Introduction
Based on his experience in teaching "Food Analysis", "Industrial Fermentation Analysis" and other courses for more than 20 years, and in the past 10 years, he has taught "Bioengineering" to undergraduates in bioengineering. The self-edited lecture content of the "Analysis" course is compiled with reference to other professional analysis textbooks published in the past three years.
The book consists of 18 chapters, including introduction, basic knowledge of bioengineering analysis, physical analysis, chemical analysis, ultraviolet-visible absorption spectroscopy, fluorescence spectrophotometry, thin layer chromatography, gas chromatography , High-performance liquid chromatography, electrophoresis and high-performance capillary electrophoresis, nuclear magnetic resonance spectroscopy, biological mass spectrometry, bioassay, enzymatic analysis, immunoassay, biosensor, other new analytical and detection technologies and new sample preparation Technology.
This book is compiled based on the principles of commonly used analysis and detection methods in the field of bioengineering. The book consists of 18 chapters, including introduction, basic knowledge of bioengineering analysis, physical analysis, chemical analysis, ultraviolet and visible absorption spectroscopy, fluorescence spectrophotometry, thin layer chromatography, gas chromatography, high performance liquid chromatography, Electrophoresis and high-performance capillary electrophoresis, nuclear magnetic resonance spectroscopy, biological mass spectrometry, bioassay, enzymatic analysis, immunoassay, biosensor, other new analytical techniques and new techniques for sample preparation. This book can be used as a teaching or reference book for undergraduates and postgraduates in bioengineering, biochemical, biotechnology, food quality and safety and other majors in colleges and universities related to analysis and detection technology courses; it can also be used as a reference book for technical supervision, quality inspection, health and epidemic prevention. A reference book for quality inspection technicians in food and drug quality management departments, engineers and technicians in units engaged in chemical and chemical engineering, biopharmaceuticals, microbiological engineering, biochemical product production, and environmental protection and monitoring, as well as technical personnel in related scientific research institutions.
Contents
Chapter 1 Introduction 1
The meaning of bioengineering and its research field 1
Bioengineering and its subject foundation 1
The main research fields of bioengineering and their interrelationships 2
2 The fields of bioengineering services 5
1?2?1Agriculture, aquaculture and food Industry 5
1?2?2Human health business6
1?2?3Resources and energy8
1?2?4Environmental protection 9< /p>
1?3 Tasks, functions, contents and characteristics of bioengineering analysis 9
1?3?1 Tasks and functions of bioengineering analysis 9
1? 3?2 Contents and characteristics of bioengineering analysis 11
1?4 Book writing ideas and usage suggestions 13
1?4?1 Writing ideas 13
< p>Recommendations for use 13Chapter 2 Basic knowledge of bioengineering analysis 14
2?1 Overview 14
2?1?1 Position in bioengineering analysis 14
2?1?2 Purpose of sample pretreatment 14
2?1?3 Evaluation criteria for sample pretreatment 15
2?2 Collection, preparation and storage of analytical samples16
2?2?1Sample collection16
2?2?2Sample preparation19
2?2?3 Sample preservation 19
2?3 General method of biological sample pretreatment 21
2?3?1 dissolution method 21
2?3?2Distillation 22
2?3?3 Dry ashing 22
2?3?4 Wet digestion 22
2? 3? 5 Oxygen bottle combustion method 22
2? 3? 6 Ultrasonic method 23
