1 / 22

CM3107 BIOANALYTICAL/CLINICAL ANALYSIS

CM3107 BIOANALYTICAL/CLINICAL ANALYSIS. Prof. George G. Guilbault Room 205, Robert Kane Building University College Cork, Ireland. CLINICAL BIOMEDICINE STUDY OF CHEMISTRY OF HUMAN SYSTEM -WHAT CHEMICALS ARE INVOLVED IN DISEASES - WHICH ONES AFFECT WELL BEING

merrill
Download Presentation

CM3107 BIOANALYTICAL/CLINICAL ANALYSIS

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. CM3107BIOANALYTICAL/CLINICAL ANALYSIS Prof. George G. Guilbault Room 205, Robert Kane Building University College Cork, Ireland

  2. CLINICAL BIOMEDICINE STUDY OF CHEMISTRY OF HUMAN SYSTEM -WHAT CHEMICALS ARE INVOLVED IN DISEASES - WHICH ONES AFFECT WELL BEING - WHY IMPORTANT 2/3 OF ALL ANAL WORK IS IN THIS AREA 1 OF 2 OF ALL PERSONNEL IN THIS AREA HISTORY OF CLINICAL: l. OTTO WARBURG IN 1940 – NADH ABSORBS AT 340nm 2. 1965 OVERHAUL OF CLINICAL ANALYSIS(NEXT) 3. 1970 – SKEGGS - AUTOMATION

  3. HIGH GLUCOSE = DIABETES- IMBALANCE OF METABOLISM SUCROSE  GLUCOSE  GLYCOGEN IN 1965 NATIONAL INSTITUTES OF HEALTH DID A STUDY OF THE ABILITY OF ANALYTICAL LABS TO “ GET THE CORRECT RESULT” TRUE RESULT WAS NORMAL: 90 mg/dL 30% OF LABS REPORTED HIGH(RESULT:INJECTION OF INSULIN  DEATH) 40% OF LABS REPORTED NORMAL 30% OF LABS BELOW NORMAL(RESULT:INJECTION OF SUGAR LETHAL TO DIABETIC) RESULT: BIG SHAKEUP IN LABS LEADING TO BETTER RESULTS AND A MODERNIZATION OF ANALYSIS.

  4. MOST IMPORTANT TESTS l. GLUCOSE – HIGH  DIABETES( IMBALANCE OF METABOLISM) 2. UREA/CREATININE – MALFUNCTIONING OF KIDNEY 3. CHOLESTEROL/HDL/LDL/VLDL – MYOCARDIAL INFARCT(HEART) 4. URIC ACID – GOUT 5. ALKALINE PHOSPHATASE – DAMAGE TO LIVER 6. AND OF COURSE – PHARMACEUTICAL INDUSTRY  BENEFITS BY PROVIDING NEW DRUGS FOR THESE DEFICIENCIES BIGGEST – DIABETES – NEW DRUG RECENTLY APPROVED FOR TYPE 2 DIABETES = JANUVIA

  5. PURPOSE OF CLINICAL LAB=QUAL AND QUANT ANALYSIS OF BODY FLUIDS-BLOOD,URINE,SPINAL FLUID,FECES,etc UNLIKE ALL OTHER ANALYSIS HERE WE USE LIQUID NOT MASS(TAKE FEW MICROLITERS BLOOD).USE dL AS UNIT.IN IRELAND/UK USE mM,ALL OTHERS mg/dL: Mg per liter/Molecular Weight = mM

  6. CURRENT TYPICAL VALUES IN PLASMA GLUCOSE,mg% 90 (5.0 mM) UREA,mg % 16 CREATININE,mg% 1.1 URIC ACID, mg% 4.6 TOTAL CHOLESTEROL 194 POTASSIUM,meq/liter 4.4 ALKALINE PHOSPHATASE,units 3.0 LDH, units 360

