| | 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
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