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Jordan University Faculty of Science, Department of Biological Sciences
B0304323 (4-Credit-Hr): Clinical Chemistry
Lecturer: Dr. Ghaleb M. Abuereish, Ph.D. (Professor). Office: Biology 208.
Course Contents & Grading System. February 7, 2010.
Primary Reference Textbook: Clinical Chemistry: Principles, Procedures, Correlations, 5th edition (2005) by
Bishop ML, Fody EP, Schoeff LE. Lippincott Williams & Wilkins Publisher.
http://www.LWW.com
Specific References are listed at the end of each topic in the textbook.
Chapter                                    Topic                                                                                    Page
1 Basic principles and practice of clinical chemistry:     1-33
 Units of measure. Reagents. Chemicals. Reference materials. Water
 specifications. Solution properties. Concentration. pH and buffers. Laboratory
 mathematics and calculations: significant figures and logarithms, percent
 solution, molarity, normality, specific gravity, conversions, dilutions, water of
 hydration, graphing Beer’s law, enzyme calculations. Specimen considerations.
8 Aminoacidopathies:         180
  Phenylketonuria      
  Alkaptonuria
  Maple syrup urine disease
  Homocystinuria
  Cystinuria
 Plasma Proteins: Functions & Clinical Significance     193
  Prealbumin (transthyretin)
  Albummin
  Globulins: α1-antitrypsin, α1-fetoprotein, Gc-binding protein,
haptoglobin, ceruloplasmin, transferrin, hemopexin, fibrinogen,
C-reactive protein, & Immunoglobulins (Igs).
 Miscellaneous proteins:       
  Troponin         200
  Methods of analysis        203
  Total protein methods
  Biuret, dye binding, ultraviolet absorption.
Electrophoresis: Normal vs abnormal patterns    207
9 Nonprotein nitrogen compounds       219
 Urea, Creatinine/Creatine, Uric acid, & Ammonia:
  Disease correlation
  Analytical methods
10 Enzymes          237
  Measurement of enzyme activity      242
  Calculation of enzyme activity      243
 Enzymes of clinical significance:       243
  Creatine kinase (CK), Lactic acid dehydrogenase (LDH),
  Aspartate minotransferase (AST), Alanine aminotransferase (ALT),
  Alkaline phosphatase (ALP), Acid phosphatase (ACP),
  γ-Glutamyltransfease (γ-GT), Amylase (AMS), Lipase (LPS),
  & Glucose- 6 – phosphate dehydrogenase (G-6-PD).

11 Carbohydrates          262
 Regulation of carbohydrates metabolism      267
 Hyperglycemia: Diabetes mellitus       268
  Classification of diabetes
  Criteria for the diagnosis of diabetes mellitus
 Hypoglycemia          273
 Methods of glucose measurements       275
 Glycosylated hemoglobin: HbA1c (A1c)      277
12 Lipids & Lipoproteins         282
 General lipoprotein structure & characteristics of the major human lipoproteins:
 Chylomicrons. Very-Low-Density-Lipoproteins. Low-Density-Lipoproteins.
 Lipoprotein (a). High-Density-Lipoproteins.
 Characteristics of the major human apolipoproteins.
 Lipoprotein physiology & metabolism
 Hyperlipoproteinemia. Hypercholesterolemia. Hypertriglyceridemia:
 Coronary heart disease. Arteriosclerosis. Atherosclerosis.
 Lipid & Lipoprotein Analysis: Measurement      298
13 Electrolytes          314
 Sodium (Na+). Potassium (K+). Chloride (Cl-). Bicarbonate (HCO3-).
 Magnesium (Mg++). Calcium (Ca++). Phosphate.
14 Blood gases, pH, & Buffer systems       343
 Regulation of Acid-Base Balance: Lungs & Kidneys
 Acidosis. Alkalosis
 Blood gas analysis: pH. pCO2. pO2.
17 Introduction to Hormones & Pituitary Function         399-409
22 Liver Function  Tests         475
  Excretory & Secretory Function: Bilirubin
 Analysis
 Enzyme Tests in Liver Disease: ALP. AST. ALT. 5’-Nucleotidase.
 γ-GT. Leucine aminopeptidase. LDH.
 Hepatitis
23 Cardiac Function:          496
 Laboratory Diagnosis of Acute Myocardial Infarction    505
 Enzymes: CK, CK-myoglobin (CK-MB), Troponin T,  Troponin I, & MB.
 Markers of Inflammation & Coagulation Disorders:     507
 Hs-CRP, Fibrinogen, D-Dimer.