2? 4 Special method for biological sample pretreatment 24
2? 4?1 Separation and purification of biologically active substances 24
2?4?2 Cell disruption and solid-liquid separation 27
2?4?3 Separation of inclusion bodies 29
2?4?2 p>
2?4?4 liquid-liquid extraction 31
2?4?5 supercritical extraction 33
2?4?6 precipitation 33
2?4?7 membrane separation 34
2?4?8 chromatographic separation technology 34
2?5 error and the choice of analysis method 48
2 ?5?1 Error 48
2?5?2 Choice of analysis method 49
Chapter 3 Physical Analysis 52
3?1 Overview 52< /p>
3?2 Common physical analysis method 52
3?2?1 Density and relative density method 52
3?2?2 Refraction method 56
3?2?3 Polarization 60
Chapter 4 Chemical Analysis Methods 67
4?1 Overview 67
4?2 Application examples of acid-base titration 67
4?2 ?1Determination of total acid, volatile acid and non-volatile acid in liquor Determination of the total esters in the 4?2?4 formaldehyde titration method to determine the total amino acid content 69
4?2?5 trace Kjeldahl (Kjeldahl) nitrogen determination method 69
p>4?2?6 Salicylic acid determination 71
4?3 redox titration application example 72
4?3?1 reducing sugar determination ——Direct titration 72
4?3?2 potassium permanganate titration method for the determination of calcium 74
4?3?3 determination of total aldehydes in liquor 75
4?3?4 Determination of tannins in hops 76
4?3?5 Quantitative determination of vitamin C——2,6? Dichloroindophenol titration 77
< p>4?4 Coordination titration method application example 804?4?1 Hardness determination of brewing water 80
4?4?2 Determination of calcium content in brewing water 81
Application example of the 4?5 gravimetric method 82
4?5?1 Determination of total ash 82
4?5?2 Phosphorus in biochemical raw materials Determination of 84
4?5?3 starch raw material moisture determination-direct drying method 85
4?5?4 crude fat content determination-Soxhlet extraction method 85
Chapter 5 Ultraviolet Visible Absorption Spectroscopy Analysis 87
5?1 Overview 87
5?2 Method Principle 87
5?2?1 Ultraviolet and visible absorption spectra of organic compounds 87
5?2?2 Ultraviolet? Visible absorption spectra of inorganic compounds 89
5?2?3 Lambertian? Beer's Law of Absorption 89
5?3 Instrument structure and principle 90
5?3?1 Radiation light source 90
5?3?2 Spectroscope 90< /p>
5?3?3 absorption pool 91
5?3?4 detector 91
5?3?5 recorder and signal display system 91
p>5?4 Experimental technology 91
5?4?1 sample preparation 91
5?4?2 determination conditions selection 91
5?4?3 reaction conditions selection 92
5?4?4 reference solution selection 92
5?4?5 coexisting ion interference elimination method 93
p>5?4?6 The exact calculation of apparent molar absorption coefficient 93
5?5 UV? Visible absorption spectrum application range 93
5?5?1 Qualitative analysis 93
5?5?2 purity identification 93
5?5?3 quantitative determination 94
5?6 UV? Visible absorption spectroscopy in bioengineering analysis Application of 95
5?6?1 Coomassie brilliant blue method to determine protein content 95
5?6?2RNA quantitative determination 96
5?6?3 Quantitative Determination of Serum Cholesterol 98
Chapter 6 Fluorescence Spectrophotometry 99
6?1 Overview 99
6?2 Method Principle 99
6?2?1 excitation and emission spectra 101
6?2?2 fluorescence quantum yield and molecular structure 103
6?3 fluorescence analysis 105
p>6?3?1 Basic principles and main instruments 105
6?3?2 Fluorescence spectrophotometer classification 106
6?3?3 affects fluorescence Factors of analysis 107
6?3?4 fluorescence analysis method 110
6?3?5F?4500 (HITACHI) fluorescence spectrophotometer introduction 110
Application of 6?4 Fluorescence Spectrophotometry in Bioengineering Analysis 114
6?4?1 Vitamin E Fluorescence Spectrophotometry 115
