
4.5 Summary of research evaluating antimicrobial significantly affecting meat color products
4.6 Summary of research evaluating processed and colorectal cancer
5 Common techniques in studying NO-Hb and minced beef chemical,physical and structural changes
5.1 Optimization process and preparation of NO-Hb from beef blood
5.2 Antioxidant activities of NO-Hb
5.3 Low-field nuclear magnetic resonance(LF-NMR)
5.4 Investigation of amino acid by HPLC technique
5.5 The influence of calcium with NO-Hb on the color stability in cooked beef
5.6 The effect of vitamin C on storage cooked beef mixed with NO-Hb and calcium
5.7 Investigation of minced beef volatiles by head space-solid phase microextraction technique(HS-SPME)/Gas Chromatography Mass Spectrometer(GC-MS)
5.8 Scanning electron microscopy(SEM)as a well-organized way for the minced beef study
5.9 Fluorescent microscope(FM)as an efficient way for minced beef microstructure study
5.10 Microstructure assay by Reflection confocal scanning laser microscopy(r CLSM)
5.11 Coupled techniques in studding meats chemical and physical evaluate
6 Dissertation aim of work
Chapter Ⅱ:Optimization process and preparation of Nitroso-hemoglobin(NO-Hb)
1 Materials and methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Chemicals
1.1.3 Instruments
1.2 Research methods
1.2.1 Preparation of NO-Hb
1.2.2 Single-factor and orthogonal experimental design
1.3 Statistical analysis
2 Results and Discussion
2.1 The optimization of p H on the preparation of NO-Hb
2.2 Development of an Enhancement Acquires Stabilizer for Na NO2
2.3 Optimizing of VC
2.4 Speed optimized by using single factor's experiments and orthogonal test
2.5 Time stabilizer for hemoglobin extractions by using UV test
2.6 Development of an Effective Compound Stabilizer for preparation of NO-Hb by an Orthogonal Test
2.6.1 Single factors and levels of the L9(34)orthogonal test
2.6.2 Evaluation of universal design of the orthogonal test compound stabilizers investigated protective the effect
2.6.3 Operating Conditions of Variance analysis significance of the orthogonal test
2.7 Verification tests
2.8 Antioxidant activity via the DPPH· -SA assay
2.9 Antioxidant capacity via the ABTS· +assay
2.10 Antioxidant activity and capacity of NO-Hb
3 Conclusion
Chapter Ⅲ:Nitroso-hemoglobin increased the color stability and inhibited the pathogenic bacteria in a minced beef model
1 Materials and methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Bacteria species
1.1.3 Chemicals
1.1.4 Instruments
1.1.5 Meat samples treating
1.2 Research methods
1.2.1 Microbiological analysis
1.2.2 Color measurements
1.2.3 Low-Field Nuclear Magnetic Resonance(LF-NMR)
1.3 Statistical analysis
2 Results and discussion
2.1 The effect of NO-Hb on individual pathogenic microbial counts(log10 CFU/g) (1:1)injected in a fresh beef
2.2 The effect of NO-Hb on combined bacteria counts during7-10 d of storage
2.3 The influence of LAB on combined pathogenic bacteria inoculated in different ratios after7 d of storage
2.4 The effect of NO-Hb on combined microbial during13-16 d of storage
2.5 Color formation in the minced beef
2.6 Relaxation time analysis of raw minced beef
2.7 Effect of bacteria and storage period on relaxation time T2
3 Conclusion
Chapter Ⅳ:Nitroso-hemoglobin-ginger conjugates inhibit the bacterial growth and increase the color stability in a mined beef model:A study of anti-pathogenic effects on water-binding
1 Materials and Methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Preparation of ginger extracts
