!+?,@?+4168
!"#$%&'()!"#$"%&!&'
Photometric Colorimetric Test for Chloride
TPTZ Method
or
*+,-+./)012/
3
10115
!"#$%
!&'(%
Asample
c = 100 x !!!! [mmol/l]
A!+*(%
200 ml
Complete test kit
Asample
c = 355 x !!!! [mg/dl]
A!+*(%
3
The result may be converted according to the following equations:
3/4567)1,2
Chloride ions react with a mercury(II)-2,4,6-tri-(2-pyridyl)-s-triazine
(TPTZ) complex to form mercury(II)-chloride. The liberated TPTZ
reacts with iron(II) ions yielding a blue coloured complex. The resulting
absorbance change at 590 nm is directly proportional to the amount of
chloride ions in the sample.
!684/849
!")*% :);)<==)>?)!6?6@A)A/+./84
2,4,6-Tri-(2-pyridyl)-s-triazine (TPTZ)
(partially as mercury(II) complex)
Iron(II) sulfate
0.986 mmol/l
0.53 mmol/l
!+*(% B)>?)04+87+A7
Chloride (Cl-)
or
100 mmol/l
355 mg/dl
%/+./84)*A/C+A+4168
!")*% and !+*(% are ready to use.
%/+./84)04+D1?1414E
!")*% and !+*(% are stable even after opening up to the stated expiry
date when stored at 2...25°C in the dark. Contamination must be
avoided.
0C/,1>/8
Serum, CSF, urine
Stability in serum at 2...25°C:
Stability in urine at 2...8°C:
7 days
7 days
F64/9
Blood should be immediately processed (e.g. centrifuged within 1
hour), otherwise an electrolyte exchange between serum and erythrocytes may lead to a false low chloride determination.
Precipitates in urine may be redissolved by warming up.
G99+E
Wavelength:
Optical path:
Temperature:
Measurement:
590 nm (560 - 600nm), Hg 578 nm
1 cm
20...37°C
Against reagent blank. Only one blank per series
is required.
*1C/4418.)0,5/>/
3+,A6)>/4567H Predilute !+*(% and the sample 1 + 40 with distilled
water, e.g. 50 µl !+*(%/sample + 2000 µl dist. water.
0/>1I>1,A6) >/4567H Predilute !+*(% and the sample 1 + 50 with
distilled water, e.g. 20 µl !+*(%/sample + 1000 µl dist. water.
c [mmol/l]
c [mmol/l] x 3.55
c [mg/dl] x 0.282
=
=
=
c [mval/l]
c [mg/dl]
c [mmol/l]
24 h urine: c [mmol/l] x urine volume [l] / 24 [h]
c [mmol/24 h] x 0.0355
=
=
c [mmol/24 h]
c [g/24 h]
*/AJ6A>+8,/)!5+A+,4/A1941,9
Linearity
The test is linear up to a chloride concentration of 500 mmol/l or 1775
mg/dl. Samples with higher chloride concentration have to be further
diluted 1 + 1 with distilled water.
Repeat the assay and multiply the results by 2.
Typical performance data can be found in the Verification Report,
accessible via:
www.human.de/data/gb/vr/ey-cl.pdf or
www.human-de.com/data/gb/vr/ey-cl.pdf
F6A>+?)K+?@/9
Serum:
95 - 108 mmol/l
CSF:
119 - 130 mmol/l
Urine:
110 - 225 mmol/l
335 - 383 mg/dl
442 - 460 mg/dl
3.9 - 8 g/24 h
L@+?14E)!684A6?
All control sera with chloride values determined by this method can be
employed.
We recommend to use our animal serum based "M3GN%$# quality
control sera or our human serum based 0(%$'$0.
G@46>+4168
Proposals to apply the reagents on analysers are available on request.
Each laboratory has to validate the application in its own responsibility.
F64/9
1. The test detects chloride very sensitively. Usage of contaminated
glassware and skin contact therefore can cause elevated chloride
results. Use of disposable plastic ware and gloves is highly
recommended.
2. !")*% is sensitive to light, especially UV light. !")*% should be kept
light-protected.
%/J/A/8,/9
1. Prellwitz, W.; Klinisch-chemische Diagnostik, Thieme Stuttgart,
2. ed., 1976
2. Fried, R. !"#$%., J. Clin. Chem. Clin. Biochem. <=, 280 (1972)
3. ISO 15223 Medical devices – Symbols to be used with medical
device labels, labelling and information to be supplied.
Use the prediluted !+*(%/sample for the following pipetting scheme.
Pipette into
cuvettes
!+*(%
Sample
!")*%
Macro
Semi-micro
!+*(%
Sample
!+*(%
Sample
50 µl
--2000 µl
--50 µl
2000 µl
20 µl
--1000 µl
--20 µl
1000 µl
Mix, incubate for 5 minutes in the dark and measure the absorbance
of sample (Asample) and !+*(% (A!+*(%) within 60 minutes against the
reagent blank. Do not expose to light!
EY-CL
INF 1011501 GB
12-2003-6
!
Human Gesellschaft für Biochemica und Diagnostica mbH
Max-Planck-Ring 21 - D-65205 Wiesbaden - Germany
Telefon: +49 6122 9988 0 - Telefax: +49 6122 9988 100 - eMail: human@human.de