6?4?2 Serum Fluorescence spectrophotometric determination of chloroquine 116
6?4?3 Fluorescence spectrophotometric determination of ampicillin in urine 117
Chapter 7 Thin layer chromatography 118
7?1 Overview 118
7?2 Basic Principles of Thin Layer Chromatography 119
7?3 Operation Technology of Thin Layer Chromatography 121
7?3?1 Carrier plate and adsorbent selection 121
7?3?2 Preparation of thin layer plate 123
7?3?3 spot sample 124
< p>7?3?4 Expand 1267?3?5 Positioning and color rendering 127
7?3?6 Qualitative 127
7?4 Thin-layer quantitative analysis 128
7?4?1 Visual comparison method 128
7?4?2 Elution method 128
7?4?3 Thin-layer scanning method 129
7?5 Thin-layer chromatography progress 136
7?5?1 High-performance thin-layer chromatography 136
7?5? 2Reversed-phase thin-layer chromatography 136
7?5?3 Overpressure thin-layer chromatography 137
7?5?4 Rotating thin-layer chromatography 139
< p>7?5?5 Thin layer chromatography? Hydrogen flame ionization detection system (TLC? FID system) 1407?5?6 Ultra-thin layer chromatography 141
7 ?5?7 Thin layer chromatography? Photoacoustic spectroscopy technology 142
7?6 thin layer chromatography in biology Application in engineering analysis 144
7?6?1 TLC scanning method for the determination of FB?3 new preservative 144
7?6?2 TLC scanning method for the determination of compound Danshen The effective components of Salvia miltiorrhiza and Panax notoginseng 145
Chapter 8 Gas Chromatography 146
8?1 Overview 146
8?1?1 Gas Chromatography Separation Principle and process 146
8?1?2 Characteristics of gas chromatography 149
8?2 Basic theory 149
8?2?1 Basic concept 149
8?2?2 plate theory 150
8?2?3 rate theory 151
8?3 column and stationary phase 152
8?3?1 column 152
8?3?2 stationary phase 153
8?4 detector 156
8?4 ?1 The main performance indicators of the detector 156
8?4?2 Thermal conductivity detector 157
8?4?3 Hydrogen flame ionization detector 158
< p>8?4?4 electron capture detector 1598?4?5 nitrogen and phosphorus detector 160
8?5 qualitative and quantitative analysis method 160
< p>8?5?1 Qualitative analysis method 1618?5?2 Quantitative analysis method 161
The selection of 8?6 gas chromatography conditions 163
The choice of 8?6?1 column 163
The choice of 8?6?2 column temperature 164
The choice of 8?6?3 carrier gas and its flow rate 164
The selection of 8?6?4 carrier and fixative content 164
The choice of 8?6?5 other conditions 165
8?7 gas chromatography in bioengineering analysis 165
Determination of oxalic acid in 8?7?1 vegetables 165
8?7?2 Gas chromatography for direct determination of auxin 166
Chapter 9 High Performance Liquid Chromatography 168
9?1 Overview 168
9?1?1 Principles and Classification 168
9?1?2 Application range and limitations of high performance liquid chromatography 169
9?2 Basic theory of high performance liquid chromatography 171
9?2?1 chromatographic process 171
< p>9?2?2 Basic concepts 1729?2?3 Qualitative parameters 172
9?2?4 Column efficiency parameters 173
9? 2?5 phase equilibrium parameter 174
9?2?6 separation parameter 175
9?2?7 high performance liquid chromatography rate theory 176
9? 3High performance liquid chromatograph 177
9?3?1 Basic principles of high performance liquid chromatograph 178< /p>
The general steps of 9?3?2 high performance liquid chromatography analysis 183
The application of 9?4 high performance liquid chromatography in bioengineering analysis 185
9?4?1 amino acid analysis 185
9?4?2 peptide and protein separation analysis 186
9?4?3 nucleic acid and DNA analysis 188
< p>Chapter 10 Electrophoresis and High Performance Capillary Electrophoresis 19110?1 Overview 191
10?1?1 Electrophoresis 192
10?1 ?2High performance capillary electrophoresis 194
The basic theory of 10?2 electrophoresis and high performance capillary electrophoresis 196