1.1.3 Bacteria species
1.1.4 Chemicals
1.1.5 Instruments
1.1.6 Meat samples treating
1.2 Analytical methods
1.2.1 Microbiological analysis
1.2.2 Low-field nuclear magnetic resonance(LF-NMR)
1.3 Statistical analysis
2 Results and discussion
2.1 Effects of NO-Hb with VC,Ca,and ginger on the bacterial growth in a minced beef model
2.2 Color formation in the minced beef models
2.3 Relaxation time analysis of raw minced beef with different ingredients and pathogens
2.4 Effect of bacteria and storage period on relaxation time T2
3 Conclusion
Chapter Ⅴ: The effect of nitroso-hemoglobin-arabic gum on the bacterial counts and volatile compounds in minced beef at different storage conditions: A combined LF-NMR, HPLC and HS-SPME/GC-MS study
1 Materials and Methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Bacteria species
1.1.3 Chemicals
1.1.4 Instruments
1.1.5 Meat samples treating
1.2 Research methods
1.2.1 Microbiological analysis
1.2.2 Proximate analysis
1.2.3 HS-SPME-GC-Ms
1.3 Statistical analysis
2 Results and discussion
2.1 Total bacterial count in fresh meat
2.2 LF-NMR results
2.2.1 Three relaxation times(T2)and their corresponding water population percentages(A2)
2.2.2 Effect of NO-Hb+ Gu A and storage on peak area properties
2.3 Amino acids(AAs)composition of mined beef
2.4 Qualitative analysis:identification of volatile compounds in raw minced beef
2.5 Characterization of volatile groups by multivariate analyses
2.5.1 Volatiles principal component analysis(PCA)
3 Conclusion
Chapter Ⅵ:The effects of Nitroso-hemoglobin on minced beef microstructure pattern during the storage
1 Materials and methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Chemicals
1.1.3 Instruments
1.1.4 Sample preparation
1.2 Analytical methods
1.2.1 Microstructure assay by Scanning Electron Microscopy(SEM)
1.2.2 Microstructure assay by Fluorescent microscope(FM)
1.2.3 Reflection confocal scanning laser microscopy(r CLSM)
2 Results and discussion
1.1 Microstructure evaluate by Scanning electron microscope(SEM)
2.1.1 Scanning Electron Microscope
2.1.2 Scanning Electron Microscope
1.2 Microstructure assay by Fluorescence microscope(FM)
2.2.1 Nile blue Fluorescence micrographs
2.2.2 Nile red Fluorescence micrographs
1.3.3 Microstructure assay by Reflection confocal scanning laser microscopy(rCLSM)
3 Conclusion
Chapter Ⅶ:Synergistic effect of NO-Hb with calcium and vitamin C on the color stability of cooked minced beef during storage
1 Materials and Methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Chemicals
1.1.3 Instruments
1.2 Research methods
1.2.1 Samples Preparation
1.2.2 Color determination
1.2.3 UV-VIS Spectrophotometer
1.2.4 Low-field nuclear magnetic resonance(LF-NMR)
1.3 Statistical Analysis
2 Results and dissections
2.1 Color Change
2.2 Improvement of color stability
2.3 The effects of thermal on minced beef color stability
2.4 The effects of the refrigerator on Color parameters of fresh beef
2.5 The effects of frozen on Color parameters of fresh frozen beef
2.6 The effect of frozen on cooking beef with Ca during storage(up to28 days)
2.7 The effect of adding VC on cooking beef with Ca during frozen storage(up to28 days)
2.8 Relaxation time analysis of raw minced beef and cooked minced beef with/without nitrite and NO-Hb
2.9 Relaxation time analysis of raw feef
2.10 Relaxation time analysis of cooked minced beef with nitrite and NO-Hb
3 Conclusion
Chapter Ⅷ:Conclusion and perspective
1 Main conclusion
2 Innovation points
3 Future Works
References
Supplementary materials