 Markers of Congestive Heart Failure: Brain-type, or B natriuretic peptide (BNP).
 Other Markers: Glycogen phosphorylase BB, Heart Fatty Acid-Binding Protein,
 Carbonic anhydrase III, Ischemic-modified albumin, & Homocysteine.
24 Renal Function         517
 Kidney Function Test: Analytic Procedures
 Clearance measurements, Creatinine, Creatinine clearance & GFR, Urea.
 Urine Electrophoresis: β2-Microglobulin, Myoglobin, Microalbumin, Cystatin C.
 Urinalysis
 Glomerular Diseases         529
25 Pancreatic Function: Tests        541
26 Gastrointestinal Function: Tests of Intestinal function    548
  Lactose tolerance test, D-xylose absorption test, Fecal fat analysis
30 Circulating Tumor Markers: Basic Concepts & Clinical Applications
 Clinical Utilities of Tumor Markers:
            α-Fetoprotein (AFP), Prostate-Specific Antigen (PSA). 
 Types of tumor Markers: Enzymes, Serum Proteins, & Hormones.
CarcinoEmbryonic Proteins (CEA)        610
 Monoclonal Defined tumor Markers: Immunoassays Monoclonal kit  611
  CA 125     Ovarian Carcinoma
  Hybri-BREScan  (CA 495 or CA 15-3) Breast Cancer
  Hybri-CMark (CA 195 or CA 19-9)  Pancreatic Carcinoma
  CA 72-4     Gastric Carcinoma
  Free PSA Differentiate between BPH & Prostatic Carcinoma
 Cell-Specific Tumor Markers:       611
Clinical Laboratory
Experiment    Experiment
 1 Check in & Introduction to lab techniques, blood sampling  preparation of reagents.
 2 Measurement of total protein & albumin in serum. Serum Protein electrophoresis.
 3 Measurement of urea & creatinine in serum.
 4  Measurement of bilirubin in serum
 5 Assay of LDH in serum. Assay of ALP & ACP activities in serum
 6         Assay of ALT & AST activities in serum
 7 Measurement of glucose & uric acid in serum.
 8 Measurement of cholesterol in serum + HDL + Triglyceride
 9 Assay of Amylase & Lipase in serum, and assay of HbA1c.
10 Measurement of Na & K in serum by flame photometry.
11 Qualitative measurements of   urinary glucose, proteins, bilirubin, urobilinogen, nitrite,
            ascorbic acid, & pH.
12 Measurement of Ca & Pi in serum
13 Measurement of Fe & total iron binding capacity (TIBC) in serum.
 Final lab Exam
Grading System:
First Exam = 20% (15% for written exam plus 5 % lab reports). Midterm Exam = 30% (22.5 % for the lectures & 7.5% for the written midterm lab). Final Exam = 50% (37.5% for the lectures & 12.5% for the final written lab).
Bonus: 10% for attendance, discussion participation, and lab practical exam.
2008, 2009, 2010
  
Jordan University Faculty of Science, Department of Biological Sciences
B0304213 (2-Credit-Hr): Laboratory Instruments & Methods of Measurements
Lecturer: Dr. Ghaleb M. Abuereish, Ph.D. (Professor). Office: Biology 208.
Course Contents & Grading System: February 7, 2010.
Primary Reference Textbooks: (1) Clinical Chemistry: Techniques & Instrumentation: A practical Course
by Varcoe, J. S. (2001), World Scientific Co., NJ. 
Specific References are listed at the end of each topic in the textbook.
Week No.          Topic                                                                                                     
1.  Introduction
2. Spectrophotometry:
   - Photometric analysis
     Electromagnetic spectrum of interest, pages 1-1 to 1-2.
     Wavelengthmax, λmax, and ε, pages 1-4
   - Laws of radiant energy absorption, pages 1-5 to 1-8
   - Color & Colorimeters, pages 1-9 to 1-34
              - Spectrophotometer instrumentation: energy sources, spectral isolation
                   cuvettes, detectors & readout systems.
3.   - Single-beam spectrophotometers
   - Double-beam spectrophotometer
   - Instrument performance, light wavelength, & spectral band width.
4.           - Turbidimetry & Nephelometry   
 Exercises pages 1-35 to 1-62: 1-1,2,3,4,5,6,7,8,9,11,12,13,14. 
5. Fluorimetry: pages 2-1 to 2-10.
             - Fluorescence
             - Factors affecting fluorescence intensity: concentration, sources, buffers,   
                 pH & ionization, temperature, photodecomposition & quenching.
 - Fluorimeter instrumentation: light source, sample container, detectors & its sensitivity.
