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CLINICAL BIOCHEMISTRY ASSAYS GLUCOSE PROFILE FBS/RBS (FPG) GTT 2HR PP HbA1c URINE sugar/ketones ALT ALP GGT TOTAL BILIRUBIN Direct Indirect TOTAL PROTEIN Albumin Globulins BUE URIC ACID CREAT CLEAR. Mg2+ CREAT Na+ K+ ClLIVER FUNCTION TEST(LFT) AST KIDNEY FUNCTION TEST(KFT) BUN TOTAL CHOLES TRIG HDL-C LDL-C VLDL LDH CK-MB MYOGLO TROPONIN AST TOTAL IRON TIBC Ferritin ALB Ca2+ PO42ALP CREAT LIPID PROFILE CARDIAC PROFILE IRON STUDIES BONE PROFILE
HORMONAL ASSAYS THYROID TEST T4 T3 TSH FN FERTILITY HORMONES FSH LH PRL PRG E2 TUMOUR MARKERS Total PSA B-HCG CEA AFP CA 125 OTHER HORMONES INSULIN CORTISOL OTHER CHEMISTRIES AMYLASE
LAMBERT-BEERS LAW: The [substance] is directly proportional to the amount of light absorbed or inversely proportional to the logarithm of the transmitted light. C A log 100 %T A=abC Where A= absorptivity, b=length of light path, C= [ ] a and b are constants. C1=A1, C2=A2, C1 = A1 C2 A2 [ Ctest= Cstd A test] Astd Beers law is the basis of all the spectrophotometric measurements. Advantages of spectrophotometric measurements: 1. High sensitivity 2
Urinalysis is an indispensable part of chemical pathology. It is used to uncover dx anywhere in the urinary tract. TESTS ON URINE COMMONLY INCLUDES A) URINE COLOUR AND APPEARANCE (MACROSCOPY): Its colour is determined by its concentration, the presence of drugs, exogenous and endogenous compounds and its pH. Colourless urine---normal or 20 to diuretic use, high fluid intake,DIorDM. Cloudy/Hazy------phosphates, pyuria,or bacteruria Yellow to orange----- presence of bile, riboflavin , bilirubin, urobilinogen Red----------------presence of haemoglobin, Amber B)CHEMICAL COMPONENTS TEST Specific gravity pH Protein Glucose Ketones Bilirubin REFERENCE 1.003-1.029 4.5-7.8 Negative Negative Negative Negative
C )MICROSCOPY COMPONENTS: Microscopy components consists of epithelial cells, pus cells, RBCs,WBCs,casts. These are examined microscopically.
PRACTICAL II
An essential metabolic fuel and its concentration in the blood are normally tightly controlled
by the action of insulin and the counter regulatory hormones like Glucagon, cortisol, catecholamine, and growth hormone. [Glu] rises after meals and stabilizes at lower levels on fasting. Syno: blood sugar, FBS, FBG sugar. Specimen: plasma: Note: whole blood taking in fluoride oxalate vacutainer. Patient care: Pt should be fasting for 8hours-12hours prior to the test. Source of error: blood not in fluoride oxalate tube. Glycolysis by red cells will consume glu and may lower concentrate in vitro in absence of fluoride. BLOOD & PLASMA GLUCOSE Plasma [glu] is 10-15% higher than whole blood [glu] since red cells contain less water per unit volume than plasma. The discrepancy can be greater them this if [glu] is changing rapidly because glu will not have reached equilibration across the red cell membrane. Therefore plasma yields more reliable results.
Types of glucose test: RBS, 1hr pp, 2hr pp. 2hr Postprandial: glucose levels 2hrs after meal or after measured glucose level. It is used extensively to establish the diagnosis of DM Ref Ranges: FBS : 3.6-6.4 mmol/L RBS :3.3-7.0mmol/L (Dependant on time & meal content)
Take samples of Times 0. FBS 1. 1 hr pp 2. hr pp 3. 3hr pp. with urine samples H2O GLUCOSE LOAD: g in 300 ml H2O RESULTS OF OGTT Vs TIME 5
TIME (Min) 0
FASTING BLOOD SUGAR
URINE SUGAR
60 120 180 METHODOLOGY: Glucose oxidase method or Hexokinase method. Principle of method: glucose oxidase (GOD) catalyzes the oxidation of glucose to give hydrogen peroxide (H2O2) and gluconic acid. In the presence of enzyme peroxidase (POD), the hydrogen peroxide is broken down and the oxygen released reacts with 4-aminophenazone (4-aminoantipyrine) and phenol to give a pink colour. The absorbance of the colour produced is measured in a spectrophotometer at 520nm. Glu+O2 + H2O Glucose + phenol + 4Aminophenazone PROCEDURE STD Glucose Rgt STD TEST 10L 10L 10L oxidase 1 ml STD 1ml TEST 1ml BLK 1ml
G Oxidase
Mix well and incubate at room temperature for 10mins or in water bath for 5mins and read absorbance at 520nm against reagent blank. Calculation; [test] = [std] Abstd CAUSES OF ABNORMAL RESULTS HIGH 1. DM a. Type I (IDDM) b. Type II (NIDDM) c. Impaired glucose tolerance d. Gestational diabetes 2. Pancreatitis 3. Endocrine disorders eg. thyrotoxicosis 4. Drugs eg. steroids, oral contraceptives 5. Chronic renal failure 6. Stress 7. I.V. glucose infusion 8. Postprandial 4. Extra-pancreatic neoplasm 5. Serum liver dysfunction 6. Ethanol ingestion 7. Drugs eg. sulfourea, salicylates, insulin 2. Adrenal cortical insufficiency acromegaly, 3. Hypopituitarism LOW 1. Insulinoma Abtest mmol/l