The basic principle of 10?2?1 electrophoresis 196
< p>10?2?2 basic theory of capillary electrophoresis 19810?2?3 basic concept 200
10?2?4 theoretical efficiency and its expression method 203
p>10?3 Electrophoresis Apparatus and High Performance Capillary Electrophoresis Apparatus 205
10?3?1 Electrophoresis Apparatus 205
10?3?2 High Performance Capillary Electrophoresis Apparatus 208
p>10? 4 electrophoresis, high performance capillary electrophoresis analysis method analysis condition selection strategy 211
10? 4? 1 electrophoresis analysis method 211
10? 4? 2 high performance capillary Electrophoresis analysis method 214
10?5 application example 217
10?5?1 nucleic acid separation and analysis 218
10?5?2 sugar separation and detection 222
Chapter 11 Nuclear Magnetic Resonance Spectroscopy 224
11?1 Overview 224
11?1?1 Nuclear Magnetic Resonance Spectroscopy Development History and Research Status 224
The application of 11?1?2 NMR spectroscopy in the field of bioengineering 224
The basic theory and concept of 11?2 NMR spectroscopy 225
< p>11?2?1 Basic theory 22511?2?2 Several commonly used parameters in the NMR method 229
11?3 NMR spectrometer 230
11?3?1 RF transmitter system 230
11?3?2 probe 231
11?3?3 magnetic field system 231
11 ?3?4 Receiving detection system 232
11?3?5 Signal processing and control system 232
11?4 Basic experimental technology of nuclear magnetic resonance spectroscopy 233
< p>11?4?1 sample preparation 23311?4?2 standard reference sample 233
11?4?3 map analysis 234
11 ?4?4NMR commonly used pulse technology and its function 234
11?4?5 one-dimensional, two-dimensional Three-dimensional nuclear magnetic resonance experiment 238
11?5 application example 242
11?5?1NMR method to determine the three-dimensional structure of proteins and nucleic acids 242
11?5 ?2NMR determination of cell biofilm structure and intracellular pH246
11?5?3NMR determination of new biological drugs248
11?5?4NMR application in metabolomics 251
Chapter 12 Biological Mass Spectrometry 255
12?1 Overview 255
12?2 Principles of Mass Spectrometry 256
12?2? 1 Basic principles of mass spectrometry 256
12?2?2 Mass spectrometry terms and terms 257
12?3 Mass spectrometer 258
12?3?1 Mass spectrometry The basic structure of the instrument 258
The main performance indicators of the 12?3?2 mass spectrometer 262
The 12?3?3 mass spectrum and its interpretation 263
12 ?4 Biological mass spectrometry 264
12?4?1 Biological mass spectrometry 264
12?4?2 Laser desorption ionization and electrospray ionization mass spectrometry experiment 266< /p>
12?5 Biomass spectrometry application example 269
12?5?1 Peptide and protein analysis 269
12?5?2 Glycoprotein and oligosaccharide Analysis 272
12?5?3 Nucleotide analysis 274
Chapter 13 Bioassay 277
13?1 Overview 277
< p>13?2 Biological products biological test method 27813?2?1 bacteriological examination 278
13?2?2 pure bacteria test 279
< p>13?2?3 Microbial limit change test 28013?2?4 Mycoplasma test 280
13?2?5 Animal test 280
13 ?2?6 pyrogen test 282
13?2?7 endotoxin test 283
13?2?8 titer test 285
13?2 ?9 Bioassay statistical method 287
13?3 Biological assay method application example 289
13?3?1 Antibiotic microbiological assay method 289
13? 3?2 Insulin bioassay method 292
13?4 Molecular biology analysis method 292
13?4?1 Gene probe technology 292
13? 4?2 polymerase chain reaction 295
13?4?3 multisite enzyme electrophoresis 301
13?4?4 restriction enzyme analysis 302
13 ?4?5 Randomly amplified polymorphic DNA analysis 304
13?4?6 pulsed electric field gel electrophoresis Swimming analysis 306
13?4?7 Biochip analysis and detection technology 308
Chapter 14 Enzymatic analysis 315
14?1 Overview 315
14?2 Principles of enzymatic analysis 315
