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Practical Biochemistry for BDS
Purvi Purohit
SECTION 1: INTRODUCTION TO PRACTICAL BIOCHEMISTRY
CHAPTER 1:
Biochemistry in Dentistry
CHAPTER 2:
A Peep into the Biochemistry Laboratory
COMMON LAB EQUIPMENT AND INSTRUMENTS
Common Lab Glassware
Samples
Anticoagulants
CHAPTER 3:
Quality Control and Precision of Results
IMPORTANCE OF QUALITY CONTROL IN CLINICAL PRACTICE
CHAPTER 4:
How to Work in a Biochemistry Laboratory
INSTRUCTIONS FOR WORKING IN A BIOCHEMISTRY LABORATORY
SECTION 2: QUALITATIVE EXERCISES
CHAPTER 5:
Carbohydrate Analysis
DEFINITION
Classification
TESTS FOR ANALYSIS OF CARBOHYDRATES
1. Molisch’s Test
Reagents
Principle
Procedure
Precautions
2. Reduction Tests
a. Fehling’s Test
Reagents
Procedure
Drawbacks
b. Benedict’s Test
Reagents
Principle
Procedure
Advantages of Benedict’s Test
Clinical Importance of the Benedict’s Test
Standard Way to Perform the Benedict’s Test with Urine Sample
3. Test for Detecting Monosaccharides
Barfoed’s Test
Reagents
Principle
Procedure
Precautions
4. Test for Keto—Sugars
Seliwanoff’s Test
Reagents
Principle
Procedure
5. Tests for Disaccharides
Inversion Test for Sucrose
Reagents
Procedure
6. Osazone Test
Reagents
Osazone Mixture
Glacial Acetic Acid
Principle
Procedure
TESTS FOR POLYSACCHARIDES
7. Iodine Test
Reagents—I2 Solution
Principle
Explanation
Precaution
Procedure
8. Half and Full Saturation Test
Reagents
Solid Ammonium Sulfate
Principle
Procedure
Half Saturation
Full Saturation
IDENTIFICATION OF UNKNOWN CARBOHYDRATE
VIVA VOCE
CHAPTER 6:
Lipid Analysis
DEFINITION
Classification
ANALYSIS OF LIPIDS BASED ON PHYSICAL PROPERTIES
I. Solubility Test
Principle
Procedure
II. Grease Spot Test
Reagents
Ether
Principle
Procedure
III. Emulsification Test (Fig. 6.1)
Reagents
Principle
Procedure
ANALYSIS OF DERIVED LIPIDS
Principle
Principle
Procedure
Principle
Procedure
Principle
Procedure
VIVA VOCE
CHAPTER 7:
Protein Analysis
DEFINITION
Classification
GROUP SPECIFIC TEST OF PROTEINS
i. Biuret Test
Reagents
Principle
Procedure
ii. Ninhydrin Test (for α-Amino Acids)
Reagents
Principle
Explanation
Practical Importance
Procedure
COLOR REACTIONS FOR IDENTIFICATION OF VARIOUS AMINO ACIDS
iii. Xanthoproteic Test (for Aromatic Amino Acids)
Reagents
Principle
Procedure
iv. Millon Nasse’s Test (for Tyrosine)
Reagents
Principle
Procedure
v. Lead Sulfide Test (for Sulfur Releasing Amino Acids – Cystine and Cysteine)
Reagents
Principle
Explanation
Procedure
vi. Aldehyde Test (for Tryptophan)
Reagents
Principle
Procedure
PRECIPITATION OF PROTEINS
vii. Heat Coagulation Test
Principle
Procedure
viii. Heller’s Nitric Acid Test
Reagents
Principle
Procedure
ix. Half and Full Saturation Test
Reagents
Principle
Procedure
Half Saturation Test
Full Saturation Test
x. Precipitation by Heavy Metals Test
Reagents
Principle
Practical Importance
Procedure
xi. Precipitation by Alkaloidal Reagent (Sulfo salicylic Acid)
Reagents
Principle
Clinical Importance
Procedure
xii. Precipitation at Isoelectric pH
Reagents
Principle
Procedure
xiii. Neumann’s Test
Reagents
Principle
Procedure
Explanation
IDENTIFICATION OF UNKNOWN PROTEIN
VIVA VOCE
SECTION 3: CLINICAL BIOCHEMISTRY
CHAPTER 8:
