IRON FERROZINE
REF 1135005
2 x 50 mL IRON
CONTENTS FERROZINE
R1. Reagent 2 x 40 mL Colorimetric method
R2. Reagent 1 x 20 mL ENDPOINT
CAL. Standard 1 x 3 mL
For in vitro diagnostic use only
PRINCIPLE INTERFERENCES
The Fe+3 bound to serum ferritine once dissociated in a week-acid Lipemia (intralipid 1.25 g/L) may affect the results.
medium by Teepol and guanidium chloride, is reduced by Bilirubin (< 40 mg/dL) does not interfere.
hydroxylamine to Fe+2, forming the ferrous ion a colored complex
Hemoglobin may affect the results.
with FerroZine® proportional to the concentration of iron present in
the sample.1,2 Other drugs and substances may interfere3.
pH 5
Transferrin -Fe+3 Apotransferrin + Fe+3
Teepol MATERIALS REQUIRED
Fe+3 + Hydroxylamine Fe+2
Photometer or colorimeter capable of measuring at 560 20 nm.
Fe+2 + FerroZine Fe (FerroZine)3+2 complex
Pipettes with disposable plastic tips to measure reagents and
samples.
Trade mark of Hach Chemical Co. Ames, Iowa.
Disposable plastic tubes for the tests.
REAGENT COMPOSITION
PROCEDURE
R1 Buffer/Reductant. Guanidine chloride 1.0 mol/L, hydroxylamine
0.6 mol/L, acetate buffer 400 mmol/L pH 4.0, Teepol.
1. Bring reagents and samples to room temperature.
R2 Chromogen. FerroZine 8 mmol/L, sodium acetate 400 2. Pipette into labelled test tubes:
mmol/L.
CAL Iron standard. Ferric ion 100 g/dL (17.9 mol/L). Reagent Sample CAL.
TUBES Sample
Concentration value is traceable to Standard Reference blank blank Standard
Material NIST 937.
Distilled water 200 L
Sample 200 L 200 L
STORAGE AND STABILITY [Link] 200 L
R1 1.0 mL
Store at 2-8ºC. Working reagent 1.0 mL 1.0 mL 1.0 mL
All the kit compounds are stable until the expiry date stated on the
label. Do not use reagents over the expiration date.
Store the vials tightly closed, protected from light and prevented 3. Mix and let the tubes stand 5 minutes at room temperature.
contaminations during the use. 1
Discard If appear signs of deterioration: 4. Read the absorbance (A) of the sample blank at 560 nm
- Presence of particles and turbidity. against distilled water.
- Blank absorbance (A) at 560 nm 0.050 in 1cm cuvette. 5. Read the absorbance (A) of the samples and the standard at
560 nm against the reagent blank.
REAGENT PREPARATION
CALCULATIONS
Working reagent. Mix 4 volumes of R1 + 1 volume of R2. Stable 6
months at 2-8ºC.
A Sample - A Sample blank
SAMPLES x C Standard = g/dL iron
A Standard
Serum or heparinized plasma. Centrifuge specimen as soon as
possible after collection. Samples with concentrations higher than 1000 g/dL should be
Hemolyzed samples are rejected. Ruptured red cells falsely elevate diluted 1:2 with saline and assayed again. Multiply the results by 2.
the serum results.
Iron in serum is stable for 3 weeks at 2-8ºC and for about 7 days at If results are to be expressed as SI units apply:
20-25ºC. Freeze for longer storage. g/dL x 0.179 = µmol/L
QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: info@[Link] ; website: [Link] NIF-VAT:B60485687
REFERENCE VALUES4 ANALYTICAL PERFORMANCE
Serum
- Detection Limit : 2.69 µg/dL
Men 60 - 175 g/dL (10.7 - 31.3 mol/L) - Linearity : Up to 1000 µg/dL
- Precision:
Women 50 - 170 g/dL (9.0 - 30.4 mol/L)
µg/dL Within-run Between-run
It is recommended that each laboratory establishes its own Mean 106 144.7 106 144.7
reference range
SD 2.33 2.70 2.17 2.03
QUALITY CONTROL CV% 2.19 1.87 2.04 2.09
N 10 10 10 10
The use of a standard to calculate results allows to obtain an
accuracy independent of the system or instrument used. - Sensitivity : 0.8 mAbs / µg/dL iron.
To ensure adequate quality control (QC), each run should include a
- Correlation: This assay (y) was compared with a similar commercial
set of controls (normal and abnormal) with assayed values handled
method (x). The results were:
as unknowns.
N = 62 r = 0.94 y = 0.97x + 0.1
REF 1980005 HUMAN MULTISERA NORMAL
The analytical performances have been generated using on
Borderline level of iron. Assayed.
automatic instrument. Results may vary depending on the
instrument.
REF 1985005 HUMAN MULTISERA ABNORMAL
Elevated level of iron. Assayed.
REFERENCES
If the values are found outside of the defined range, check the
instrument, reagents and procedure. 1. Carter, P. Anal. Biochem. 40 : 450 (1971).
Each laboratory should establish its own Quality Control scheme 2. Artiss, J.D., Vinogrador, S., and Zak, B. Clin. Biochem. 14 :
and corrective actions if controls do not meet the acceptable 311 (1981).
tolerances. 3. Young DS. Effects of drugs on clinical laboratory tests, 5th ed.
AACC Press, 2000.
4. Tietz, N.W., Fundamentals of Clinical Chemistry, p.940. W.B.
CLINICAL SIGNIFICANCE
Saunders Co., Philadelphia , 1987.
Following intestinal absorption of iron or erythrocyte destruction,
iron ions are released into the plasma where they bind to either
apotransferrin or apoferritin proteins to form transferrin and ferritin,
respectively. The former helps transport iron to bone marrow for
erythropoiesis; the latter stores iron in tissues, until is needed.
An increase in the iron level in plasma due to rapid destruction of
erythrocytes or excesive uptake of iron may also lead to iron
overload. The latter causes iron deposition disorders in tissue
known as hemosiderosis or hemochromatosis.
Conversely, a decrease in the iron level in plasma due to
malnutrition or malapsorption may lead to excesive depletion in iron
storage, resulting in anemia such as iron-deficiency anemia.
NOTES
Contamination of glassware with iron will affect the test. Use
acid-washed glassware or plastic tubes.
This method may be used with different instruments. Any
application to an instrument should be validated to demonstrate
that results meet the performance characteristics of the
method. It is recommended to validate periodically the
instrument. Contact to the distributor for any question on the
application method.
Clinical diagnosis should not be made on findings of a single
test result, but should integrate both clinical and laboratory
data.
B1135-3/1504
[Link]
QUALITY SYSTEM CERTIFIED LINEAR CHEMICALS, S.L.U. Joaquim Costa 18 2ª planta. 08390 Montgat (Barcelona) SPAIN
ISO 9001 ISO 13485 Telf. (+34) 934 694 990; E-mail: info@[Link] ; website: [Link] NIF-VAT:B60485687
