Simplified Practical Manual of Biochemistry Javin Bishnu Gogoi
INDEX
Page numbers followed by f refer to figure, and t refer to table
A
Acetic acid 23, 180
Acetone 65
Acid 4, 5
splashes 4
Acrolein test 73
Adenosine deaminase deficiency 154
Adler's test 92, 93
Agar 24, 109
Agarose gel 109
Air displacement micropipettes 9
Alanine aminotransferase 171
Albumin 66, 122, 157
estimation of 162
Alcohol 65
Alkali 4, 5
splashes 4
Alkaline 29
copper reduction methods 129
phosphatase 176
Alkaloids 29
Aluminium chloride 24
Amino acids 110
sulfur-containing 61, 62
Ammonia 24, 122
Ammonium
hydroxide 24
molybdate solution 24
oxalate 25
Amniocentesis 31
Amniotic
fluid 31, 32
sac 31
Amylase solution 25
Amylopectin 54
Amylose 54
Analbuminemia 164
Anemia 181
Apparatus 109
Arginine 61, 62
Aromatic amino acids 61, 62
Arterial blood puncture 31
Asatoor and King method 128
Ascitic fluid 31, 32
Aspartate aminotransferase 171
Automated biochemistry analyzers, types of 99
Automatic chemistry analyzers 15
Automation, advantage of 99
B
Bacterial meningitis 123
Barfoed's reagent 25
Barfoed's test 41, 4345, 47, 49, 51, 52
Barium
chloride 25
hydroxide 24
Beaker 8, 9f
Beer-Lambert law 105, 106f
Bence Jone's protein 109
Benedict's qualitative test 91, 93
Benedict's solution 25
Benedict's test 40, 41f, 4345, 47, 49, 5153
Benedict's uric acid test 81, 86
Benzidine test 92, 93
Berthelot reaction 139
Bial's reagent 25
Bial's test 46, 47
Bile
pigments 90, 93
salts 90, 91, 93
Bilirubin, standard 168
Biochemical tests 3
Biuret
method 157
reagent 25, 157
test 59, 61, 62
Blood 90, 93
cells 123
glucose, estimation of 127
sample, collection of 32, 127
Body fluids, types of 32
Bone diseases 158
Brady's reagent 26
Brain tumor 123
Bulb pipettes 9
Bunsen burner 7, 7f, 18
Burns, severe 141
C
Calcium 84, 122
chloride solution 26
hydroxide 24
solution 26
Capillary electrophoresis 109, 111
Carbohydrates 37, 42, 42t, 93
detection of 37
Carcinoma bladder 141
Carway's method 153
Cellulose 54
acetate membrane 109
Centrifuge 12, 13f
types of 13
Cerebrospinal fluid 31, 32, 121, 122
analysis of 121
chloride 123
gamma globulin 123
glucose 122
normal 122
pressure 122
protein 122
sample, collection of 122
spaces 121f
Chloride 84, 122, 123t
Cholesterol 74
oxidase-peroxidase method 181
standard 26, 180
tests of 74
working standard 26
Chromatography 114
types of 115
Collagen diseases 158
Color Maumene's test 71
Colorimeter 15, 15f
Colorimetric method 139
Continuous tubular centrifuges 13
Copper sulfate solution 26
Creatine phosphate 148
Creatinine clearance test 148
Crystalline structure 42
Cubital vein 31
Cumulative sum control chart 100, 101
Cystic kidneys, congenital 141
D
Dementia 123
Dextrins 54
Diabetes
alimentary 133
diagnosis of 133
gestational 133
mellitus 123, 181, 187
Diacetyl monoxime 139
method 139
Diarrhea, severe 140
Diazo reaction 61, 62, 167
Digestive glycosuria 133
Digital weighing machine 12f
Dihydroxy acetone phosphate 185
Dilute iodine solution 27
Diphenylamine 26
Direct photometric methods 159
Dunstan's test 73
Dye-binding method 159, 162
E
Ehrlich's test 92, 93
Electrophoresis 108, 162
principle of 108, 108f
types of 109
uses of 109
Emulsification test 72
Encephalitis 123
Enzymatic method 127, 153, 181
Epilepsy 123
Ethereal sulfate 85
Ethylenediaminetetra acetic acid 33
Eye burns 4
F
Fasting
blood sugar 127
glycemia, impaired 133
plasma glucose 133
serum sample 186
Febrile seizure 123
Fehling's solution 26
Fehling's test 40
Ferric chloride 180
reagent 181
Ferricyanide method 128
Fevers 141
Flat glucose tolerance curve 136
Fluids, synovial 31
Fluorescence 110
Foam test 92, 93
Folin-Wu method 128
modified 128
Formalin solution 27