14?2?1 Enzyme characteristics and classification number 315
14?2?2 Principles of enzymatic analysis 317
14?3 Enzymatic analysis and application examples 323
14?3?1 Enzyme activity determination method 323
14?3?2 Enzyme activity determination Application examples of 326
Determination of 14?3?3 substrates and substrate analogs 328
Analysis and determination of 14?3?4 activators and inhibitors 332
p>Application of 14?3?5 immobilized enzyme in analytical flow system 333
14?3?6 Analysis of organic compounds 333
14?3?7 Analysis of inorganic compounds 334
The development trend of 14?4 enzymatic analysis in bioengineering 334
14?4?1 single cell analysis 334
Analysis of 14?4?2 enzyme as a marker 334
14?4?3 mimic enzyme analysis 334
Chapter 15 Immunoassay 336
15? 1Overview 336
15?2 Basic knowledge of immunoassay 336
15?2?1 Basic principles 336
15?2?2 antibody preparation 337< /p>
Identification of 15?2?3 antibody quality and evaluation of immunoassay method 340
15?3 Radioimmunoassay 342
15?3?1 label Preparation of antigen 342
The separation method of 15?3?2 free label and binding label 344
Drawing of 15?3?3 standard curve 345
15?3?4 basic steps of sample determination 346
15?4 enzyme-linked immunosorbent assay 346
15?4?1 principle 347
15 ?4?2 Several commonly used types of ELISA assay 348
15?4?3 enzyme conjugate 349
15?4?4 experimental conditions 352
15?5 Fluorescence immunoassay 353
15?5?1 Fluorescence polarization immunoassay 354
15?5?2 Fluorescence immunoassay 355
15?5?3 Quenching Fluorescence Immunoassay 356
15?6 New Technology and Trends in Immunoassay 356
15?6?1 Biotin? Avidin System 356
15?6?2 chemiluminescence enzyme immunoassay 356
15?6?3 enzyme cascade amplification 356
< p>15?6?4 Liposome Amplification Technology 35715?6?5 Immunoassay Reagent Strip 357
15?6?6 Immune Latex Detection Reagent 357
>15?6?7 genetic engineering antibody 357
15?6?8 automated and practical immunoassay 358
15?6?9 immunoassay and others Technology combination 359
15?6?10 Phase separation immunoassay based on temperature-sensitive hydrogel 359
15?7 Immunoassay application example 359
Application example of 15?7?1 RIA method in in vivo drug analysis 359
15?7?2 ELISA method application example 360
15?7?3FIA method application example 361
15?7?3FIA method application example 361
p>Chapter 16 Biosensor 363
16?1 Overview 363
16?1?1 Definition of biosensor 363
16?1 ?2 Development history of biosensors 363
16?1?3 Features of biosensors 364
16?1?4 Basic composition and working principle of biosensors 365
Classification of 16?1?5 biosensor 367
Application of 16?1?6 biosensor368
16?2 enzyme sensor 370
16?2?1 Structure of enzyme sensor 370
16?2?2 Classification of enzyme sensor 370
16?2?3 Several new enzyme sensors 371
16?2?4 enzyme sensor application 372
16?3 biological tissue sensor 373
16?3?1 animal tissue sensor 374
16?3?2 Plant tissue sensor 374
16?4 Cell and organelle sensor 374
16?4?1 Mitochondrial sensor 375
16?4? 2 Liver microsomal sensor 375
16? 4? 3 Chloroplast sensor 375
16? 5 Microbial sensor 375
16? 5? 1 Type of microbial sensor 375
16?5?2 Microbial Sensitive Film Preparation Technology 376
16?5?3 Electrochemical Microbial Sensor 376
16?5?4 Pressure Electrical high frequency impedance type microbial sensor 377
16?5?5 Fuel cell type microbial sensor 377
16?5?6 Other types of microbial sensor 378
16?5?7 Application of microbial sensor 378
16?6 immunosensor 380
16?6?1 Type of immunosensor 380
16? 6?2 Electrochemical immunosensor 380
16?6?3 Optical immunosensor 381
16?6?4 Piezoelectric crystal immunosensor 382
16?6? 5Surface Plasmon Resonance Type Immunosensor 383
16?6?6 Field Effect Transistor Biosensor 384
16?6?7 Light Addressable Potential Sensor 384
