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DISCRETPAK™ REAGENTS - Total Iron

Intended Use

For IN VITRO diagnostic use in the quantitative determination of Total Iron in serum using manual or automated applications.

Method Principle

In acid buffer, Transferrin-bound iron dissociates into free Ferrous (Fe++) and Ferric (Fe+++) iron. Hydroxylamine hydrochloride present in the same acid reagent reduces the Ferric iron to the Ferrous state. The free Ferrous ions react with Ferene to form a complex with maximum absorbance at 600 nm. The absorbance is directly proportional to the concentration of total iron in the serum sample. The reaction scheme below illustrates the reaction that occurs in this method.

Hydroxylamine HCl
  Transferrin 2(Fe3+) ──────────────> Transferrin + Fe2+
pH 4.5

 Fe2+ + 3 Ferene ──────> Ferrous-Ferene Complex


Method Performance Characteristics

Sensitivity: The sensitivity of this method is 0.00010-0.00012 absorbance units per µg/dL.
Linear Range: In this method there is no significant nonlinearity over the range of 0-1200 µg/dL.
Precision: Within-run and day-to-day precision is summarized below.

Total Iron

Within-Run Precision

Total Precision

MEAN

SD

CV

SD

CV

µg/dL

µg/dL

%

µg/dL

%

25

2.60

*

3.40

*

477

7.40

1.50

8.20

1.70

948

5.10

0.50

14.80

1.50

*CV’s not meaningful when the average approaches zero.

Correlation

A comparison of this method using an automated analyzer was carried out against a reference method based on the ferene reaction. The performance of Catachem Total Iron method was found essentially equivalent to the reference method used. Linear regression analysis produced the following information:

Correlation Data

Parameter

Data Observed

N

93

Range (µg/dL)

7-245

Regression

Y = 0.989x + 2.0

Correlation

r = 0.961

Sy,x

9.9