CHLORIDE

REF 1116005
2 x 50 mL CHLORIDE
THIOCYANATE
CONTENTS
Colorimetric method
R1. Reagent 2 x 50 mL
CAL. Standard 1 x 3 mL ENDPOINT
For in vitro diagnostic use only

PRINCIPLE INTERFERENCES

Chloride ions in the sample quantitatively displaces thiocyanate  Lipemia (intralipid 1 g/L) may affect the results.
from mercuric thiocyanate. Liberated thiocyanate ion reacts with  Bilirubin (40 mg/dL) does not interfere.
ferric ion forming a red ferric-thiocyanate complex proportional to  Hemoglobin (12 g/L) does not interfere.
the concentration of chloride present in the sample.1,2  Other drugs and substances may interfere3.

2 Cl- + Hg (SCN)2 HgCl2 + 2 (SCN)- MATERIALS REQUIRED

3 (SCN)- + Fe3+ Fe (SCN)3  Photometer or colorimeter capable of measuring absorbance at

470  10 nm.
 Constant temperature incubator set at 37ºC (optional).
REAGENT COMPOSITION
 Pipettes with disposable plastic tips to measure reagents and
samples.
R1 Thiocyanate reagent. Mercuric thiocyanate 2 mmol/L,  Disposable plastic tubes for the tests.
mercuric nitrate 0.1 mmol/L, iron nitrate 30 mmol/L,
HNO3 45 mmol/L. (see Notes). Xn
CAL Chloride / Phosphorus standard. Chloride 100 PROCEDURE
mEq/L / Phosphorus 5 mg/dL.
Organic matrix based primary standard. Concentration 1. Bring reagents and samples to room temperature.
value is traceable to Standard Reference Material 909b. 2. Pipette into labelled test tubes:

STORAGE AND STABILITY CAL.

TUBES Blank Sample
Standard
Store at 2-30ºC. R1.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. Sample  10 L 
Store the vials tightly closed, protected from light and prevented CAL.Standard   10 L
contaminations during the use.
Discard If appear signs of deterioration:
- Presence of particles and turbidity. 3. Mix gently by inversion one or two times. Do not shake or stir
- Blank absorbance (A) at 470 nm > 0.050 in 1cm cuvette. vigorously.
4. Incubate the mixture for 5 or 10 minutes at a selected constant
temperature between 25-37ºC (see Notes).
REAGENT PREPARATION 5. Read the absorbance (A) of the samples and the standard at
470  10 nm against the reagent blank.
The Reagent and Standard are ready-to-use.
The color is stable for about 2 hours, at room temperature,
protected from light.
SAMPLES
CALCULATIONS
Serum, heparinized plasma and CSF.
Serum, plasma
The sera are stable in capped tubes for at least 4 hours at room
temperature, 2 days refrigerated (4-8ºC) and several months A Sample
frozen (-20ºC).
x C Standard = mEq/L (mmol/L) chloride
Cerebrospinal fluid should be collected in three sterile tubes, with
samples from the first or second tube used for chloride A Standard
determinations.

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: [email protected] ; website: www.linear.es NIF-VAT:B60485687
Samples with concentrations higher than 125 mEq/L (125 mmol/L) - Sensitivity : 0.005A / mmol/L chloride.
should be diluted 1:2 with distilled water and assayed again. Multiply
the results by 2. - Correlation: This assay (y) was compared with a similar
Concentrations less than 75 mmol/L, should be brought within linear commercial method (x). The results were:
range by increasing the amount of serum used. N = 20 r = 0.993 y = 1.05x - 3.1
The analytical performances have been generated using on
REFERENCE VALUES2 automatic instrument. Results may vary depending on the
instrument.

Serum, plasma 98 - 111 mEq/L (98 - 111 mmol/L)

NOTES
CSF 120 - 130 mEq/L (120 - 130 mmol/L)
 The reagent is harmful if swallowed. Do not pipette by mouth.
It is recommended that each laboratory establishes its own In case of accident or if you feel unwell, seek medical advice
reference range. immediately.
 A 5-min incubation period is satisfactory when the incubation
and assay are performed at 37ºC. A 10-min incubation period
QUALITY CONTROL is recommended when the incubation and assay are performed
at room temperature.
The use of a standard to calculate results allows to obtain an  For optimum results, the glassware used for the chloride
accuracy independent of the system or instrument used. procedure should be scrupulously clean. It may be found
To ensure adequate quality control (QC), each run should include a convenient to acid wash the material (H2SO4-K2Cr2O7) and then
set of controls (normal and abnormal) with assayed values handled thoroughly rinse it with distilled water.
as unknowns.  This method may be used with different instruments. Any
application to an instrument should be validated to demonstrate
REF 1980005 HUMAN MULTISERA NORMAL
that results meets the performance characteristics of the
Borderline level of chloride. Assayed. method. It is recommended to validate periodically the
REF 1985005 HUMAN MULTISERA ABNORMAL instrument. Contact to the distributor for any question on the
Elevated level of chloride. Assayed. application method.
 Clinical diagnosis should not be made on findings of a single
If the values are found outside of the defined range, check the test result, but should integrate both clinical and laboratory
instrument, reagents and procedure. data.
Each laboratory should establish its own Quality Control scheme
and corrective actions if controls do not meet the acceptable
tolerances. REFERENCES

CLINICAL SIGNIFICANCE 1. Schoenfeld R.G., and Lewellen Clin. Chem. 10 : 533 (1964).
2. Tietz. N.W. Clinical Guide to Laboratory Tests, 3rd Edition. W.B.
Sodium and chloride represent the majority of the osmotically active Saunders Co. Philadelphia, PA. (1995).
constituents of plasma. As a result, chloride is significantly involved 3. Young DS. Effects of drugs on clinical laboratory tests, 5th ed.
in maintenance of water distribution, osmotic pressure, and anion- AACC Press, 2000.
cation balance in the extracellular fluid compartment.
Hypochloremia (decreased plasma Cl- concentration) is observed in
salt-losing nephritis as associated with chronic pyelonephritis. In
Addison´s disease, Cl- levels as well as Na+ levels may drop
significantly during the Addisonian crisis and in certain types of
metabolic acidosis (e.g., diabetic ketoacidosis and renal failure) and
aldosteronism. In metabolic alkalosis, plasma levels of Cl- tend to
fall while HCO3- levels increase.
Hyperchloremia (increased plasma Cl- concentration) occurs with
dehydration, renal tubular acidosis, diabetes insipidus, acute renal
failure, adrenocortical hyperfunction and metabolic acidosis.
Extremely high dietary in take of salt and overtreatment with saline
solutions are also causes of hyperchloremia.

ANALYTICAL PERFORMANCE

- Linearity : Between 70 and 125 mmol/L

- Precision:
mmol/L Intraserial Interserial
Media 75 100 120 75 101 121
DE 1.3 2.2 1.7 1.8 2.5 1.7
CV% 1.7 2.2 1.4 2.4 2.5 1.4
N 10 10 10 10 10 10
B1116-2/0901
R1.ing

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: [email protected] ; website: www.linear.es NIF-VAT:B60485687