Fouchet's test 91, 93
Free fatty acids 72
Friedewald's equation 182
Froehde's reagent 29
Fructosazone 42
Funnel 10f
G
Galactosazone 42
Galactose 48
Galvanometer 106
Gamma globulin 122
electrophoresis 111
Gas centrifuges 13
Gastric aspiration 31
Gaussian chart 100
Gaussian curves, normalized 101f
Gel
electrophoresis 111
filtration chromatography 116, 116f
Gelatin 29
Generalized tonic-clonic seizure 123
Gerhardt's test 91, 93
Glass
micropipettes 9
pipette 10f
probes, storage of 103
Globulin 66, 157
free filtrate 163
Glomerular filtration rate 139
Glomerulonephritis, acute 141
Glucosazone 42
Glucose 91, 122
oxidase method 127, 128
solution 27
standard 128
tolerance
curve 134, 135
test 132
Glucosides 29
Glycerokinase 186
Glycerol 73, 185
Glycerophosphate-oxidase method 185
Glycogen 38, 54
Glycoprotein 110
Glycosuria, alimentary 133, 135
Gmelin's test 91, 93
Gout 154
Grease spot test 72
Guaiac solution 27
Guaiac test 92, 93
H
Hay's sulfur test 91, 93
Heat coagulation test 64, 90, 93
Heavy metal ions 65
Heller's nitric acid test 90, 93
Hemolysis 33
Hemorrhage 141
subarachnoid 123
Heparin 33
Hexokinase method 127, 128
High performance liquid chromatography 117, 117f
High voltage electrophoresis 109, 111
Histidine 61, 62
Hopkins-Cole reaction 27, 6062
Hot air ovens 14, 14f
Hubl's iodine 27
Hydrocephalus 123
Hydrochloric acid 20, 23
Hydrogen peroxide 27
Hydrolysis 53
test 42, 54
Hyperalbuminemia 163
Hyperchloremic acidosis 123
Hypercholesterolemia 181
Hyperglycemia, causes of 129
Hyperlipidemia 186
Hyperthyroidism 181
Hypertriglyceridemia 186
Hyperuricemia 154
Hypoalbuminemia 163
Hypocholesterolemia 181
Hypoglycemia, causes of 129
Hypoproteinemia 158
Hypothyroidism 181, 187
Hypotriglycerolemia 187
Hypouricemia 154
I
Immunoelectrophoresis 109, 111
Incubation 181
Infection
acute 158, 181
chronic 109, 158
Infectious specimen, accidental swallowing of 4
Inflammatory polyneuropathy, chronic 123
Inhalation anthrax 123
Inorganic
phosphates 122
sulfate 85
Intestinal obstruction, acute 140
Inulin 54
Inversion test 42, 53
Iodine 27
solution 27
test 38, 39f, 4345, 47, 49, 5154, 92, 93
Iodoacetate 33
Iron, determination of 30
J
Jaffe's alkaline picrate method 148
Jaffe's reaction, disadvantage of 149
Jaundice, obstructive 181
Joints, synovial 31
K
Kala-azar 158
Ketoacidosis, diabetic 134, 140
Ketone bodies 90, 91, 93
Kjeldahl's method 159
L
Lactate 122
Lactosazone 42, 50
Lambert's law 105
Lead sulfide test 61, 62
Lesch-Nyhan syndrome 154
Leukemia 154
Leukocyte count 122
Levey-Jennings chart 100, 101f
Liebermann-Burchard test 74
Lime water 26, 27
Lipemic sample 186
Lipids 73
qualitative analysis of 71
Lipoprotein 110
lipase 186
Liver
cirrhosis of 109
disease
advanced 154
chronic 158
Lowry method 159
Lumbar puncture 31
M
Macroglobulinemia 158
Malabsorption syndrome 181
Malignant hypertension 141
Malloy and Evelyn method 167
Maltosazone 42, 50
Mass concentration 20
Meningitis, tuberculous 123
Meniscus, reading of 16
Mercuric chloride 27
Methylene blue stain 28
Microfluidic pipette 9
Micropipette 10f
Millon's test 6062
Molal solution 21
Mole fraction 21
Molisch's reagent 28
Molisch's test 37, 38f, 4345, 47, 49, 5154
Mucic acid test 48, 49
Multiple myeloma 109, 158
N
Nephelometric methods 159
Nephritic syndrome 109
Nessler's method 139
Nessler's reagent 28
Neutral ferric chloride solution 28
Nile pigments 91
Ninhydrin
solution 28
test 59, 61, 62
Nitric acid 23
Nitroprusside
reaction 6062
solution 28
Non-enzymatic method 128, 153
Nonprotein nitrogen 139
Normal pressure hydrocephalus 123
Nucleic acids 110
Nucleoproteins 110
O
Oral glucose challenge test 133
Organic sulfate 85
Ortho-toluidine method 129
Osazone 42
mixture 28