Appendix A.Flowchart of preparing Nitroso-hemoglobin.It is made from fresh blood(red cells)by the use of by-products
Appendix B.The effect of nitroso-hemoglobin-arabic gum+Ca+ VC on the bacterial counts cook and storage of minced beef at different storage conditions:A combined low-field NMR,HPLC,and HS-SPME/GC-MS study
Appendix C.Supplementary Figures:The effect of NO-Hb on bacteria counts such as(Total bacterial counts(TBC),E.coli,Staphylococcus,Pseudomonas,salmonella,and lactic acid bacteria(LAB)during two weeks of storage
Appendix D.Peak,retention time,peak area,and concentration(%)of HPLC figures
Appendix E.The influence of calcium and VC with NO-Hb on the color stability in cooked and storage minced beef
Appendix F.Recent published articles from the Ph.D.dissertation:
Appendix G.International Conference
Appendix H.Acknowledgement
Appendix I.Dedication
【参考文献】：
期刊论文
[1]Animals as sources of food-borne pathogens: A review[J]. Norma Heredia,Santos García. Animal Nutrition. 2018(03)
[2]麦角硫因处理对采后双孢蘑菇品质的影响[J]. 靳祯亮,黄琦辉,周巧丽,姜天甲,郑小林. 中国食品学报. 2017(03)
[3]蛋清溶菌酶的分离纯化及其抑菌活性研究[J]. 张黎丽,马梦琳,李晨熙,常允康. 生物化工. 2017(01)
[4]新型腌制色素糖基化亚硝基血红蛋白的研究进展[J]. 曹磊,宋玉,汪晓鸣,程仁功,陈海峰,刘超. 安徽农业科学. 2013(07)
[5]肉制品中亚硝酸盐替代物的研究进展及应用[J]. 张红涛,孔保华,蒋亚男. 包装与食品机械. 2012(03)
本文编号：3466689
【文章来源】：华中农业大学湖北省 211工程院校 教育部直属院校
【文章页数】：255 页
【学位级别】：博士
【文章目录】：
摘要
ABSTRACT
ABBREVIATIONS
Chapter Ⅰ:Introduction and Literature Review
1 Research background
2 Hemoglobin chemical composition
2.1 Heme pigment found in myoglobin
2.2 Myoglobin and oxidative processes in biological systems
2.3 Hemoglobin profiles(source,structure,and properties)
2.4 Structural biochemistry/hemoglobin
3 Progress and functional of NO-Hb preparation
3.1 Meat color and meat product colorants status and development trend
3.2 Meat color
3.3 Meat product colorants status and development
3.4 Meat color measurement
4 Summary research of previous studied of NO-Hb approaches in the food industry
4.1 Summary of research evaluating(NO-Hb preparation)and used in meat colorant development
4.2 Summary of research evaluating used blood as food additives
4.3 Summary of research assessing current affecting in meat product's color
4.4 Summary of research evaluating changes in color and Mb effect atmosphere packaging
4.5 Summary of research evaluating antimicrobial significantly affecting meat color products
4.6 Summary of research evaluating processed and colorectal cancer
5 Common techniques in studying NO-Hb and minced beef chemical,physical and structural changes
5.1 Optimization process and preparation of NO-Hb from beef blood
5.2 Antioxidant activities of NO-Hb
5.3 Low-field nuclear magnetic resonance(LF-NMR)
5.4 Investigation of amino acid by HPLC technique
5.5 The influence of calcium with NO-Hb on the color stability in cooked beef
5.6 The effect of vitamin C on storage cooked beef mixed with NO-Hb and calcium
5.7 Investigation of minced beef volatiles by head space-solid phase microextraction technique(HS-SPME)/Gas Chromatography Mass Spectrometer(GC-MS)
5.8 Scanning electron microscopy(SEM)as a well-organized way for the minced beef study
5.9 Fluorescent microscope(FM)as an efficient way for minced beef microstructure study
5.10 Microstructure assay by Reflection confocal scanning laser microscopy(r CLSM)
5.11 Coupled techniques in studding meats chemical and physical evaluate
6 Dissertation aim of work
Chapter Ⅱ:Optimization process and preparation of Nitroso-hemoglobin(NO-Hb)
1 Materials and methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Chemicals
1.1.3 Instruments