Gastric Juice Analysis
ANALYSIS OF GASTRIC JUICE
Method of Collection of Gastric Juice
DETERMINATION OF FREE AND TOTAL ACIDITY
Reagents
Principle
Procedure
OBSERVATION TABLE
Calculation
Normal Values
Interpretation
General Causes
VIVA VOCE
CHAPTER 9:
Colorimetry
PHOTOMETRY
Theory of Colorimetry
Photoelectric Colorimeter
Significance of Blank Solution
Choice of Filters
Spectrum
Instruction of Operating of Instrument
VIVA VOCE
CHAPTER 10:
Blood Sugar
ESTIMATION OF BLOOD GLUCOSE BY ORTHOTOLUIDINE METHOD OF DUBOWSKI, (1962)
Principle
Reaction
Reagents
Procedure
Calculation
ESTIMATION OF BLOOD GLUCOSE BY GLUCOSE OXIDASE OR PEROXIDASE
Reagent
Principle
Procedure
Calculation
Normal Value and Clinical Significance
VIVA VOCE
CHAPTER 11:
Total Protein and A:G Ratio
ESTIMATION OF TOTAL PROTEIN BY BIURET METHOD
Reagents
Principle
Procedure
Calculation
Normal Values and its Significance
ESTIMATION OF ALBUMIN BY BCG METHOD
Reagents
Principle
Reaction
Procedure
Calculation
Normal Values and Significance
Serum Globulin
Normal Value and its Significance
VIVA VOCE
CHAPTER 12:
Serum Urea
ESTIMATION OF SERUM UREA BY BERTHELOT METHOD
Principle
Reagents
Procedure
Calculation
Normal Value and its Significance
VIVA VOCE
CHAPTER 13:
Serum Creatinine
ESTIMATION OF SERUM CREATININE (BY JAFFE’S METHOD)
Reagents
Principle
Procedure
Calculation
Normal Values and its Significance
Estimation of Urinary Creatinine
VIVA VOCE
CHAPTER 14:
Serum Lipid Profile
ESTIMATION OF SERUM CHOLESTEROL (ENZYMATIC METHOD)
Method: Cholesterol Oxidase Peroxidase Enzymatic (Allain CC et al 1974).
Reagents
Principle
Procedure
Calculation
Normal Values
ESTIMATION OF HDLC
Method: Mg–Phosphotungstic Acid Precipitation (Gordon T et al 1977)
COMPONENTS OF PRECIPITATING REAGENTS
Principle
Precipitation Procedure
Calculation
Normal Values
ESTIMATION OF TRIGLYCERIDES
Method
Reagents
Principle
Procedure
Calculation
Normal Values
VERY LOW DENSITY LIPOPROTEIN CHOLESTEROL
Normal Values
LOW DENSITY LIPOPROTEIN CHOLESTEROL
Normal Values
Limitations of Friedewald’s Equations
Clinical Interpretations
VIVA VOCE
CHAPTER 15:
Estimation of Sugar in Urine (Benedict’s Quantitative Method)
PRINCIPLE
Reagents
Procedure
Observation
Calculation
VIVA VOCE
SECTION 4: URINE ANALYSIS
CHAPTER 16:
Analysis of Normal Urine
TEST FOR ORGANIC CONSTITUENTS
Tests for Urea
Urease Test
Hypobromide Test
Tests for Creatinine
Jaffe’s Test
Tests for Uric Acid
Murexide Test
TEST FOR INORGANIC CONSTITUENTS
VIVA VOCE
CHAPTER 17:
Analysis of Abnormal/Pathological Urine
PROTEINS
DETECTION OF PROTEINS IN URINE
DETECTION OF SUGAR IN URINE
True Glycosuria
Renal Glycosuria
Alimentary Glycosuria
DETECTION OF KETONE BODIES
Rothera’s Test
DETECTION OF BILE PIGMENTS AND BILE SALTS
Detection of Bile Pigments in Urine by Gmelin’s Test
Principle
Procedure
Detection of Bile Salts in Urine by Hay’s Sulfur Test
Principle
Procedure
DETECTION OF BLOOD
Benzidine Test
Reagents
Principle
Procedure
VIVA VOCE
SECTION 5: APPLICATIONS OF CLINICAL BIOCHEMISTRY
CHAPTER 18:
Liver Function Tests
TESTS BASED ON BILIRUBIN METABOLISM
Principle
Clinical Interpretations
Fecal and Urinary Urobilinogen
TESTS BASED UPON METABOLIC FUNCTION OF LIVER
Galactose Tolerance Test
Procedure
