6, Rue Ibn Ennafis - Z.I. Lac 3 Tunisie

Tél. : 71 182 500 - Fax : 71 182 250

www.biomaghreb.com

CLINICAL SIGNIFICANCE

In humans, 70% of the body’s iron is bound to haemoglobin, the rest is bound to transport proteins

such as ferritin (or transferrin), or stored in certain tissues such as the liver and bone marrow. Low

serum iron levels may be seen in iron deficiency anemia (martial deficiency) or in patients with

inflammatory anemia.

Conversely, iron overload may occur during hemochromatosis or acute liver dysfunction.

PRINCIPLE

At pH 4.8 ferric iron (Fe

+++

) is instantly released from transferrin. Ascorbic acid reduces it to ferrous

iron (Fe

). Ferrozine forms with ferrous iron a soluble colored complex, measurable from 560 to

580 nm.

The presence of thio urea eliminates the interference of cuprous ions.

REAGENT COMPOSITION

Reagent 1

Guanidine, HCI

Acetate Buffer

4,5 mmol/l

pH 5

Reagent 2 Ascorbic acid 40 mmol/l

Reagent 3 Ferrozine 1 mg/l

Reagent 4 Standard 17,9 µmol/l

SAFETY CAUTIONS

Biomaghreb reagents are intended for use by qualified personnel for in vitro use (do not pipette by

mouth).

• Refer to the current MSDS available on request or on www.biomaghreb.com.

• Check the integrity of the reagents before use.

• Disposal of waste: comply with applicable legislation.

For safety reasons, treat any specimen or reagent of biological origin as potentially infectious. Ob-

serve the applicable legislation.

REAGENT PREPARATION

Dissolve the contents of a spatula of ascorbic acid (approximately 250 mg) in 50 ml of Reagent 1

(Reagent A).Add 40 µI ferrozine to 1 ml of Reagent A (Reagent B).Reagent B is prepared extempo-

raneously.

SAMPLE COLLECTION AND HANDLING

Serum,plasma heparinazed and non hemolysed

PRESERVATION AND STABILITY

• Before opening: Until the expiry date indicated on the label of the box at + 4°C.

• After opening : Reagents A and B are stable : 3 days at 20 - 25°C;

2 weeks at 2 - 8°C.

ADDITIONAL EQUIPMENT

• Basic equipment of the medical analysis laboratory ;

• Spectrophotometer or Clinical Biochemistry Analyzer.

LIMITS

The use of glass materials requires soaking for several hours in 2N hydrochloric acid and then rinsing

carefully with distilled water. It is therefore preferable to use disposable plastic material. High doses

of anticoagulants (Heparin) can cause disturbances in the reaction mixture.

QUALITY CONTROL

External quality control program.

It is recommended to control in the following cases:

• At least one test per series.

• Change of reagent bottle.

• After maintenance work on the analyzer.

If a control value is outside the confidence limits, repeat the procedure using the same control.

Use normal and pathological control sera.

CALIBRATION

• Standard (Reagent 3)

The calibration frequency depends on the performance of the analyzer and the storage conditions

of the reagent.

Recalibration is recommended in the following cases:

1. changing the reagent lot ;

2. after maintenance work on the analyzer; and

3. the control values are outside the confidence limits.

LINEARITY

The reaction is linear up to 1000 µg/dl (179.7 µmol/l).Above this limit, dilute the sample with 9 g/L

NaCl solution and repeat the determination taking the dilution into account in the calculation of the

result. The linearity limit depends on the sample/reagent volume ratio.

PROCEDURE

Wavelength: 578 nm (530-590).

Temperature: 20 -25°C.

Tank: 1 cm thick.

Adjust the spectrophotometer zero by :

• Reagent A for Sample Whites ;

• Reagent blank Reagent for standard and samples

Blank

Reagent

Standard

white

Sample

Distilled water

Sample

Reagent A

Reagent B

200µl

1ml

200µl

1ml

200µl

1ml

200µl

1ml

Mix and wait 10 minutes

Measuring absorbances at 578 nm

Staining stability: 30 minutes.

CALCULATION

Serum iron =

DO. Sample - DO. White sample

DO Standard

n= 1 mg/l;

n = 17, 9 µmol /l.

REFERENCE VALUES

Women

69 - 158 µg/dl

12.5 - 28.3 µmol/l

Men

59 - 145 µg/dl

10.7 - 26 µmol/l

REFERENCES

Persijn et al Clin. Chem. Acta 35,91 (1971);

Stoockey L. Anal. Chem. 42,779 (1970);

Williams et al. Clin. Chem. 23,237(1977).

IRON

Colorimetric Method

Quantitative determination of iron in human plasma

FT En 03 Date de MAJ : Janvier 2020 Version B

IN VITRO DIAGNOSTIC USE