1.2 Research methods
1.2.1 Preparation of NO-Hb
1.2.2 Single-factor and orthogonal experimental design
1.3 Statistical analysis
2 Results and Discussion
2.1 The optimization of p H on the preparation of NO-Hb
2.2 Development of an Enhancement Acquires Stabilizer for Na NO2
2.3 Optimizing of VC
2.4 Speed optimized by using single factor's experiments and orthogonal test
2.5 Time stabilizer for hemoglobin extractions by using UV test
2.6 Development of an Effective Compound Stabilizer for preparation of NO-Hb by an Orthogonal Test
2.6.1 Single factors and levels of the L9(34)orthogonal test
2.6.2 Evaluation of universal design of the orthogonal test compound stabilizers investigated protective the effect
2.6.3 Operating Conditions of Variance analysis significance of the orthogonal test
2.7 Verification tests
2.8 Antioxidant activity via the DPPH· -SA assay
2.9 Antioxidant capacity via the ABTS· +assay
2.10 Antioxidant activity and capacity of NO-Hb
3 Conclusion
Chapter Ⅲ:Nitroso-hemoglobin increased the color stability and inhibited the pathogenic bacteria in a minced beef model
1 Materials and methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Bacteria species
1.1.3 Chemicals
1.1.4 Instruments
1.1.5 Meat samples treating
1.2 Research methods
1.2.1 Microbiological analysis
1.2.2 Color measurements
1.2.3 Low-Field Nuclear Magnetic Resonance(LF-NMR)
1.3 Statistical analysis
2 Results and discussion
2.1 The effect of NO-Hb on individual pathogenic microbial counts(log10 CFU/g) (1:1)injected in a fresh beef
2.2 The effect of NO-Hb on combined bacteria counts during7-10 d of storage
2.3 The influence of LAB on combined pathogenic bacteria inoculated in different ratios after7 d of storage
2.4 The effect of NO-Hb on combined microbial during13-16 d of storage
2.5 Color formation in the minced beef
2.6 Relaxation time analysis of raw minced beef
2.7 Effect of bacteria and storage period on relaxation time T2
3 Conclusion
Chapter Ⅳ:Nitroso-hemoglobin-ginger conjugates inhibit the bacterial growth and increase the color stability in a mined beef model:A study of anti-pathogenic effects on water-binding
1 Materials and Methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Preparation of ginger extracts
1.1.3 Bacteria species
1.1.4 Chemicals
1.1.5 Instruments
1.1.6 Meat samples treating
1.2 Analytical methods
1.2.1 Microbiological analysis
1.2.2 Low-field nuclear magnetic resonance(LF-NMR)
1.3 Statistical analysis
2 Results and discussion
2.1 Effects of NO-Hb with VC,Ca,and ginger on the bacterial growth in a minced beef model
2.2 Color formation in the minced beef models
2.3 Relaxation time analysis of raw minced beef with different ingredients and pathogens
2.4 Effect of bacteria and storage period on relaxation time T2
3 Conclusion
Chapter Ⅴ: The effect of nitroso-hemoglobin-arabic gum on the bacterial counts and volatile compounds in minced beef at different storage conditions: A combined LF-NMR, HPLC and HS-SPME/GC-MS study
1 Materials and Methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Bacteria species
1.1.3 Chemicals
1.1.4 Instruments
1.1.5 Meat samples treating
1.2 Research methods
1.2.1 Microbiological analysis
1.2.2 Proximate analysis
1.2.3 HS-SPME-GC-Ms
1.3 Statistical analysis
2 Results and discussion
2.1 Total bacterial count in fresh meat
2.2 LF-NMR results
2.2.1 Three relaxation times(T2)and their corresponding water population percentages(A2)
2.2.2 Effect of NO-Hb+ Gu A and storage on peak area properties
2.3 Amino acids(AAs)composition of mined beef
