Estimation of serum in Copper

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Estimation of serum in Copper:

Click to edit Master subtitle style Estimation of serum in Copper

Estimation of copper by flame atomic spectroscopy: :

Estimation of copper by flame atomic spectroscopy: Samples Collection and Preparation Five milliliters of venous blood transferred into sterile test tube. A llowed to clot and centrifuged at 4000rpm for 10 minutes, stored at -20ºC until analysis. Determination of Copper Samples were diluted 1:10 with 6% n- butanol solution. The level of sera copper were calculated after application of absorbencies on suitable calibration curve for each element made from standard solutions.

Modified Spectrophotometric Micromethod to Determine Serum Copper:

Modified Spectrophotometric Micromethod to Determine Serum Copper PRINCIPLE: In this method Guanidine hydrochloride was used to release ceruloplasmin bound copper and determined using bathocuproine disulphonate disodium salt (BCDS). A guanidine hydrochloride at its concentration of 4 M or greater was sufficient to completely relax and alter the tertiary structure releasing protein bound metal ions . The copper released from ceruloplasmin will form complex with copper specific chromogen BCDS, which can be estimated spectrophotometrically Company Logo

Procedure :

Procedure Calibration Curve : 100 µl of copper working standard was taken in tubes into which 200 μL of guanidine hydrochloride containing ascorbate was added . I ncubated at room temperature for 10 min, 200 μL of BCDS in tris was added, the volume is made up to 500 μL by using tris solution, vortex mixed and incubated at room temperature for 5 min Absorbance was read at 480 nm

Sample :

Sample The corrected absorbance of the sample (ST-{SB + RB}) was used to determine amount of copper by using extinction co-efficient derived from the calibration curve. Sample Test (ST), Sample Blank (SB) Reagent Blank (RB). 100 ml of serum 100 ml of serum 100 ml tris soln 200 ml of guanidine hydrochloride containing ascorbate was added . Vortex mixed 200 ml of guanidine hydrochloride containing ascorbate was added . Vortex mixed 200 ml of guanidine hydrochloride containing ascorbate was added . Vortex mixed . 200 μL BCDS in tris was added 200 μL of tris sol added . 200 μL BCDS in tris was added Absorbance was read at 480 nm after 5 min. Absorbance was read at 480 nm after 5 min. Absorbance was read at 480 nm after 5 min.

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