test 4245, 47, 49, 51, 52
Ostwald-Folin pipette 9
O-toluidine method 128
Oxalates 32, 33
P
Pancreatitis 187
Paper
chromatography 115, 116f
electrophoresis 109, 109f, 110t
Pasteur pipettes 9
Pauly's test 61, 62
P-chlorophenol 186
Pentose 46
Pericardial fluid 31, 32
Peritoneal fluid 31
Peroxidase 186
pH meter 15, 15f, 103, 104f
operation of 103
principle of 103
Phenylhydrazine test 42
Phloroglucinol-HCl test 48, 49
Phosphoric acid 23
Phosphotungstic acid 154
test 81, 86
Photo colorimeter, parts of 106, 106f
Photometry 105
Picric acid test 42
Pipes buffer 186
Pipette 9
Pituitary tumor 123
Plasma 31, 32
Poisonous reagents, accidental swallowing of 4
Polyacrylamide gel electrophoresis 109
Polycythemia 154
Polyethylene glycol 4
Polysaccharide 54, 110
Postprandial blood sugar 127
Potassium
chloride 28
hydroxide 24
Pressure 122
Prostate, enlargement of 141
Protein 64, 90, 110, 122, 123t
color reactions of 59
estimation 159
solution, standard 157
Pulsed field gel electrophoresis 109, 111
Pyelonephritis, chronic 141
Pyloric stenosis 140
R
Random blood sugar 127
Reduction test 54, 73
Refractive index 71
Renal
failure 123, 141
glycosuria 133, 134
tubular defect 154
Reye syndrome 123
Rothera's test 91, 93
Rotor design 13
Ryle's tube 31
S
Sakaguchi test 6062
Saliva 31, 32
Salkowski test 74
Salt precipitation method 162
Saponification test 72, 73
Schiff's reagent 28
Schiff's test 81, 86
Screen centrifuges 13
Seliwanoff's reagent 28
Seliwanoff's test 41, 4345, 47, 49, 51, 52
Serum 31, 32
albumin, estimation of 162, 163
alkaline phosphatase, estimation of 176
bilirubin, estimation of 167
creatinine, estimation of 148
glutamate
oxaloacetate transaminase 171
pyruvate transaminase 171
high density lipoprotein cholesterol, estimation of 182
low density lipoprotein cholesterol, estimation of 182
total proteins, estimation of 157
triglycerides, estimation of 185
Shock 141
Silver nitrate 28
Skin 4
puncture 31
Sodium
carbonate 153, 154
chloride 28
citrate 33
fluoride 32, 33, 127
hydrogen carbonate 24, 29
hydroxide 24
hypobromite
solution 29
test 77, 78, 86
pyrogallol 29
Solubility test 71, 72
Spectrophotometer 106
Spirit lamp 8, 8f
Starch 54
solution 29
Stock solution 27
Stomach 31
Strip method 128
Subarachnoid space 31
Sucrose 29
Sugar 90, 91
Sulfanilic acid 29
Sulfhydril groups 6062
Sulfomolybdic acid 29
Sulfosalicylic acid 29
test 90, 93
Sulfur reaction 60, 61
Sulfuric acid 24, 29, 180, 181
Surface tension test 91, 93
Sweat 31, 32
T
Tannic acid 29
Tauber's benzidine test 46
Tauber's reagent 29
Test tube 6, 6f
holder 6f, 7
position of 18
stand or rack 7, 7f
Thermal test 71
Thin layer chromatography 115, 116f
Thyrotoxicosis 141
Tollen's reagent 29
Tollen's test 48, 49
Total serum cholesterol, estimation of 180
Triacylglycerol 185
Triglycerides 185
Tungstic acid reagent 153
Turbidimetric methods 159
Two-dimensional gel electrophoresis 111
Tyrosine 61, 62
U
Ulcerative colitis 140
Ultracentrifuges 13
Urea
estimation of 139
qualitative reactions of 77
Urease test, specific 77, 78, 86
Urethral strictures 141
Uric acid 153
estimation, method of 153
qualitative reactions of 80
standard 154
Uricase enzymatic method 153, 154
Urinary
ammonia 86
creatinine 86, 148
tract 141
urea 85
estimation 144
uric acid 86
Urine 31
abnormal 90
analysis, normal 83
inorganic constituents of 84
normal 83
organic constituents of 85
preservatives 33, 83
sample 32
Urinometer 12, 12f
Uristix method 127
Urobilinogen 92, 93
V
Viral infection 123
Viscosity 71
Vomiting 140
von Gierke's disease 154, 187
X
Xanthine oxidase deficiency 154
Xanthoproteic test 6062
Z
Zak's method 180
Zak's test 74
Zimmermann-Reinhardt reagent 30
Zinc chloride solution 30
×
Chapter Notes

