IRON-FS (5+1)
(Colorimetric Test
with Ferene-S)
METHOD
Ferene. End Point.
REFERENCE VALUES
µg/dl
PRINCIPLE
At pH 4.5 ferric iron is released from the transferrin
complex. Ascorbic acid reduces the ferric iron to
ferrous state which reacts with Ferene to produce a
colored complex.
REAGENTS
Reagentcomposition
R1:
Bufferreagent
R2:
R3:
Acetate buffer, pH 4.50
800
Thiourea
90
Ascorbic acid (spoon supplied) 45
Colorreagent
Ferene
0,6
Ascorbic acid
(spoon supplied)
Standardsolution
Iron ions
mmol/L
mmol/L
mmol/L
mmol/L
45
mmol/L
100
17.9
µg/dL
µmol/l
Precautions
- For in vitro diagnostic use only.
- Avoid contamination by using clean laboratory
material (pipette, plastic vial for analyzers,...).
Stability
When stored at 2-8°C and protected from light, the
reagents are stable up to l the expiry date stated on
the labels.
Preparation of Working Reagent / Stability
Working reagent (WKR1) :
Dissolve 1 spoonful (~ 100 mg) of ascorbic acid in 20
to 25 mL of buffer R1.
Stability :
3 days at 20-25°C
2 weeks at 2-8°C
R2 is ready for use.
SAMPLES
Serum. Heparin plasma.
(Discard hemolyzed samples)
Children
2 Weeks
6 Months
12 Months
12 Months
2 –12 Years
Women
25 Years
40 Years
60 Years
Pregnant Women
12. week
At birth
6 weeks post partum
Men
25 Years
40 Years
60 Years
µmol/l
63-201
28-135
35-155
35-155
22-135
11-36
5-24
6-28
6-28
4-24
37-165
23-134
39-149
6,6-29,5
4,1-24,0
7,0-26,7
42-177
25-137
16-150
7,6-31,6
4,5-24,5
2,9-26,9
40-155
35-168
40-120
7,2-27,7
6,3-30,1
7,2-21,5
Note: It is recommended for each laboratory to establish its
own reference values. The given data are only an indication.
ASSAY PROCEDURE
Reagent can be used manually (see method
below) and on most analyzers. Applications are
available on request
Startreagentprocedure:
Wavelength :
Temperature :
Cuvette
:
600 nm, 578 nm
25°C, 37°C
1 cm light path
RB
Calibrator SAMPLE
WKR 1
1 mL
1 mL
1 mL
Distilled water
200 µL
Calibrator
200 µL
Sample
200 µL
Mix and read Absorbance A1 of Reagent Blank, Sample
and Calibrator, then add:
R2
200 µL
200 µL
200 µL
Mix and read Absorbance A2
A= [(A2 – 0.82 A1) Sample/Cal] – [(A2 - 0.82 A1) RB]
CALCULATION
As / Ac x C
(C = Calibrator concentration)
Page 1/2
Greiner Diagnostic GmbH - Unter Gereuth 10 – D-79353 Bahlingen – Germany
www.greiner-diagnostic.com
CALIBRATION & QUALITY CONTROL
For the calibration of automated analyzers
Greiner Multicalibrator is recommended, for
quality control use Greiner normal and
abnormal control, Unitrol I and Unitrol II.
BIBLIOGRAPHY
1.
2.
PERFORMANCE DATA (37°C)
- Analytical range
The reagent is linear up to 1100 µg/dL
3.
4.
- Detection limit
The detection limit is 1.4 µg/dl
5.
- Precision
Within-run reproducibility
N = 11
Mean
SD
CV
Unit
Control 1
112,5
1,44
1,28%
µg/dl
Control 2
173,9
2,43
1,40%
µg/dl
Patient
55,5
2,60
4,68%
µg/dl
Wick M. Iron metabolism and its disorders. In:
Thomas L, editor. Clinical laboratory diagnostics. 1st
ed. Frankfurt: TH-Books Verlagsgesellschaft; 1998.
p. 268-73.
Fairbanks VF, Klee GG. Biochemical aspects of
hematology. In: Burtis CA, Ashwood ER, editors. Tietz
Textbook of Clinical Chemistry. 3rd ed. Philadelphia:
W.B Saunders Company; 1999. p. 1642-1710.
Higgins T. Novel chromogen for serum iron
determinations. Clin Chem 1981;27:1619.
Artiss JD, Vinogradov S, Zak B. Spectrophotometric
study of several sensitive reagents for serum iron.
Clin Biochem 1981;14:311-15.
Thomas L. Clinical Laboratory Diagnostics. 1st ed.
Frankfurt: TH-Books Verlagsgesellschaft; 1998.
p. 273-5
SYMBOLS USED
Between-run reproducibility
N = 11
Mean
SD
CV
Unit
Control 1
110,7
1,47
1,33%
µg/dl
Control 2
177,9
1,97
1,11%
µg/dl
Patient
54,5
1,21
2,22%
µg/dl
- Correlation
A comparative study has been performed
between the Greiner method and another
commercial reagent on 23 human serum
samples. The parameters of linear regression
are as follows:
y = 1.065 x - 0.692 µg/dl
For in vitro diagnostic medical use
Batch Code
Use by
Temperature limitation
r = 0.994
-Interferences
Interferences are found according to literature
Page 2/2
Greiner Diagnostic GmbH - Unter Gereuth 10 – D-79353 Bahlingen – Germany
www.greiner-diagnostic.com