CC2 Lab: Phosphorus and Gamma Glutamyl Transferase

gamma glutamyl transpeptidase

gamma glutamyl transferase also called as?

liver, bile duct

GGT is an enzyme present in where?

hepatobiliary disease

GGT is the most sensitive indicator for?

high negative predictivity value

because of this characteristic of this test for these disease, the measurement of GGT is widely used to rule out a hepatic and biliary origin.

rule out hepatic and biliary origin

measurement of GGT is widely used to?

aspartate, alanine aminotransferase, cholinesterase

what other enzymes are used with GGT as a valuable tool for the differential diagnosis in liver diseases?

kinetic photometric test according to Szasz and Persjin

what is the method of GGT?

the test has also been standardized to the method according to IFCC. Results according to the IFCC are obtained using a special factor or in case a calibrator is used, by use of the calibrator value given for the IFCC method

explain the whole GGT method

glutamic acid

GGT catalyzes the transfer of what? to acceptors like glycylglycine in this case

5-amino-2-nitrobenzoate, 405nm

the process of GGT and glycylglycine, releases what product? which can be measured at what wavelength?

activity of Gamma-GT

the increase in absorbance at 405 wavelength is directly related to?

L-gamma-glutamyl-3-carboxy-4-nitroanilide

what is added with glycylglycine for GGT to make 5-amino-2-nitrobenzoate?

TRIS (pH 8.28) 135 mmol/L, glycylglycine 135 mmol/L

what are the components and concentration of R1?

L-gamma-glutamyl-3-carboxy-4-nitroanilide (pH 6) 22 mmol/L

what are the components and concentration of R2?

expiry date, 2-8C, free of contamination, do not freeze

reagents are stable up to? and what are the conditions to make it stable?

lights all the time

reagent 2 must be protected from?

sodium azide (0.095%)

what does the substrate and buffer contains?

never swallow, avoid skin contact and mucous membrane

what shouldn’t you do with sodium azide?

ready to use

in substrate start the reagents are?

mix 4 parts R1, 1 part R2

how to prepare for for a sample start? (like 20 ml of R1 for a 5 ml R2)

monoreagent

sample start is also called a?

serum, heparin plasma

what specimens are acceptable for GGT?

4 weeks at 2-8C, 5 days at 15-25C

enumerate the stability of GGT specimen

1421, 1606

enumerate the factors of substrate start at 405nm in Szasz and IFCC

1158, 1309

enumerate the factors of sample start at 405nm in Szasz and IFCC

GGT (U/L) x 0.0167 = GGT (ukat/L)

what is the conversion formula of GGT?

ascorbic acid up to 30 mg/dL, bilirubin up to 40 mg/dL, hemoglobin up to 400 mg/dL, lipemia up to 2,000 mg/dL triglycerides

enumerate the substances and their concentrations that were observed to have no interferences

2 U/L

what is the lower limit detection of the GGT?

women <32 u/L (<0.53 ukat/L), men <49 u/L (0.82 ukat/L)

enumerate the reference values of GGT according to Szasz

female <38 u/L, male <55 u/L

enumerate the reference values of GGT according to IFCC

female 4-22 u/L, male 3-22 u/L

for 1-12 years old, what is the reference values of GGT according to IFCC?

female 4-24 u/L, male 2-42 u/L

for 13-18 years old, what is the reference values of GGT according to IFCC?

80-85%

the main part of phosphorus of the human body is that, how much of it is found in the bones?

inorganic phosphate

the remaining phosphorus are mainly?

calcium

what is another element that is in relation with phosphate in the human serum?

a decrease of the other component

an increase of one of these components usually leads to?

increase

if calcium decrease, what happens to phosphate?

vitamin D intoxication, hypoparathyroidism, renal failure

what are the conditions causes the elevation of serum phosphate to occur?

vitamin D deficiency, hyperparathyroidism

what are the conditions causes the decrease of serum phosphate to occur?

measuring phosphaturia

what can be proposed as a supplementary examination in serum phosphate?

during hyperparathyroidism, damage to the renal tubule in hypoparathyroidism

phosphaturia can be increase in what conditions?

Fanconi syndrome

what do you call the condition in damage to the renal tubule in hypoparathyroidism

phosphomolybdate UV endpoint

what is the method of phosphorus?

ammonium molybdate + sulfuric acid > phosphorus > phosphomolybdate complex

what is the principle of phosphorus?

phosphorus liqui-UV reagent

what is phosphorus reagent 1 called?

ammonium molybdate 0.8 mml/L with reaction accelerator in acidic solution

what compromises phosphorus liqui-UV reagent (R1)?

phosphorus standard

what is phosphorus reagent 2 called?

aqueous solution of monopotassium phosphate with added preservative

what compromises phosphorus standard (R2)?

2-25C, protect it from light, do not freeze

at what temperature are phosphorus reagent stored and what are their other conditions?

deteriorated

phosphorus reagents should be clear, if cloudy, the reagent has?

ready to use now

phosphorus reagent and standards are?

serum, lithium heparinized plasma (non-hemolyzed), urine

what are the acceptable specimens of phosphorus?

HCl to acidify (pH 5) after collection, dilute at 1/10 with saline solution (NaCl 9g/L)

what is the preparation of urine before it is analyze for phosphorus?

citrate, oxalate and EDTA

what anticoagulants are contraindicated from phosphorus analysis?

because it interferes with the formation of phosphomolybdate complex

why are anticoagulants contraindicated from phosphorus analysis?

4 days at room temp. 5 days at 2-8C, several months at -20C

phosphorus plasma and serum are stable up to?

6 months

acidified urine samples are stable up to?

340 nm

what is the wavelength serum phosphate read at?

mg/dL x 0.25 = mmol/L

what is the conversion factor of phosphorus?

adult 2.5-4.8 mg/dL, children (1st year) 4.0-7.0 mg/dL

enumerate the reference values of serum/plasma phosphate?

adult 0.34 - 1.0 gm/24 hours, children 0.54-0.84 gm/24 hours

enumerate the reference values of urine phosphate?

1 weak between -20 and 25C

In GGT specimen, how stable is it up to?

once, contaminated

In GGT specimen should be freeze how many time?, and discard if?