Final Practical – Flashcards
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| Magnesium and Calcium Assays Body system |
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| Bone |
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| BUN/Creatinine Assays Body system |
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| Kidney |
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| Bilirubin Assay Body system |
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| Liver |
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| Amylase Assay Body system |
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| Pancreas |
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| When working with Beer’s law, what is the nanometer used to measure? |
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| The nanometer is used to measure the wavelength of light of absorbance for the assay. |
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| Bilirubin Principle |
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| Classic diazo reaction of Ehrilich to determine bilirubin level in serum |
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| Bilirubin reagents |
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| diazotized sulfanilic acid (p-benzenediazonium) which couples with direct and indirect bilirubin to produce a color reaction |
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| Bilirubin reactions |
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| Sulfanilic acid in dilute HCl + Sodium Nitrite --> nitrous acid Nitrous acid is basis for diazotized sulfanilic acid (p-benzenediazonium sulfonate) Direct Bilirubin + diazotized sulfanilic acid at alkaline pH -> Blue Indirect Bilirubin + diazotized sulfanilic acid + accelerating agent --> Blue |
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| Creatinine principle |
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| Jaffe reaction, red-orange color forms when metabolite is treated with alkaline picrate. |
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| Creatinine reagents |
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| Picric acid |
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| Creatinine reaction |
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| Creatinine + Picric acid --> Creatinine-Picrate complex (red-orange) |
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| Total protein principle |
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| : Biuret method depends on the presence of peptide bonds, found in all proteins |
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| Total protein reagents |
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| Cupric acid |
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| Total protein reaction |
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| Cupric ions (CU2+) in alkaline solution + peptide bonds --> colored complex |
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| Amylase principle |
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| Wallenfels modification, using PNPG7 with terminal glucose blocked to reduce spontaneous degradation by reagents |
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| Amylase reagents |
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| Glucosidase, Glycoamylase, and p-Nitrophenyl D-maltoheptaosde (PNPG7) |
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| Amylase reaction |
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| PNPG7 –Amylase> PNPG3 PNPG3—glucoamylase> PNPG1 PNPG1 –glucosidase> glucose + p-nitrophenol (yellow) |
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| Calcium principle |
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| O-cresolphthalein complexone to complex with calcium to form an intense chromophore |
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| Calcium reaction |
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| CPC |
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| Calcium reagents |
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| CPC + calcium in alkaline solution --> chromophore |
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| HDL principle |
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| Quantitative determination of cholesterol concentration in HDL fraction of serum or plasma |
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| HDL reagents |
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| Magnesium chloride/dextran sulfate, enzymes |
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| HDL reactions |
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| HDL cholesterol + enzymes -->color |
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| Total cholesterol principle |
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| enzymatic methods |
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| Total cholesterol reagents |
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| cholesterol esterase, cholesterol oxidase, peroxidase |
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| Total cholesterol reaction |
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| Cholesterol Esters + H2O –cholesterol esterase> Cholesterol + Fatty Acids Cholesterol + O2 –Cholesterol oxidase> Cholest-4-en-3-one + H2O2 H2O2 + 4-Aminophenazone + phenol –peroxidase> Quinoneimine Dye + 2H2O |
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| Forms of calcium |
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| There are three forms of calcium (1) free (ionized) 50% (2) bound to plasma proteins 40% and (3) complexed with small anions 10%. |
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| The origin and breakdown of bilirubin including the enzyme involved |
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| When macrophages remove and phagocytized abnormal red cells, the heme ring is opened and heme is metabolized into biliviridin by heme oxygenase. Biliviridin is reduced by bilirubin reductase to bilirubin. Bilirubin leaves the macrophage and enters the blood stream where it is bound to albumin (indirect bilirubin). Bilirubin in conjugated to glucaronic acid in the hepatocytes by UDP glucaronic transferase and then enters the plasma in small amounts (direct bilirubin). |
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| The regulation of calcium metabolism |
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| Calcium is regulated by parathyroid hormone (PTH) which decreases excretion, calcitonin (CT) which increases excretion and Vitamin D (1, 25 (OH) 2D) which decreases excretion. High calcium levels are absorbed into the bone, and low calcium levels activate PTH which activates conversion of vitamin D into active form. Calcitonin is produced in response to high calcium levels. |
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| The regulation of phosphorus metabolism |
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| Phosphorus is regulated by PTH which decreases resorption in the proximal tubule and increases 1,25 D which stimulated intestinal absorption of phosphate. |
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| Creatinine clearance us used to measure GFR |
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| CrCl= (Urine Volume (ml/min)x UCr)/PCr |
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| Albumin |
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| 47% |
