Chloride ( Thiocynate Method )

 Clinical significance :-

                            Chloride is one of the most important in the blood.  It  helps  keep  the amount of  fluid inside and outside of your cells in balance . It also help maintain proper blood volume, blood pressure and pH of your body fluids. If chloride level are increased called hyperchloremia. If chloride level are decrease called hypochloremia .

Principle :-

                   Chloride ions combine with free mercuric ion and release thiocyanate from mercuric thiocyanate. The thiocyanate released combines with the ferric ions to form a red brown ferric thiocyanate complex. Intensity of the colour formed is directly proportional to the amount of chloride present in the sample.

 Material required :-

• Clean and dry glassware.                                     
• Laboratory glass Pipettes or  micro Pipettes and tips. 
• Colorimeter.

 Normal Value :- 

                             98  - 106 mEq/L   

Sample :-

           Serum.

Procedure :-

             Pipette into clean and dry test tubes labeled as Blank ( B ) , Standard ( S ), and Test ( T ) :

Addition           Sequence	B	S	T
Reagent	1ml	1ml	1ml
Standard	-	10 ul	-
Sample	-	-	10ul

 Mix well  and  incubate  at for 02  mins. Measure absorbance of the standard ( Abs . S ) and Test ( Abs . T ) against Reagent blank.

 

   Calculation  :-

                                              Abs . T

                    Choloride ( mEq/L ) =    ..............  * 100

                                              Abs . S   

Preparation and reagent and stability :-

                                        All Reagent are ready to use and are stable till the expiry date mentioned on the label , when stored at 2' to 8' C.

Quality Control :-

                  For accuracy it is necessary to run known controls with every assay.

Limitation & Precautions :-

 

1. Storage conditions as mentioned on the kit to be adhered.
2. Do not freeze or expose the reagent to high temperature and protect from direct as it may affect the performance of the kit.
3. Use clean glassware free from dust .
4. Before the assay bring all reagents to room temperature.

 General System Parameters:-

          Reaction type           : End point 

          Wave length             : 505 nm 

          Temperature             : 37'C

          Incubation              : 02 minutes 

          Reagent Volume          : 1.0 ml

          Sample Volume           : 10 ul 

          Standard concentration  : 100 mEq/l

          Zero setting            : Reagent Blank

          Light Path              : 1.0 cm