Purpose of the Test
To determine the total calcium concentration in serum samples.
Principle
Calcium reacts with o-cresolphthalein complexone (CPC) under alkaline conditions to form a purple colored complex. The intensity of the color formed is proportional to the calcium concentration in the sample.
Equipments
1. Spectrophotometer
2. Cuvettes
3. Micropipettes
4. Test tubes
5. Timer
6. CPC reagent
Sample Type
Serum or plasma is acceptable for the Test
Procedure
1. in 3 clean dry tubes add reagents and sample as mentioned in the Table below
| Blank | Standard | Test | |
|---|---|---|---|
| Calcium Reagent | 1000 μl | 1000 μl | 1000 μl |
| STD | – | 10 μl | – |
| Sample (Serum) | – | – | 10 μl |
Procedure
2. Mix Well and incubate for 5 mins at room Tempreature or 3 min @ 37c’
3. using the colorimeter or spectrophotometer read the absorbance against blank at filter wavelenght 580 nm
4. then do the calculations based on the following formula.
Calcium = Absorbance of test / Absorbance of standard x Conc of standard
The STD Conc is 10 mg/dl or may vary depending on the reagent brand.
Normal Range
Quality Control:
1. Run two levels of control with each batch of samples.
2. Results should fall within established confidence limits.
3. Review QC results and take corrective action if needed.
Warning :
1. Hemolysis causes falsely decreased results.
M. Sulieman
mohammad@mlsgaate.com
Check glucose reagent expiration. Inspect instrument for errors. Run controls before testing samples.
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