< p>16?6?8 receptor immunosensor 38416?6?9 immunochip 385
The problems and development prospects of 16?6?10 immunosensor 385
p>16?7 nucleic acid biosensor (gene sensor) 385
16?7?1 nucleic acid biosensor production and classification 385
16?7?2 DNA biosensor And DNA hybrid sensor 386
16?7?3 electrochemical DNA sensor 386
16?7?4 optical DNA biosensor 387
16?7? 5Surface plasmon resonance DNA sensor 388
16?7?6 Piezoelectric crystal DNA sensor 389
16?8 Introduction to several new biosensors 389
16?8?1 Optical fiber biosensor 389
16?8?2 Bionic biosensor 389
16?8?3 Molecularly imprinted biosensor 390
16? 9 Biosensor development status and application prospects390
Chapter 17 Progress in other new analysis and detection technologies 392
17?1 Application and development trend of modern analysis methods 392
17?1?1 Overview 392
17?1?2 Analysis method development trend 392
17?1?3 Analysis instrument development trend 393
17?2 Radioisotope Analysis Technology 395
17?2?1 Basic Principles 395
17?2?2 Measuring Instruments and Methods 396
17?2?3 Autoradiography 400
17?3 Chromatography Combined Technology 402
17?3?1 Chromatography Combined Technology Generation 402
17?3?2 Chromatography Hyphenated Interface Technology 403
17?3?3 Gas Chromatography Hyphenated Technology 403
17?3?4 Liquid Chromatography Technology 409
17?3?5 Introduction to other chromatographic coupling technologies 415
17?4 Flow injection separation analysis technology 415
17?4?1 Overview 415
17?4?2 Flow injection analysis device and basic operation principle 416
17?4?3FIA basic flow path 417
17 ?4?4FIA Online Concentration and Separation Technology 417
Application of 17?4?5 Flow Injection Technology 418
Chapter 18 New Technology of Sample Preparation 420
18?1 membrane separation technology 420
18?1?1 membrane separation technology principle 420
18?1?2 membrane separation technology classification 421
Characteristics of 18?1?3 membrane separation technology 421
Classification and properties of 18?1?4 membrane 421
18?1?5 membrane separation technology in bioengineering 422
18?2 Foam separation technology 422
18?2?1 Basic principles of foam separation 423
18?2?2 The effect of foam separation Factor 423
18?3 solid phase extraction technology 424
18?3?1 basic principles of solid phase extraction technology 424
18?3?2 solid Phase extraction method 425
Development direction of 18?3?3 solid phase extraction technology 425
Application of 18?3?4 solid phase extraction technology 425
18?4 solid phase microextraction technology 427
18?4?1 solid phase microextraction technology principle 427
18?4?2 solid phase microextraction device and extraction Method 427
Selection of 18?4?3 solid phase microextraction technology extraction conditions 428
18?4?4 application of solid phase microextraction technology 429
18?5 microwave assisted extraction technology (microwave dissolution method) 429
The principle of 18?5?1 microwave extraction 429
18?5?2 The basic operation method of microwave extraction 429
Selection of 18?5?3 extraction conditions 430
18?5?4 Features of microwave extraction 430
18?5?5 Microwave extraction technology Application 430
18?6 Headspace Gas Chromatography Technology 431
18?6?1 Static Headspace Chromatography Technology and Application 431
18?6?2 Purging? Trap Chromatography Technology 433
18?7 High Speed Countercurrent Chromatography Separation and Purification Technology 435
Principles of 18?7?1 High Speed Countercurrent Chromatography 435
Features of 18?7?2 High Speed Countercurrent Chromatography 435
18?7?3 High Speed Countercurrent Chromatography Instrument 436
18?7?4 Application of High Speed Countercurrent Chromatography 436
18?7?5 Application of high-speed countercurrent chromatography in the field of bioengineering 437
Reference 439