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| Alpha1-globulin |
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| 4% |
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| Alpha2- globulin |
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| 9% |
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| Beta-globulin |
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| 11% |
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| Gamma-globulin |
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| 28% |
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| Relationship between the net charge of the proteins in a sample and their electrophorectic mobility of protein fractions. |
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| Rate of electrophorectic mobility is directly proportional to net charge of the proteins, if the protein is very negative it will move faster toward the positive side. Serum proteins carry a negative charge and move toward the anode (+). |
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| Spectrophotometry |
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| The measurement of the intensity of light at selected wavelengths |
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| Chromatography |
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| A physical method of separation in which the components to be separated are distributed between two phases, one of which is stationary whereas the other moves in a definite direction. |
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| Densitometry |
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| An instrumental method for measuring the absorbance, reflectance, or fluorescence of each separated fraction on an electrophorectic strip as it is moved past a measuring optical system. |
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| Nephelometry |
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| A technique that uses a nephelometer to measure the number and size of particles in a suspension; a detector is placed at an angle to the incident light beam to measure the intensity of the light that is scattered by the particles. |
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| Amperometric measurement |
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| an electrochemical process where current is measures at a fixed potential difference between the working and reference electrodes in an electrochemical cell |
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| Acute Cirrhosis |
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| decreased albumin, decreased alpha 2, beta Gamma Bridge |
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| Chronic Cirrhosis |
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| decreased albumin, decreased alpha 2, decreased beta |
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| Acute Inflammation |
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| decreased or normal albumin, increased alpha 1, increased alpha 2, increased beta, and decreased or normal Gamma |
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| Nephrotic syndrome |
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| decreased albumin, increased alpha 2, increased beta |
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| Monoclonal Gammopathy |
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| decreased albumin, really increased gamma |
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| Polyclonal Gammopathy |
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| decreased albumin, increased gamma |
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| What effect would an air bubble have on a blood gas measurement |
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| An air bubble would be air exposure which would cause an increase in pH and pO2 and a decrease in pCO2. |
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| Henderson-Hasselbalch equation |
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| pH=pKa+log?(( [A-])/([HA])?) |
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| Henderson-Hasselbalch equation for blood gases |
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| pH=pKa+log( (HCO3-)/(0.031 x pCO2)) |
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| Specimen collection for blood gas determination |
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| Whole blood mixed well in syringe containing heparin which is usually collected from hand or arterial line. Venous blood is okay if O2 assessment is not essential. In order to maintain anaerobic conditions air bubbles must be expelled, specimen must be tested with 30 minutes because room air and WBCs will utilize oxygen content. |
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| Chylomicrons |
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| 3% |
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| VLDL |
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| 12% |
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| IDL |
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| 29% |
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| LDL |
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| 42% |
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| HDL |
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| 17-13% |
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| Calculate a specimen that has been diluted |
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| Solution is too concentrated- dilute one part sample to one part saline (1:2) - multiply results by 2 to compensate for dilution |
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| Specimen interferences Total protein |
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| Gross hemolysis and lipemia |
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| Specimen interferences HDL cholesterol |
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| freezing increases cholesterol values |
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| Specimen interferences Total cholesterol |
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| - bilirubin > 10 mg/dL, gross hemolysis and icteric specimens, fluoride and oxalate anticoagulants |
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| Specimen interferences BUN |
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| anticoagulant with ammonium salts, ammonia contamination of glassware/water, fluorides > 5mg/dL, drugs and metabolites |
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| Specimen interferences Crea |
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| - bilirubin, drugs and substances |
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| Specimen interferences Calcium |
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| calcium contaminated glassware, high bilirubin or hemoglobin, ethanol, organic solvents |
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| Specimen interferences Magnesium |
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| EDTA, glass magnesium contamination, hemolysis |
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| Specimen interferences bilirubin |
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| gross hemolysis, drugs, turbid/lipemic serum |
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| Specimen interferences amylase |
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| drugs and substances |
