CLS review pointer guidelines EXAM 2 LAB – Flashcards

Analyte Details

Calcium

Principle of the reaction:Dye binding. We said o-Cresolphthalein. This one uses

{ Calcium+Arsenazo→^alkaline →Ca-arsenazo complex}

purple color.

Reagent components.Arsenazo III (dye) and

8-hydroxyquinoline sulfonate (Mg⁺⁺   will bind to this and not the dye, this corrects the Mg⁺⁺ interference problem)

Appropriate sample:10 microliters.

Reference ranges:8.5-10.4 adults

Interferents: Mg^++,glass tubes,lipemia.

wavelength: 600-670

Analyte Details

Magnesium

Principle of the reaction:Dye binding Xylidyl Blue

serum Mg ions react with Xylidyl Blue in an alkaline solution to produce a red complex.

Reagent components:Xylidyl Blue (dye) , EGTA,(cleates Ca.), Surfactant (deproteinates),

Appropriate sample:10 microliters

Reference ranges:1.6-3.0 mg/dL adults

Interferents: hemolysis, lipemic, icteric, a number of drugs.

wavelength: 550

Analyte Details

Bilirubin (total)

 Principle of the reaction:colormetric, kinetic, end point assay-not linked.

Sulfanillic acid reacts with sodiun nitrite to produce diazotized sulfanillic acid. Direct and indirect bilirubin couple with diazo to produce azobilirubin. In the presence of dimethyl sulfoxide (DMSO). The intensity of the color produced that is produced is directly proportional to the amount of total bilirubin concentration present in the sample.

Reagent components:Sulfanillic acid, sodium nitrite, hydrocloric acid and DMSO.

Appropriate sample: 0.05 microliter

Reference ranges:0.2-1.2 mg/dL

Interferents:a number of drugs

light labile, hemoglobin

wavelength 540-560

Analyte Details

Bilirubin (direct)

 Principle of the reaction:Sulfanillic acid reacts with sodiun nitrite to produce diazotized sulfanillic acid(diazo). Direct bilirubin couples with diazo to produce azobilirubin.The intensity of the color produced is directly proportional to the amount of direct bilirubin present in the sample.

Reagent components:Sufanillic acid in hydrocloric acid, sodium nitrite reagent.

Appropriate sample:0.10 microliters.

Reference ranges:0-0.5 mg/dL

Interferents:a number of drugs, protect from light.

Wavelength: (555) 540-560 

Analyte Details

URIC ACID

 Principle of the reaction:Enzymatic endpoint assay utilizes URICASE in a linked trinder reaction.

Uric acid+O₂+2H₂O→^uricase→Allantoin+CO₂+H₂O₂

2H₂O₂+4-Aminoantipyrine+TBHBA→POD→Chromagen+4H₂O.

Uric acid is oxidized by URICASE to allntoin and hydrogen peroxide. TBHBA + 4-aminoantipyrine+ hydrogen peroxide in the presence of preoxidase produces a colored chromogen that is measured at 520. The color intensity at 520 is proportional to the concentration of uric acid in the sample.

Reagent components:uric acid reagent TBHBA, uricase(enzyme), peroxidase (for trinder)

Appropriate sample:25 microliter

Reference ranges:2.5-7.7

Interferents:bilirubin, ascorbic acid, lipemic,

wavelength :520

Analyte Details

BUN(urea)

 Principle of the reaction:Enzymatic (urease) linked kinetic (NADH).

Urea+H₂O→^Urease→2NH₃+CO₂

NH₃+∂-Ketoglutarate+NADH+H⁺→^GD→L-glutmate +NAD⁺ +H₂O

Reagent components:NADH, Urease,Glutamate dehydrogenase, Ketogluterate

Appropriate sample:10 microliters

Reference ranges: 7-18mg/dL

Interferents:ammonia, fluoride (kills urease),some drugs.

Analyte Details

Creatinine

 Principle of the reaction:Colormetric (picrate acid/modified jaffe reaction) kinetic assay

Creatinine + Sodium picrate →^Alkaline→ Creatinine-picrate complex

Yellow-orange

Reagent components:alkaline buffer, picric acid

Appropriate sample:50 microliters

Reference ranges:0.40-1.40 mg/dL

Interferents: dyes such as bromsulfophthalein and phenolsophthalein (they bind picrate) can use zinc sample pretreatment to remove these dyes before assay.

