Methods of Enzymatic AnalysisPDF电子书下载

Section A:General 1

Ⅰ．Principles of Enzymatic Analysis 3

Ⅱ．Experimental Techniques 14

Ⅲ．Cell and Tissue Disintegration 43

Section B:Estimation of Substrates Preliminary Remarks 57

Ⅰ．Substances Involved in Carbohydrate Metabolism 59

1．Poly-,Oligo-and Disaccharides 59

a)Glycogen 59

b)Starch 63

c)Cellulose 64

d)Hemicelluloses 72

e)Heparin 79

Determination with ribonuclease 79

Determination with pyruvic kinase 84

f)Hyaluronic acid 87

Spectrophotometric method 87

Colorimetric method 90

g)Raffinose 93

h)Sucrose 99

i)Lactose 103

2．Monosaccharides and Derivatives 107

a)D-Sedoheptulose-7-phosphate 107

b)D-Sedoheptulose-1,7-diphosphate 111

Determination with aldolase 111

Determination with SDPase 113

c)D-Glucose 117

Determination with HK/G6P-DH 117

Determination with GOD/POD 123

d)D-Glucose-1-phosphate 131

e)D-Glucose-6-phosphate and D-fructose-6-phosphate 134

f)D-Gluconic acid 139

g)D-6-Phosphogluconate 143

h)D-Glucosamine 146

i)D-Glucosamine-6-phosphate 151

k)D-Fructose 156

l)D-Fructose-1,6-diphosphate 160

Determination with FDPase 160

m)L-Sorbose-6-phosphate 164

n)D-Sorbitol 167

o)myo-Inositol 171

p)D-Ribose-5-phosphate 175

q)L-Ribulose and L-arabinose 178

r)D-Ribulose 182

s)D-Ribulose-5-phosphate 186

t)D-Ribulose-1,5-diphosphate 188

u)L-Xylulose 191

v)D-Xylulose 194

Determination with DPN-xylitol dehydrogenase 194

w)D-Xylulose and D-xylose 196

Determination with D-xylose isomerase 196

x)D-Xylulose-5-phosphate 201

y)D-Erythrose-4-phosphate 205

z)L-Erythrulose 208

3．Three-Carbon Compounds 211

a)Glycerol 211

b)L-Glycerol-1-phosphate 215

c)D-Glycerate 220

d)D-3-Phosphoglycerate,D-2-phosphoglycerate,phosphoenolpyruvate 224

e)D-1,3-Diphosphoglycerate 234

f)D-2,3-Diphosphoglycerate 238

g)L-Glyceraldehyde-3-phosphate 241

h)Dihydroxyacetone 244

i)Dihydroxyacetone phosphate,D-fructose-1,6-diphosphate and D-glyceraldehyde-3-phosphate 246