  7. ORIGINS OF SPECIES DETERMINED STAGE 1 – TESTS ARE ON ENZYMES OR SUBSTRATES COMING FROM SPECIFICALLY DAMAGED ORGANS l. ALKALINE PHOSPHATASE – DAMAGED LIVER HIGH: OBSTRUCTIVE JAUNDICE,PADGETTS DISEASE, RICKETS 2. CHOLINESTERASE – DAMAGE TO NERVE ENDINGS HIGH: NEPHROTIC SYNDROME LOW: PESTICIDE POISONING, ANEMIA, MALNUTRITION 3. LACTATE DEHYDROGENASE(LDH) MYOCARDIAL INFARCTION, LEUKEMIA 4. GLUCOSE – DIABETES(TYPES 1 AND 2) 5. UREA/CREATININE – DISEASES OF KIDNEY

  8. QUALITY CONTROL AND VALIDATION • VERY IMPORTANT IN CLINICAL LABS WHERE LIFE/DEATH DECISIONS ARE MADE • Reference: N. TIETZ-FUNDAMENTALS OF CLINICAL CHEMISTRY • MUST CALIBRATE INSTRUMENT TWICE DAILY WITH STANDARD BLOOD SAMPLE WITH KNOW VALIDATED VALUES • B. MUST USE REAL BLOOD SAMPLES: • l. POOLED SERUM • 2. FREEZE DRIED SERUM CONTROLS • a. DADE – MONITROL: NORMALS FOR ALL SUBSTRATES(eg GLUCOSE),METALS, ENZYMES • b. LEDERLE LABS – LEDERNORM(MOST TESTS)

  9. NORMAL VS ABNORMAL WHAT IS NORMAL = AVERAGE PERSON(?) -ACCEPTED AS ABOUT 80% OF POPULATION -RANGE IN WHICH 95% FIT(1 in 20 OUTSIDE) - 80 to 95% OF POPULATION ARE GREY AREA - 5% CONSIDERED ABNORMAL(See TIETZ) DADE FOR EXAMPLE CAREFULLY SCREENS DONORS FOR DISEASES, MEDICAL HISTORY. -PICKES SEVERAL HUNDRED,COLLECTS BLOOD=FORMS A POOL. FREEZE DRY BLOOD, ASSAY AND REPORT ALL VALUES(SEPARATE FOR EVERY DIFFERENT INSTRUMENT).ONLY ABOUT 5% TAKEN AS REPRESENTATIVE FOR ANALYSIS. -ABNORMAL- SPIKE WITH PURE REAGENTS.ASSAY

  10. TYPES OF ASSAYS l. EQUILIBRIUM – LET ALL REACT, MEASURE AT EQN A + B  PRODUCTS MEASURE PRODUCTS BY COLOR, FLUORESCENCE, ELECTROCHEMICAL METHOD 2. KINETICS MEASURE RATE OF REACTION AS AFFECTED BY A ABOVE AT CONSTANT B. 4 X C1 ABSORBANCE 3 X C1 2 X C1 C1 TIME

  11. ASSAY OF ENZYMES MUST BE DONE BY KINETIC(RATE ) METHOD SINCE THEY ARE CATALYSTS ADVANTAGES: THEY ARE HIGHLY SPECIFIC AND SENSITIVE BASE EQUATIONS E + S(k1/k2)  ES (k3) PRODUCTS + E Vo = Vmax[So]/Km + So where Vmax = k3[Eo] Km = k2 + k3/k1 ENZYME ACTIVITY = HOW MUCH SUBSTRATE IS CONVERTED AMBIGUITY ARISES SO USE INTERNATIONAL UNITS IN OLD SYSTEM WE HAD MANY ASSAYS WITH DIFFERENT CONDITIONS/IMPOSSIBLE TO CORRELATE THERE ARE A NUMBER OF IMPORTANT ENZYMES WE WILL LOOK AT AND ALSO ISOENZYMES.

