Contents. Section 1: Basic Laboratory Principles. 1. Laboratory Equipments, Glassware, Laboratory Hazards and General Laboratory Rules...

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1 Contents Section 1: Basic Laboratory Principles 1. Laboratory Equipments, Glassware, Laboratory Hazards and General Laboratory Rules...3 Laboratory Equipments 3 Laboratory Glassware 4 Laboratory Hazards 7 Laboratory Safety Procedure 7 Laboratory First Aid Laboratory Units, Basic Calculations and Preparation of Solutions...11 Units 11 Preparation of Molar and Normal Solutions 13 Preparation of Percent Solutions Acids, Bases, Alkalies, ph, Indicators and Buffers...15 Acids and Bases 15 Alkali 15 Indicators 15 Buffers 16 Section 2: Qualitative Experiments 4. Carbohydrates...21 Definition 21 Classification Reactions of Carbohydrates...23 Molisch s Test 23 Iodine Test 23 Benedict s Test 24 Barfoed s Test 25 Seliwanoff s Test 25 Osazone Test 25 Reactions of Monosaccharides 26 Reactions of Disaccharides 27 Scheme for Identification of Unknown Carbohydrates 28 Reactions of Polysaccharides 28

2 6. Proteins...30 Definition 30 Elemental Composition 30 Functions 30 Classification Color Reactions of Proteins...32 Protein and its Color Reactions Precipitation Reactions of Proteins...38 Methods of Precipitation Reactions 38 Significance of the Precipitation Reactions 39 Scheme for Identification of Unknown Proteins 41 Laboratory Manual for Practical Biochemistry 9. Reactions of Nonprotein Nitrogenous (NPN) Substances...42 Nonprotein Nitrogenous Substances Scheme for Identification of Physiologically Important Substances Urine Analysis (Normal Urine)...45 Analysis of Normal Constituents in Urine 45 Determination of Specific Gravity Analysis of Abnormal Constituents in Urine...50 Physical Characteristics 50 Chemical Constituents Inborn Errors of Metabolism...54 Screening Tests for Inborn Errors of Amino Acid Metabolism 54 Screening Tests for Inborn Errors of Carbohydrate Metabolism Analysis of Gastric Juice...58 Tests for Normal Constituents 58 Tests for Abnormal Constituents Analysis of Saliva...61 Collection of Saliva Preparation of Hemin Crystals...63 Preparation of Hemin Crystals Sample Collection and Preservation...64 Blood Sample Collection and Preservation 64 Urine Collection and Preservation 65 xii

3 Section 3: Quantitative Experiments 18. Estimation of Blood Sugar by Folin-Wu Method...69 Principle 69 Procedure 69 Calculation 70 Clinical Aignificance 71 Other Methods Estimation of Blood Glucose by O-Toluidine Method...72 Principle 72 Procedure 72 Calculation Estimation of Blood Urea by Diacetyl Monoxime (DAM) Method...74 Principle 74 Procedure 74 Calculation 75 Clinical Significance 75 Other Methods Estimation of Serum Total Protiens, Albumin and Albumin/Globulin Ratio by Biuret Method...77 Principle 77 Procedure 77 Calculation 78 Clinical Significance 78 Contents 22. Estimation of Serum Albumin by Dye-Binding Method...80 Principle 80 Procedure 80 Calculation 80 Other Methods Estimation of Serum Inorganic Phosphate by Fiske and Subba Row Method...82 Functions of Phosphorus 82 Principle 82 Procedure 83 Calculation 83 Clinical Significance 83 Other Method 84 xiii

4 24. Estimation of Serum Cholesterol by Zak s Method...85 Principle 85 Procedure 85 Calculation 86 Clinical Mignificance 86 Other Methods Estimation of Serum Uric Acid by Caraway Method...87 Principle 87 Procedure 87 Calculation 88 Clinical Significance 88 Other Methods 88 Laboratory Manual for Practical Biochemistry 26. Estimation of Serum Bilirubin by Malloy and Evelyn Method...89 Principle 89 Procedure 89 Calculation 90 Clinical Significance Estimation of Serum Creatinine by Jaffe s Alkaline Picrate Method...92 Creatine 92 Principle 92 Calculation 93 Clinical Significance Estimation of Vitamin C (Ascorbic Acid) in Plasma by Sauberlich Method...94 Principle 94 Procedure 94 Calculation 94 Clinical Sgnificance Estimation of Vitamin A in Serum by Bessey Method...96 Principle 96 Procedure 96 Calculation 96 Clinical Significance 96 xiv 30. Estimation of Serum Transaminases by Reitman and Frankel Method...97 Principle 97 Procedure 97 Calculation 98 Clinical Significance 98 Other Methods 99

5 31. Estimation of Serum Alkaline Phosphatase by Kind and King Method Principle 100 Clinical Significance Estimation of Serum Calcium by O-Cresolphthalein Complexone Method Principle 102 Calculation 103 Clinical Significance 103 Other Methods Estimation of Urine Creatinine By Jaffe Alkaline Picrate Method Principle 104 Procedure 104 Calculation 104 Clinical Significance Estimation of Urinary Proteins by Turbidimetric Method Principle 106 Procedure 106 Calculation 106 Other Methods 107 Contents Section 4: Clinical Biochemistry Techniques and Recent Aspects of Clinical Biochemistry 35. Colorimetry General Principle and Procedure Principle 111 Procedure 113 Calculation 113 Applications of Colorimetric Assays Potentiometry Indicator Electrodes 115 ph Meter 115 Ion-Selective Electrodes Flame Photometer General Principle and Procedure Flame Photometry 118 xv

6 38. Electrophoresis General Principle and Procedure Electrophoresis Chromatography General Principle and Procedure Chromatography Automation in Clinical Biochemistry Types of Analyzers Quality Control in Clinical Biochemistry Investigation of Clinical Laboratory Oral Glucose Tolerance Test Assessing the Status of Glucose Tolerance 134 Laboratory Manual for Practical Biochemistry 43. Case Reports Normal Values of Biochemical Parameters Section 5: Reagent Preparations 45. Reagent Preparations Reagents for Qualitative Experiments 153 Reagents for Quantitative Experiments 155 Index xvi