k)Pyruvate 253

l)Hydroxypyruvate 260

m)β-Mercaptopyruvate 263

n)L-Lactate 266

Determination with LDH and DPN 266

Determination with yeast LDH 271

Determination with LDH and AP-DPN 275

o)D-Lactate 278

p)Methylglyoxal 283

4．Two-and One-Carbon Compounds 285

a)Ethanol 285

Determination with ADH and DPN 285

Determination with ADH and AP-DPN 287

b)Acetaldehyde 290

Determination with ADH 290

Determination with aldehyde dehydrogenase 292

c)Glycolaldehyde 297

d)Glyoxylic acid 300

e)Acetate 303

f)Formate 308

Ⅱ．Substances Involved in the Citric Acid Cycle 313

a)Citrate 313

Determination with citrase 313

b)Citrate and isocitrate 318

Determination with aconitase and isocitric dehydrogenase 318

c)α-Oxoglutarate 324

d)L-Malate 328

Determination with MDH and DPN 328

Determination with MDH and AP-DPN 332

e)Oxaloacetate 335

f)Succinate 340

g)Fumarate 344

Ⅲ．Substances Involved in Protein Metabolism 350

1．Proteins and Peptides 350

a)Characterization of peptides and proteins with enzymes 350

b)Glutathione 363

2．Amino Acids,Amines and Amides 367

a)D-Amino acids 367

b)L-Amino acids(Lys,Arg,Orn,Tyr,His,Glu,Asp) 373

c)L-Alanine 378

d)L-Aspartic acid and L-asparagine 381

e)L-Glutamic acid(and L-glutamine) 384

Determination with GIDH 384

Determination with GIDH and LDH 388

Determination with GIDH and AP-DPN 392

Determination with GPT 394

f)DL-Threonine 398

g)Urea 401

h)Creatine 407

Ⅳ．Substances Involved in the Metabolism of Fatty Acids,Lipids and Steroids 411

a)Polyunsaturated fatty acids 411

b)Lecithin 415

c)Acyl-CoA derivatives 419

Acetyl-CoA 419

Acetoacetyl-CoA 425

Acrylyl-CoA 429

Benzoyl-CoA 431

Butyryl-CoA and CoA derivatives of the higher saturated fatty acids 433

Crotonyl-CoA 437

L-(+)-β-Hydroxybutyryl-CoA 441

β-Hydroxy-β-methylglutaryl-CoA 445

β-Hydroxypropionyl-CoA 449

Malonylsemialdehyde-CoA 452

d)Acetoacetate 454

e)D-(－)-β-Hydroxybutyrate 459

f)Hydrolysis of steroid conjugates 462

g)20-Ketosteroids 477

Spectrophotometric method 477

Fluorimetric method 480

h)Steroid alcohols in urine 485

Ⅴ．Nucleosides,Purines and Pyrimidines,Coenzymes and Related Substances 491

1．Nucleosides,Purines and Pyrimidines 491

a)Adenosine 491

b)Hypoxanthine and xanthine 495

c)Uric acid 500

d)Inosine 502

e)Guanosine 505

f)Orotic acid 508

2．Coenzymes and Related Substances 512

a)Coenzyme A 512

Determination of CoA-SH and CoA-S-S-CoA with HOADH 513

Determination of CoA-SH with PTA 517

Determination of CoA-SH with TK 520

Determination of CoA-SH with CCE 523

b)Diphosphopyridine nucleotide 528

c)Reduced diphosphopyridine nucleotide 531

d)Triphosphopyridine nucleotide 535

e)Reduced triphosphopyridine nucleotide 537

f)Adenosine-5′-triphosphate 539

Determination with PGK 539

Determination with HK/G6P-DH 543

Determination by enzymatic fluorimetry 551

g)ATP and creatine phosphate 559

Determination with luciferase 559

h)Adenosine-5′-diphosphate and adenosine-5′-monophosphate 573

i)Adenosine phosphates 578

k)UDP-glucose,UDP-galactose,UTP,UDP-glucuronic acid 581

l)Flavine adenine dinucleotide 596

m)Flavine mononucleotide 599

n)Thiamine pyrophosphate 602

o)Pyridoxal phosphate 606

p)Creatine phosphate 610

Ⅵ．Other Substrates and Miscellaneous Substances 617

a)Organophosphorus insecticides 617

b)DDT 626

c)Inorganic peroxides 633

d)Nitrate 636

e)Magnesium in plasma 640

f)Pyrophosphate 644

Section C:Measurement of Enzyme Activity Preliminary Remarks 651

Ⅰ．The Importance of the Measurement of Enzyme Activity 651

a)In medicine 651

b)In food chemistry 713

c)In botanical and agricultural chemistry 720

Ⅱ．Methods 724

1．Aldolases 724

a)Fructose-1,6-diphosphate aldolase 724

Colorimetric determination with 2,4-dinitrophenylhydrazine 725

Determination with enzymatic auxiliary and indicator reactions 728

b)1-Phosphofructoaldolase 732

2．Dehydrogenases 736

a)Lactic dehydrogenase 736

b)Glucose-6-phosphate dehydrogenase 744

c)Glutamic dehydrogenase 752

d)Malic dehydrogenase 757

e)Sorbitol dehydrogenase 761

3．Esterases 765

a)Acetylcholinesterase 765

b)Cholinesterases 771

Colorimetric determination 772

Manometric determination 774

c)Lipase 776

d)Phosphatases(phosphomonoesterases) 779

Determination in serum with phenolphthalein diphosphate 779

Determination in serum with p-nitrophenylphosphate 783

Determination in milk with phenylphosphate 785

e)Glucose-6-phosphatase 788

f)Ribonuclease 793

Spectrophotometric method 794

Titrimetric method 797

4．Proteases 800

a)Chymotrypsin 800

Determination with casein as substrate 800

Determination with L-tyrosine ethyl ester as substrate 802

b)Trypsin 807

Determination with haemoglobin as substrate 808

Determination with casein as substrate 811

Determination with benzoylarginine ethyl ester as substrate 815

c)Pepsin 819

d)Peptidases 824

Acidimetric determination of dipeptidases 824

Colorimetric determination of carboxypeptidase 828

Colorimetric determination of leucine aminopeptidase in serum 830

5．Transaminases 837

a)Glutamate-oxaloacetate transaminase 837

Determination with MDH as indicator enzyme 837

Colorimetric determination with 2,4-dinitrophenylhydrazine 842

b)Glutamate-pyruvate transaminase 846

Determination with LDH as indicator enzyme 846

Colorimetric determination with 2,4-dinitrophenylhydrazine 851

6．Others 854

a)Amylase 854

b)Creatine phosphokinase 859

c)Galactose-1-phosphate uridyl transferase 863

d)β-Glucosidase 867

e)β-Glucuronidase 869

f)Glutathione reductase 875

TPNH-specific glutathione reductase 876

DPNH-specific glutathione reductase 877

g)Kallikrein 880

Determination of kallikreins 880

Determination of kallikrein inhibitors 883

h)Catalase 885

Spectrophotometric method 886

Determination by iodometric titration 888

Determination with the Katalaser 891

i)Peroxidase 895

k)Reductases 898

l)Inveftase 901

Determination in food chemistry 901

Determination in botanical and agricultural chemistry 904

m)Thromboplastin time 908

Determination according to Quick 909

＂Geigy＂micromethod 910

n)Urease 913

Titrimetric method 913

Colorimetric method 915

o)Xanthine oxidase 917

Ⅲ．Histochemical Detection of Enzymes 920

1．Introduction(Aims of the Method) 921

2．General 923

a)Principle techniques 923

b)Requirements 924

c)Sources of error 925

d)Range of application 925

e)Precautions necessary for the preservation of cell components 925

3．Special Methods 929

a)Methods involving precipitation of metallic salts 929

b)Methods employing azonaphthol dyes 943

c)Methods employing indigo dyes 949

d)Methods employing indoaniline and azamethine dyes 950

e)Methods employing tetrazolium salts 953

f)Detection of polysaccharide synthesis 609

Section D:Biochemical Reagents 967

Ⅰ．Enzymes 968

Ⅱ．Coenzymes and Substrates 1002

Ⅲ．Complete Reagent Kits 1034

Tables 1039

Conversion Table:Transmittance？Extinction 1041

Atomic Weights 1045

Index 1047