  12. TO SYSTEMATIZE: COMMISSION ON ENZYMES IF INTERNATIONAL UNION OF BIOCHEMISTRY IN 1961 MET IN NEW YORK: RECOMMENDATIONS OF I.U.B., ELSEVIER, 1965 ONE UNIT = AMOUNT OF ENZYME THAT CATALYZES REACTION OF ONE MICROMOLE OF SUBSTRATE PER mg ENZYME PER MINUTE AT STANDARD CONDITIONS(USUALLY T=30 C,pH OPTIMUM ETC) RECOMMEND U/mL or U/L at 30 C REMEMBER: ENZYMES ARE SECONDARY INDICATORS OF CELL OR TISSUE ACTIVITY- IF UP OR DOWN= SOMETHING WRONG.BETTER = DIRECT ASSAY OF TISSUE. THIS IS DIFFICULT:IN SECOND GENERATION TESTS WE USE (l) IMMUNOCHEMISTRY,eg PSA TEST .

  13. ENZYMES AND DISEASES. ENZYME ORGAN OF DISEASE SPECIMEN Acid Phosphatase Prostate Serum Alkaline Phosphatase Liver, bone Serum/urine Amylase Pancreas Serum/urine Lipase Pancreas Serum/urine Lactate Dehydrogenase(LDH)Liver,Heart Serum/urine Aspartate transaminase(GOT) Liver,Heart Serum/urine Alanine transaminase(GPT) Liver,Heart Serum/urine Creatine Hinase(CK) Heart,Muscle,Brain Serum Aldolase Copper transport disease(Wilson) Serum

  14. USE OF PROFILING SEVERAL TESTS USED TO INDICATE A SPECIFIC FUNCTIONING OF BODY WHY-COVER ALL POSSIBILITIES(AVOID LAWSUITS) l. LIVER : LDH, GOT,GPT,ALK PHOSPATASE,GAMMA GLUTAMYL TRANSPEPTIDASE(GGTP) 2. KIDNEY : UREA, CREATININE, GGTP 3.PANCREAS : LIPASE, AMYLASE,LEUCINE AMINOPEPTIDASE 4. MUSCLE: CK, LDH 5. BONE: ALKALINE PHOSPHATASE –TARTRATE INHIBITED(ISOENZYME 1)OR UREA INHIBITED(ISO 2) 6. CANCER : ALK AND ACID PHOSPHATASE,LDH,ASPARAGINASE,LAP

  15. ISOENZYMES=MAJOR IMPROVEMENT in ASSAYS WHAT ARE THEY:SOME ENZYMES FROM AN INDIVIDUAL ORGANISM CAN EXIST IS SEVERAL DIFFERENT FORMS=ISOENZYMES DIFFERENCES- l. CHARGE = CAN BE SEPARATED BY ELECTROPHORESIS 2. STABILITY TO HEAT DENATURATION 3. REACTION TO CHEMICAL INHIBITORS(Alk Phosphatase) 4. AFFINITY FOR SUBSTRATES OR COENZYMES WHICH ONES ARE COMMON: LDH,CK,ALK AND ACID PHOSPHATASE(PERHAPS ALL BUT NOT GLUCOSE OXIDASE)

  16. ANALYSIS OF ENZYMES • WHICH ARE MOST IMPORTANT: • ALKALINE PHOSPHATASE –SIGNIFICANCE: • LIVER DISEASE INDICATOR, OBSTRUCTIVE JAUNDICE,PADGETTS AND OTHER LIVER DISEASE INDICATORS • ASSAY: PHENYLPHOSPHATE  PHENOL + Pi • PHENOL + DIAZOREAGENT  RED COLOR • ACTIVATORS – Mg(II) AND Mn(II) • OR p-NITROPHENYLPHOSPHATE  p-NITROPHENOL(410nm) • OR FLUOROMETRIC USING NAPTHOL ASBI PHOSPHATE GREEN FLUORESCENCE

More Related