C liquicolor Calculation

Photometric Colorimetric Test for Chloride Asample

c = 100 x  [mmol/l]
TPTZ Method A[STD]
Package Size or
[REF] 10115 200 ml Complete test kit Asample
c = 355 x  [mg/dl]
[IVD]
A[STD]
Method1,2 The result may be converted according to the following equations:
Chloride ions react with a mercury(II)-2,4,6-tri-(2-pyridyl)-s-triazine (TPTZ) c [mmol/l] = c [mval/l]
complex to form mercury(II)-chloride. The liberated TPTZ reacts with
iron(II) ions yielding a blue coloured complex. The resulting absorbance c [mmol/l] x 3.55 = c [mg/dl]
change at 590 nm is directly proportional to the amount of chloride ions c [mg/dl] x 0.282 = c [mmol/l]
in the sample.
24 h urine: c [mmol/l] x urine volume [l] / 24 [h] = c [mmol/24 h]
Contents c [mmol/24 h] x 0.0355 = c [g/24 h]
[RGT] 2 x 100 ml Colour reagent
2,4,6-Tri-(2-pyridyl)-s-triazine (TPTZ) Performance Characteristics
(partially as mercury(II) complex) 0.986 mmol/l Linearity
Iron(II) sulphate 0.53 mmol/l The test is linear up to a chloride concentration of 500 mmol/l or 1775
[STD] 3 ml Standard mg/dl. Samples with higher chloride concentration have to be further
Chloride (Cl-) 100 mmol/l diluted 1 + 1 with distilled water.
or 355 mg/dl Repeat the assay and multiply the results by 2.
Reagent Preparation Typical performance data can be found in the Verification Report,
[RGT]and are ready to use.
[STD] accessible via:
www.human.de/data/gb/vr/ey-cl.pdf or
Reagent Stability
[RGT]and [STD] are stable even after opening up to the stated expiry date www.human-de.com/data/gb/vr/ey-cl.pdf
when stored at 2...25°C in the dark. Contamination must be avoided. Normal Values
Specimen Serum: 95 - 108 mmol/l 335 - 383 mg/dl
Serum, CSF, urine CSF: 119 - 130 mmol/l 442 - 460 mg/dl
Stability in serum at 2...25°C: 7 days Urine: 110 - 225 mmol/l 3.9 - 8 g/24 h
Stability in urine at 2...8°C: 7 days Quality Control
Notes All control sera with chloride values determined by this method can be
Blood should be immediately processed (e.g. centrifuged within 1 hour), employed.
otherwise an electrolyte exchange between serum and erythrocytes may We recommend to use our quality control sera HumaTrol based on
lead to a false low chloride determination. animal serum or our SERODOS based on human serum.
Precipitates in urine may be redissolved by warming up. Automation
Assay Proposals to apply the reagents on analyzers are available on request.
Wavelength: 590 nm (560 - 600nm), Hg 578 nm Each laboratory has to validate the application in its own responsibility.
Optical path: 1 cm Notes
Temperature: 20...37°C 1. The test detects chloride very sensitively. Usage of contaminated
Measurement: Against reagent blank. Only one blank per series is glassware and skin contact therefore can cause elevated chloride re-
required. sults. Use of disposable plastic ware and gloves is highly
recommended.
Pipetting Scheme 2. Hemoglobin, bilirubin and ascorbic acid in physiological concentrations
Macro method: Predilute and the sample 1 + 40 with distilled water,
[STD] do not interfere with the test. Lipemic samples interfere with the test
e.g. 50 µl[STD] /sample + 2000 µl dist. water. (elevated results) and should therefore be avoided.
Semi-micro method: Predilute and the sample 1 + 50 with distilled
[STD] 3. [RGT] is sensitive to light, especially UV light. should be kept
[RGT]

water, e.g. 20 µl [STD] /sample + 1000 µl dist. water. light-protected.

Use the prediluted /sample for the following pipetting scheme.
[STD]
References
Pipette into Macro Semi-micro 1. Prellwitz W.; Klinisch-chemische Diagnostik, Thieme Stuttgart, 2. ed.,
1976
cuvettes [STD] Sample [STD] Sample 2. Fried R.et al.,J. Clin. Chem. Clin. Biochem. 10, 280 (1972)
50 µl --- 20 µl ---
|
[STD]

Sample --- 50 µl --- 20 µl EY-CL INF 1011501 GB 06-2008-9

[RGT] 2000 µl 2000 µl 1000 µl 1000 µl
Mix, incubate for 5 minutes in the dark and measure the absorbance of
sample (Asample) and (A[STD])within 60 minutes against the reagent
[STD]

blank. Do not expose to light!

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