CHOLRIDE REAGENT
Mercurous (II) thiocyanate method
Quantitative determination of Chloride in serum/plasma Sample collection and preservation
Only for In Vitro Diagnostic use Serum or plasma(lithium heparin)
Separate from cellular contents immediately after blood colletion.
Clinical significance Stability: at least one year at -20º C in case of immediate freezing.
It is important clinically the determination of chloride due regulation 7 days at 4-8ºC freeze only once! Discard contaminated
of osmotic pressure of extra cellular fluid and to its significant role specimens!
in acid- base balance. Increases in chloride ion concentration may Urine: colletc 24-hour urine specimen in chloride free
be found in severe dehydration, excessive intake of chloride, [Link] a sample ½ in distilled [Link] results by
severe renal tubular damage and in patients with cystic 2 (dilution factor).
fibrosis. Decrease in chloride ion concentration may be found in Stability of the sample: 1 week at refrigerator (2-8) or frozen(-20ºC)
metabolic acidosis, loss from prolonged vomiting and chronic temperatures.
pyelonephritis. Clinical diagnosis should not be made on a
single test result; it should integrate clinical and other - Proceed carefully with this product because due to its nature
laboratory data. it can get contaminated easily.
- most of the detergents and water softening products used in
Method the laboratories contain chelating agents. A defective rising
Photometric test using mercorous (II) thiocyanate. will invalidate the procedure.
Principle ASSAY PROCEDURE
Chloride ions react with mercurous thiocyanate to form Operating instructions
mercury per chlorate and thiocyanate. Thiocyanate forms a red - Check reagent inventories at least daily to ensure that
complex with ferric ions in the presence of nitric acid. quantities are sufficient for the palnned work load.
- Bring all reagents,standar and sample to room temperature
18-28ºC,prior to analysis.
Reagent composition
Reagent I : Chloride Reagent Automated parameters
Chloride standard : 100 mEq/L Wavelength 505nm
Cuvette 1 cm 1 cm
Reagent preparation Reaction temperature R.T R.T (22-30ºC)
All reagent is provided in ready to use. Measurement against Reagent blank
Reaction Direction increasing
Reagent storage and stability Reaction type End point
When stored at recommended storage temperature stated on Sample volumen 10 µl
label,reagent is stable until the expiration date stated o the bottle Reagent volume 1000 µl
and kit box label. Incubation 5 min.
Blank Absorbance limit ˂ 0.30
Warning and precautions Units mEq/L
- For in vitro diagnostic use
- Do not use components beyond the expiration date.
- Do not mix materials from different kit lot numbers. Manual assay procedure
- Exercise the normal precautions required for handling all Pipette into test tubes
laboratory reagents.
- The reagent contains preservative. Do not swallow. Avoid
Blank Standard Test
contact with skin and mucous membranes.
Reagent 1 1000 µl 1000µl 1000µl
- For detailed information refer material safety data sheet.
Standard - 10 µl -
- Proceed carefully whith this product because due to its nature
it can get contaminated easily. Serum / plasma - - 10 µl
- Most of the detergent and water softetening products used in
the laboratories contain chelating agents. A defective rinsing - Mix & incubate for 5 min at RT(22-30ºC)
will invalidate the procedure. - Read an record absorbance of the reagent blank,standar,
Wasted Management control and each unknow sample immediately.
Please refer to local legal requerements. - CHLOM 50 are specially treated monovials with 1ml pre-
Materials required but not provided dispensed [Link] add 10µl sample / std,incubate at R.T
- NaCl solution 9 g/l for 5 min.&[Link] the same programme as above.
- General laboratory equipment
Sample dilutions
Limitation of the procedure
- This method is linear upto a concentration of 130 mEq/L - for diagnostic purposes, the reults should always be assessed
- Dilute samples above this concentration 1:1 with DI water in conjunction with the patient`s medical history,clinical
- and repeat [Link] the result by 2. examination and other findings.
Interference
- Bilirubin: No interference found upto bilirubin 32mg/dl.
Calculation
- Hemoglobin: No interference found upto 500mg/dl
- Lipemia: No interference found upto 500mg/dl
Chloride (mEq/l) = AT/AS x Conc of Standard
- The characteristics have been obtained using an automatic
Chloride in Urine (mEq/l) = AT/AS x Conc of Standard x2
[Link] may vary if a different instrument or a
manual procedure is used.
Calibrators and controls
Bibliography
1. Tietz, N.W., white ,W.L Mosby CO [Link] [Link] D.S henry
For the calibration of automated photometric systems the
R,J chem (1964) 10, 533.
commercially available suitable multi-calibrator is recommended.
The assigned values of Chloride standard have been made
traceable to the NIST standard reference material SRM 956.
It is recommended to run a normal and a pathological control
serum which is commercially available to verify the performance of
the measured procedure. The value of controls should fall within te
established limit.
Each laboratory should establish corrective action n case of
deviations in control recovery.
Perfomance Characteristics
Within Run
Mean
Sample SD CV %
concentration
Norm 99.64 3.38 3.39%
path 128.56 4.21 3.27%
Run to Run
Mean
Sample SD CV %
concentration
Norm 99.62 2.24 2.35%
path 128.83 3.14 2.44%
Linearity
The method is linear to a concentration of 7 mEq/[Link] samples
above this concentration 1:1 with DI water and repeat
[Link] the result by 2.
Limit of detection: the limit of detection for Chloride is 30mEq/L
Method Comparison
A comparison of accucare Chloride with a commercially available
assay (x) using 20samples gave following results: R2 =0.9590
Reference interval
Serum 95-115 mEql/L
Urine 110-250mEq/L / 24hr
CSF 95-110 mEq/L
Sweat Upto 60 mEq/L
The reference values are to be considered as indicative [Link]
laboratory should establish its own normal range.