Lab Flashcards
What method was used in the lab to measure total protein?
Biuret method
What does the biuret method consist of?
Copper sulfate reacting with peptide bonds with increasing intensity to their concentration. Colored chelate forms with amine nitrogen of the peptide bond with an absorbance at 540nm.
What type of amino acids react with the biuret method?
Tripeptides and polypeptides. Dipeptides or single amino acids do not.
What is intensity of color in biuret method results proportional to?
Number of peptide bonds and therefore protein concentration
Identify the other methods for assessing total protein.
Kjehldahl
Refractometry
UV absorption
What method is used for detecting albumin?
Dye binding technique
How does the dye binding technique work?
In dye binding technique, anionic dye binds to albumin at a pH of 4.2, pH at which albumin is cationic. BCG binds specifically to albumin although it will bind other plasma proteins over time. BCG binds to protonated amine groups on the albumin molecule and the complex forms a color at 550-630 nm
What type of specimens are used for total protein albumin measurements?
Serum
Heparinized plasma
Body fluids/urine
What is the reference interval for total protein?
6.5-8.3 g/dL
What is the reference interval for albumin?
3.5-5.5 g/dL
What is automation?
The mechanization of chemical analyses done in order to minimize manual manipulation
List the steps in the automated process
Separation/mixing Sample/reagent handling Incubation Reaction and reaction analysis Derivation/interpretation of results Result reporting
What are the advantages of automation?
Time saving
Removes human performance variability
Reduces error
Saves money
What are the disadvantages of automation?
High initial/startup cost
Discontinued or updated products and reagents
Special training required
What are discrete analyzers?
Analyzers in which samples and reagents are mixed and react in individual cucette or slide
What are continuous flow analyzers?
Analyzers in which sample is injected into a reagent stream and reactions take place in tubing