| | 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.comSpecific 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 |
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