2.4 Qualitative analysis:identification of volatile compounds in raw minced beef
2.5 Characterization of volatile groups by multivariate analyses
2.5.1 Volatiles principal component analysis(PCA)
3 Conclusion
Chapter Ⅵ:The effects of Nitroso-hemoglobin on minced beef microstructure pattern during the storage
1 Materials and methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Chemicals
1.1.3 Instruments
1.1.4 Sample preparation
1.2 Analytical methods
1.2.1 Microstructure assay by Scanning Electron Microscopy(SEM)
1.2.2 Microstructure assay by Fluorescent microscope(FM)
1.2.3 Reflection confocal scanning laser microscopy(r CLSM)
2 Results and discussion
1.1 Microstructure evaluate by Scanning electron microscope(SEM)
2.1.1 Scanning Electron Microscope
2.1.2 Scanning Electron Microscope
1.2 Microstructure assay by Fluorescence microscope(FM)
2.2.1 Nile blue Fluorescence micrographs
2.2.2 Nile red Fluorescence micrographs
1.3.3 Microstructure assay by Reflection confocal scanning laser microscopy(rCLSM)
3 Conclusion
Chapter Ⅶ:Synergistic effect of NO-Hb with calcium and vitamin C on the color stability of cooked minced beef during storage
1 Materials and Methods
1.1 Materials
1.1.1 Raw materials
1.1.2 Chemicals
1.1.3 Instruments
1.2 Research methods
1.2.1 Samples Preparation
1.2.2 Color determination
1.2.3 UV-VIS Spectrophotometer
1.2.4 Low-field nuclear magnetic resonance(LF-NMR)
1.3 Statistical Analysis
2 Results and dissections
2.1 Color Change
2.2 Improvement of color stability
2.3 The effects of thermal on minced beef color stability
2.4 The effects of the refrigerator on Color parameters of fresh beef
2.5 The effects of frozen on Color parameters of fresh frozen beef
2.6 The effect of frozen on cooking beef with Ca during storage(up to28 days)
2.7 The effect of adding VC on cooking beef with Ca during frozen storage(up to28 days)
2.8 Relaxation time analysis of raw minced beef and cooked minced beef with/without nitrite and NO-Hb
2.9 Relaxation time analysis of raw feef
2.10 Relaxation time analysis of cooked minced beef with nitrite and NO-Hb
3 Conclusion
Chapter Ⅷ:Conclusion and perspective
1 Main conclusion
2 Innovation points
3 Future Works
References
Supplementary materials
Appendix A.Flowchart of preparing Nitroso-hemoglobin.It is made from fresh blood(red cells)by the use of by-products
Appendix B.The effect of nitroso-hemoglobin-arabic gum+Ca+ VC on the bacterial counts cook and storage of minced beef at different storage conditions:A combined low-field NMR,HPLC,and HS-SPME/GC-MS study
Appendix C.Supplementary Figures:The effect of NO-Hb on bacteria counts such as(Total bacterial counts(TBC),E.coli,Staphylococcus,Pseudomonas,salmonella,and lactic acid bacteria(LAB)during two weeks of storage
Appendix D.Peak,retention time,peak area,and concentration(%)of HPLC figures
Appendix E.The influence of calcium and VC with NO-Hb on the color stability in cooked and storage minced beef
Appendix F.Recent published articles from the Ph.D.dissertation:
Appendix G.International Conference
Appendix H.Acknowledgement
Appendix I.Dedication
【参考文献】：
期刊论文
[1]Animals as sources of food-borne pathogens: A review[J]. Norma Heredia,Santos García. Animal Nutrition. 2018(03)
[2]麦角硫因处理对采后双孢蘑菇品质的影响[J]. 靳祯亮,黄琦辉,周巧丽,姜天甲,郑小林. 中国食品学报. 2017(03)
[3]蛋清溶菌酶的分离纯化及其抑菌活性研究[J]. 张黎丽,马梦琳,李晨熙,常允康. 生物化工. 2017(01)
[4]新型腌制色素糖基化亚硝基血红蛋白的研究进展[J]. 曹磊,宋玉,汪晓鸣,程仁功,陈海峰,刘超. 安徽农业科学. 2013(07)
[5]肉制品中亚硝酸盐替代物的研究进展及应用[J]. 张红涛,孔保华,蒋亚男. 包装与食品机械. 2012(03)
本文编号：3466689
本文链接：https://www.wllwen.com/projectlw/qgylw/3466689.html
