Professional Documents
Culture Documents
Khouja
Clinical Correlations
• Hyperphosphataemia
o Hypervitaminosis D -Hypoparathyroidism -Renal failure
o Acromegaly -Diabetic acidosis -Intestinal
obstruction
o Non-pathological bone regrowth during healing of fracture
• Hypophosphataemia
o Prolonged vomiting -Prolonged diarrhea
o Vitamin D deficiency rickets -Osteomalacia
o Hyperparathyroidism
o Fanconi Syndrome (defect in the reabsorption of Pi & other metabolites by
the renal tubules)
o Malnutrition
o Renal tubular acidosis
o Treatment of diabetic acidosis
o Extended I.V infusion of dextrose 5%
o Ingestion of phosphate-binding antacids
o Therapy with
Acetazolamide
Insulin
1
Pi Dr H. Khouja
Adrenalin (epineherin)
o Non-pathological immediately after a meal due to its mobilization into
cells for energy requirements & intermediary metabolism
Determination of Phosphate
Specimen :
• Fasting blood serum. Separate serum as soon as clot forms. Analyze immediately or
freeze at -20oC if not assayed soon.
o If freezing is delayed, organic phosphate may dissociate to Pi thus falsely
increasing Pi
o Avoid haemolysis false increase in Pi
o Never collect after a meal decrease in Pi
o Check for any treatment or drug intake
• Urine 24hr sample
Methodology
Principle:
All the methods depend on the specific reaction of Pi with ammonium molybdate in acid medium.
The resulting complex mixture is then reacted with a reducing agent which usually produce a
coloured product which is read in the spectrophotometer. Colour intensity is directly proportional to
the concentration of Pi.
Absorbance α Pi Concentration
Notes:-
There are several reducing agents available such as; ascorbic acid; stannous
chloride, Fe2+, malachite green (wave length and colour are different for each reducing agent)
The pH of the reaction must be maintained at an acidic pH to prevent
dissociation of organic phosphate into Pi and falsely increase the result
Protein precipitation is quite important as it eliminates interference and
matrix effects
The addition of the reducing agent to the phosphomolybdate complex must
be rapid to prevent any dissociation of the phosphomolybdate and give false decreased results