Calcium

  Clinical significance :-

                            Calcium, in the body ,is found mainly in the bones ( approximately 99%). In serum calcium exist equally in a free ionised form and in a bound form .Hence a decrease in albumin causes lower calcium levels and vice versa . The level of Calcium in serum depend on the parathyroid hormone. Increased Calcium level are found in bone tumors , hyperparathyroidism. Decreased level are found in hypothyroidism,renal failure,rickets,Vitamin D deficiency and pancreatitis.   If calcium  level are increased called hypercalcemia. If calcium level are decrease called hypocalcemia .

Principle :-

                   Calcium in an alkaline medium combines with o- Cresolphthalein Complexone to form a purple coloured complex. Intensity of the colour formed is directly proportional to the amount of calcium present in the sample.  

 Material required :-

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

 Normal Value :- 

                             9.0  - 10.6 mg/dl   

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	-	20 ul	-
Sample	-	-	20ul

 Mix well  and  incubate  at for 05  mins. Measure absorbance of the standard ( Abs . S ) and Test ( Abs . T ) against Reagent blank.

 

   Calculation  :-

                                              Abs . T.    Calcium ( mg/dl ) =    ..............  * 10

                                              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             : 578 nm 

          Temperature             : Room Temperature

          Incubation              : 05 minutes 

          Reagent Volume          : 1.0 ml

          Sample Volume           : 20 ul 

          Standard concentration  : 10 mg/dl

          Zero setting            : Reagent Blank

          Light Path              : 1.0 cm