 - Clinical application of spectorfluorimeters.
 Exercises pages 2-11 to 2-30: 2-1,2,3,4,5,6,7,8,9,10.
6. Atomic Emission & Absorption: pages 4-1 to 4-14 
 - Flame emission photometry:
  - Principles of flame photometry
  - Components of flame photometer: nebulizer, burner, monochromator, light detector, & 
                  readout device.
7.  - Atomic absorption spectrophotometry:
    Flame atomic absorption. Interference & matrix effect.
 - Chemical interference: incomplete  dissociation & ionization of compounds  
            - Electrothermal atomization (flameless atomic absorption).
 Exercises pages 4-15 to 4-33: 4-1,2,4,5.
8. Ion selective electrodes: pages 5-1 to 5-8.
 - Principle of ion selective electrodes
   Reference electrodes. pH, proton (H+) electrode. Sodium, Na+, electrode. Potassium,
              K+, electrode. Calcium, Ca, electrode.
 Exercise 5-1.
9. Gas electrodes: pages 6-1 to 6-5.
 - Oxygen, O2, electrode. Carbon dioxide, CO2, electrode.
            - Blood gas analyzer
10.  Chromatography: pages 7-1 to 7-4
 - Adsorption chromatography
 - Partition chromatography
 - Gel permeation chromatography
 - Ion exchange chromatography
 - Qualitative & quantitative analysis
 Exercises: 7-2,3,5.          
11. Thin layer chromatography: pages 8-1 to 8-4
 - Extraction techniques
 - Thin layer chromatography
 Exercise 8-1.
12.  Gas chromatography: pages 9-1 to 9-22.
 - Components of gas chromatography:
   Carrier gas. Flow control. Sample injection systems. Thermostatically controlled oven.
   Columns & their packing. Selection of stationary phase.
              Detectors: thermal conductivity detectors, flame ionization detectors, nitrogen – phosphorus     
              detectors, electron capture detectors. Recorder & computing integrator
 - Qualitative & quantitative analysis
13.  High performance liquid chromatography: pages 10-1 to 10-27
 - Introduction.
 - Equipment components:
    Mobile phase reservoir. Pump.Gradient elution. Guard column. Analytical columns
14. Electrophoresis: pages 11-1 to 11-18.
 - Principle of electrophoresis
 - Moving boundary electrophoresis
 - Zonal electrophoresis
 - Electrophoresis support media:
 -  Cellulose acetate. Agarose gel. Polyacrylamide gel
 - Isoelectric focusing electrophoresis
 - Two dimensional electrophoresis
 - Capillary electrophoresis
 Exercise 11-2,3,4,6
15. Immunological & Radioisotope techniques: pages 13-1 to 13-40
 Immunological techniques:
   Double immunodiffusion. Single immunodiffusion. Crossed immunoelectrophoresis
   Electroimmunoassay. Immunoassay.
 Radioimmunoassay (RIA):
   Immunoradimetric assay. Enzyme Linked Immunosorbent Assay.
              Enzyme multiplied immunoassay techniques. Immunoprobes & immunosensors.
Grading system: Firs exam = 20%. Midterm = 30%. Final = 50%.
Office hours will be announced.
Lab exercises as in Primary Reference Textbooks: (1) Clinical Chemistry: Techniques & Instrumentation: A practical Course by Varcoe, J. S. (2001), World Scientific Co., NJ. 
Lab No.  Exercise                      page
1    1-1   Photometric accuracy and linearity      1-36
2    1-7   The absorption spectrum of benzene      1-42
3    1-8   The absorption spectrum of NAD and NADH     1-43
4    1-11 The determination of serum calcium      1-47
5    1-13 The determination of plasma salicylate     1-55
6    1-14 The determination of plasma paracetamol     1-58
7    2-1   The effect of concentration on fluorescence intensity    2-11
8    2-3   The effect of pH on fluorescence intensity     2-13
9    2-6   The emission spectrum of NADH       2-17
10    4-1    The determination of Na and K by flame photometry    4-15
11           4-3    The determination of Ca & Mg by flame atomic absorption (Geology)  4-19
12    5-1   Determination of the pH of urine       5-11
13    6-3   Blood gas determinations (University Hospital)     6-11
14           7-5   The determination of blood hemoglobin A1c
                        using ion exchange chromatography       7-21
15    Gas chromatography, HPLC, & Mass spectorometry (Chemistry Department) 9-1
16    11-3  Agarose gel electrophoresis of serum or plasma proteins   11-29
2008, 2010
  
 
2008, 2010