Clinical Interpretations
TEST-BASED SYNTHESIS OF PLASMA PROTEINS
Clinical Interpretations
TESTS BASED ON DETOXICATION ACTION OF LIVER
Clinical Interpretations
TESTS BASED ON EXCRETORY FUNCTION OF LIVER
Clinical Interpretations
TESTS BASED ON THE SYNTHESIS OF CLOTTING FACTORS BY LIVER
ENZYME ASSAYS
Serum Transaminases
ALKALINE PHOSPHATASES—3–13 KÅ UNITS/100 ML (23–92 IU/L)
VIVA VOCE
CHAPTER 19:
Renal Function Tests
TEST FOR ASSESSMENT OF GLOMERULAR FUNCTION
Clearance Tests
Salient Features for Substance Suitable for Measuring GFR
Normal Value
Procedure
Clinical Interpretations
TESTS FOR MEASURING RENAL PLASMA FLOW
Para-amino Hippurate Test
Procedure
Clinical Interpretations
TESTS FOR TUBULAR FUNCTION
Test Procedure
Phenolsulfonphthalein Test
Test Procedure
Normal Response
Abnormal Response
Kidney Imaging
Kidney Biopsy
VIVA VOCE
CHAPTER 20:
Thyroid Function Tests
HORMONE CONCENTRATION
BINDING PROTEINS
THYROTROPIN RELEASING HORMONE RESPONSE TEST
SERUM CHOLESTEROL
THYROID AUTOANTIBODIES
THYROID SCAN
VIVA VOCE
CHAPTER 21:
Parathyroid Function Tests
PTH LEVEL
SERUM CALCIUM LEVEL
Principle
SERUM PHOSPHOROUS
Principle
VITAMIN D
BONE SCAN
VIVA VOCE
SECTION 6: DEMONSTRATIONS
CHAPTER 22:
Analysis of Saliva
COMPOSITION OF SALIVA
ANALYSIS OF SALIVA
Procedure
Collection of Saliva
Tests for Organic Constituents
Principle
Procedure
INORGANIC CONSTITUENTS ANALYSIS
VIVA VOCE
CHAPTER 23:
Analysis of Food–Stuffs
MILK
Composition
Humanization of Cow’s Milk
Pasteurization and Sterilization
ANALYSIS OF MILK
Physical Examination of Milk
Chemical Examination of Milk
VIVA VOCE
CHAPTER 24:
Biochemical Techniques
CHROMATOGRAPHY
TYPES OF CHROMATOGRAPHY
Partition Chromatography – (Martin and Synge 1941)
Adsorption Chromatography
Ion-exchange Chromatography
Gel Filteration Chromatography
Affinity Chromatography
High Pressure Liquid Chromatography
ELECTROPHORESIS
Factor Affecting Electrophoresis
Support Media for Electrophoresis
Quantification of Protein Fraction
Clinical Significance
VIVA VOCE
CHAPTER 25:
Titratable Acidity of Urine
DETERMINATION OF TITRATABLE ACIDITY
Calculation
Normal Values and Clinical Importance
VIVA VOCE
CHAPTER 26:
Nonprotein Nitrogen
PRINCIPLE
REAGENT
Nessler’s Reagent
Procedure
Normal Values and its Significance
VIVA VOCE
CHAPTER 27:
Recent Advances in Clinical Biochemistry
SPECTROSCOPY
Nuclear Magnetic Resonance/NMR Spectroscopy
Applications
Electron Spin Resonance/ESR Spectroscopy
Applications
Luminescence (Flow chart 27.1)
Applications
Light Scattering Techniques
Choice Between Nephlometry and Turbidimetry
Applications
X-ray Spectroscopy
GENE IDENTIFICATION TECHNIQUES
Southern Blotting
DNA Microarrays
GENE MULTIPLICATION TECHNIQUES
r-DNA Technology
Polymerase Chain Reaction
Applications
GENE STORAGE TECHNIQUES
CHAPTER 28:
Preparation of Various Reagents
BENEDICT’S QUALITATIVE SOLUTION
REAGENTS FOR QUANTITATIVE ANALYSIS
Creatinine Stock Standard (1 mg/ml)
Proteins
Sodium Hydroxide (0.5 N)
Sodium Phosphotungstate (for HDLc)
Sugar
Urea
Uric Acid
BCG Reagent
Biuret Reagent (Stock)
Biuret Reagent (Working)
Tungstic Acid (Diluted)
CHAPTER 29:
Normal Reference Values of Various Biochemical Parameters
INDEX
TOC
Index
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