For the quantitative determination of iron and total iron-binding capacity in serum.
PRINCIPLE
Total Iron-Binding Capacity (TIBC): A known amount of ferrous ions are added to serum at an alkaline pH. The ferrous ions bind with transferrin at unsaturated ironbinding sites. 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 (UIBC). The TIBC is equal to the serum iron concentration plus the UIBC.
REAGENTS
1. UIBC/TIBC BUFFER REAGENT: Tris 500mM, pH 8.1 with surfactant, Sodium Azide 0.05% (w/v) as preservative.
2. UIBC/TIBC COLOR REAGENT(Cromogen) : Ferrozine in hydroxylamine hydrochloride.
3. UIBC/TIBC STANDARD (500 g/dL): Ferrous chloride in hydroxylamine hydrochloride.
Precautions
1. All reagents are toxic. Do not pipette by mouth. Avoid all contact.
2. UIBC buffer contains sodium azide and may react with lead and copper plumbing to form highly explosive metal azides.
3. On disposal, flush with a large volume of water to prevent azide accumulation.
4. This reagent is for in vitro diagnostic use only.
REAGENT DETERIORATION
All reagents should be clear. Turbidity may indicate contamination and the reagent should not be used.
SPECIMEN COLLECTION AND STORAGE
1. Fresh, unhemolyzed serum is the specimen of choice.
2. Serum should be separated as soon as clot has formed.
3. Heparinized plasma may be used but other anticoagulants should not be used to avoid possible iron contamination.
INTERFERENCES
1. Certain drugs and other substances are known to influence circulating iron levels.