→Analyte Details

Total Cholesterol

 Principle of the reaction:

Cholesterol Esterase→C Esters→Cholesterol+Fatty acids

Cholesterol+O₂→C. Oxidase→Cholesterol-3-one+H₂O₂

2H₂O₂+ 4-Aminoantipyrine+p-HBS→Peroxidase→Quinoneimine+2H₂O

(trinder)

Reagent components:4-Aminoantipyrine, Sodium Cholate, Cholesterol esterase,cholesterol oxidase, Horseradish Peroxidase, P-Hydroxybenzene Sulfonate, Surfactant,

Appropriate sample:10 microliters

Reference ranges:<200mg/dL

Interferents:

Wavelenght 520

Analyte Details

HDL Cholesterol

precipitation

 Principle of the reaction:When serum is combined with reagent dextran sulfate and magnesium ions precipitate the LDL and VDRL fractions, leaving the HDL fraction in solution. The HDL cholesterol is then determined using enzymatic cholesterol assays

Reagent components:HDL cholesterol Precipitating reagents, dextran sulfate, Magnesium ions, sodium.

Appropriate sample:50 microliters

Reference ranges:27-78

Interferents: Hemolysis, icteric, ascorbic acid

Analyte Details

HDL Cholesterol

Direct

 Principle of the reaction:The liquid auto HDL Cholesterol assay is a homogeneous method for directly measuring serum HDL-C levels without the need for any off-time pretreatment or centrafugation steps. The method is a two reagent format. The first reagent contains a-clyclodextran and dextran sulfate to stabilize LDL, VLDL and cholymicrons. (so they don't react with the second reagent). The second reagent contains PEG modified enzymes that selectively react with the cholesterol present in the HDL particles. Consequently, only the HDL cholesterol is subject to cholesterol measurement.

Reagent components:clyclodextran, PEG

Appropriate sample:4 microliters

;

friedwald formula

;

LDL-C=Total Cholesterol -HDL-C - Trig/5

Analyte Details

Triglycerides;

;Principle of the reaction:linked pyruvate kinase enzymatic ultra violet.Triglycerides in the sample are hydrolyzed by by lipase to glycerol and fatty acids. The glycerol is then phosphorylated by adenosine-5-triphosphate (ATP) to glucerol 3 phosphate(G3P) and adenosine-3-diphosphate in a reaction catalyzed by glycerol kinase(GK). Glycerol3 phosphate is then converted to dihydroxyacetone phosphate (DAP)and hydrogen peroxide by glycerol phosphate oxidase(GPO). The hydrogen peroxide then reacts with 4-aminoantipyrine (4-AAP) and 3-hydroxy-2-4-6 tribrombenzoic acid (TBHB) in a reaction catalyzed by peroxidase to yeald a red colored quinoneimine dye. The intensity of the color produced is directly proportional to the concentration of triglycerides in the sample when measured at 540.

Reagent components: ATP, LIPASE, TBHB, Peroxidase, surfactant, (deproteinator)

Appropriate sample:10 microliters.

Reference ranges:36-165 mg/dL

Interferents:hemolysis glycerol, bilirubin, detergents, drugs

Analyte Details

Serum;Iron;

;Principle of the reaction:Transferrin bound iron is released at acid pH and reduced from ferric to ferrous ion these ions react with ferrozine to form a violet colored complex

Reagent components: acid buffer and ferrozine

Appropriate sample: 0.5ml (500microliters)

Reference ranges: 60-150 ug/dL

Interferents:drugs testtubes, not hemoglobin

Analyte Details

Serum;Iron;(TIBC)(UIBC)

;Principle of the reaction:Transferrin bound iron is released at acid pH and reduced from ferric to ferrous ion these ions react with ferrozine to form a violet colored complex.

(TIBC) A known amount of ferrous ions are added to serum at an alkaline pH. The ferrous ions bind with transferrin. the additional unbound ferrous ions are measured using the ferrozine reaction.The difference between the amount of ferrous ions added and the unbound ions measured is the unsaturated iron binding capacity.The TIBC is equal to the serum iron concentration plus the UIBC.

Reagent components:alkaline and acid buffer and ferrozine

Appropriate sample:0.5ml (500microliters)

Reference ranges:60-150 ug/dL

Interferents:drugs testtubes, not hemoglobin

Sweat cloride

Method

pilocarpine nitrate iontrophoresis

Collection of sweat and gavimetric measurement of collected sweat.

Can cause burns, current interupts heart.

Trained staff

Monitor electrodes

monitor electrical feed

time

do not cross chest.

Arsenazo

complexes with calcium to form a purple colored complex

;

and o-Cresophthalein