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1INTRODUCTION
  • Clinical Biochemistry Laboratory: An Introduction
  • Introduction of Basic Instruments used in a Biochemistry Laboratory
  • Types of Reagents or Solutions used in a Biochemistry Laboratory
  • Preparation of Some Common Reagents used in Biochemistry Laboratory
  • Sample Collection, Use of Preservatives and Anticoagulants
2

Clinical Biochemistry Laboratory: An IntroductionCHAPTER 1

The Clinical Laboratory is a place where blood, different body fluids (urine, saliva, cerebrospinal fluid, pus, gastric juice, peritoneal fluid, pleural fluid, etc) and other biological specimens are analyzed to know the well-being of a person. The tests are performed either manually or by using special instruments. The precisely measured results are calculated and interpreted to know the disease condition of a patient.
The Clinical Biochemistry is that branch of Laboratory Medicine which uses chemical analysis of various body fluids to study the levels of body constituents in health and disease conditions. Though blood is the most common sample analyzed for biochemical tests, urine and other body fluids are also analyzed frequently. The test results are analyzed and interpreted by the physician for diagnosis, treatment and monitoring of disease conditions. The results are also used to counsel the patient in preventive medicine.
 
BASIC USES OF BIOCHEMICAL TESTS
Biochemical investigations are involved, to varying degrees, in every branch of clinical medicine.
  • The results of biochemical tests may be of use in diagnosis and in the monitoring of various disease conditions.
  • Biochemical tests may also be of value in screening of disease or in assessing the prognosis once a diagnosis has been established.
  • The biochemistry laboratory is often involved in research into the biochemical basis of diseases and in clinical trials of new drugs.
 
SAFETY MEASURES IN LABORATORY
The minimum precautions to be taken in biochemistry laboratory include:
  • All chemicals and reagents should be well labeled and hazard noted.
  • Reagent bottles should not be held by the neck and should be kept on shelves, not higher than eye level.
  • Mouth pipetting must never be done for any reagent, solution, acid or alkali while working in the laboratory.
  • Small measuring cylinders should be used for measuring acids and alkalis. If more accurate volume is required, then a pipette plugged with non absorbent cotton wool or with rubber tube attached should be used.
  • Exposure should be minimum for those organic solvents which have toxic properties.
  • Never add water to acid but always add acid to water.4
  • Every sample should be considered as a contaminated sample; so proper handling should be done with utmost care and precaution.
  • Wash your hand with plenty of water after practical.
  • Always wear white apron while working in the laboratory.
  • Always wear shoes while working in the laboratory.
  • Never eat anything in the laboratory while working.
  • Hair should be tied neatly.
 
BASIC FIRST AID IN LABORATORY
Every biochemistry practical laboratory should have a first aid kit containing cotton wool, gauze pieces, spirit, bandage, medicinal adhesive tape, scissors, tincture iodine (1 g iodine in 95% alcohol), disinfectant solution, sterile normal saline, sodium carbonate, 5% (v/v) acetic acid, 8 % (w/v) magnesium hydroxide (milk of magnesium), 5% (w/v) soap solution.
The following first aid measures should be implemented if and when necessary:
 
Contacts with Corrosive Chemicals and Reagents
 
Acid Splashes on Skin
  • Wash the affected skin with plenty of tap water.
  • Bathe the affected skin with cotton wool soaked in 5% solution of sodium carbonate.
  • Rinse the affected skin in mild detergent.
  • Seek medical help.
 
Alkali Splashes on Skin
  • Wash the affected skin with plenty of tap water.
  • Bathe the affected skin with cotton wool soaked in 5% acetic acid.
  • Seek medical help.
 
Contact with Phenol
  • Wash the affected skin with plenty of tap water.
  • Use polyethylene glycol (PEG) mixed with water for further irrigation of affected skin.
  • Seek medical help.
Eye burns by alkali or acids need immediate attention. If it happens:
  • Immediately wash the eyes with plenty of tap water.
  • Rinse the eyes with sterile saline.
  • Consult an ophthalmologist immediately.
 
Accidental Swallowing of Poisonous Reagents
  • Spit it out immediately.
  • Rinse the mouth promptly with tap water.
  • Induce vomiting by drinking warm salt water (one table spoon of common salt in glassful of warm water).
  • Send to emergency immediately.
 
Accidental Swallowing of Infectious Specimen
  • Spit it out immediately.
  • Wash the mouth with dilute antiseptic lotion.
  • Rinse the mouth thoroughly with tap water.
  • Seek medical help.5
 
Accidental Swallowing of Corrosive Reagents
 
Acids
  • Promptly rinse the mouth with tap water.
  • Antidote such as 5% soap solution, 8% magnesium hydroxide or egg mixed with about 500 mL of water, can be used orally to neutralize the acids.
  • Seek medical help immediately.
 
Alkalis
  • After rinsing the mouth with tap water, antidote such as lemon juice or 5% acetic acid can be taken orally to neutralize the alkalis.
  • Seek medical help immediately.
 
Injuries caused by broken glass
  • Wash the wound immediately with a disinfectant solution.
  • Send to emergency after primary dressing in the laboratory.
 
Burns in the Laboratory
  • Immediately bathe the affected area in cold water.
  • Cover the burnt area with sterile dressing as early as possible.
  • If the victim is on fire, put out the fire by using any material such as sheets, towels or coats.
  • Remove smoldering clothes as quickly as possible.
  • Seek emergency medical help immediately.
REVIEW QUESTIONS
  1. What is the clinical importance of analyzing various biochemical parameters in medical sciences?
  2. What are the different safety measures that are to be followed while working in a laboratory?
  3. Enumerate the items that a first aid kit present in a student or clinical laboratory should contain.
  4. What are the first aid measures to be followed if a person faces acid splashes on skin?
  5. What are the first aid measures to be followed if a person faces alkali splashes on skin?
  6. What are the first aid measures to be followed if a person faces acid or alkali burn in eyes?
  7. What are the first aid measures to be followed if a person faces contact with phenol?
  8. What are the first aid measures to be followed if a person accidentally swallows a poisonous reagent?
  9. What are the first aid measures to be followed if a person accidentally swallows an infectious specimen?
  10. What are the first aid measures to be followed if a person accidentally swallows an acid?
  11. What are the first aid measures to be followed if a person accidentally swallows an alkali?
  12. What are the first aid measures to be followed if a person faces injury by a broken glass?
  13. What are the first aid measures to be followed if a person faces a burn in laboratory?