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Creatinine Lab - Nick Baccari Graham Barnett Jillian Daviau...

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Nick Baccari, Graham Barnett, Jillian Daviau Section 2 Creatinine Determination Lab Objectives: The purpose of this lab was to determine the concentration of two unknown samples of creatinine by determining the absorbance of visible light at 500NM after allowing the creatinine to proceed through the Jaffe reaction. Materials and Methods: Reagents: 0.04 M Picric acid in water 0.75 N NaOH Creatinine stock solution at 1 mg/mL in 0.1 N HCI. Store at 4C. Two unknown samples containing different concentrations of creatinine. Store @ 4C. Chloroform Glassware: Graduated cylinder 10 mL Volumetric flask 100 mL Large test tubes 8 Graduated 1 mL pipet Cuvettes for spectrophotometer 8 Procedures: 1. Diluted creatinine stock solution. This was performed by the TA. 1 22. Prepare creatinine standard solution at 0.002, 0.004, 0.008, 0.012, and 0.016 mg/mL into five large test tubes labeled #1-5 using the diluted creatinine stock solution prepared. 0.002 mg/mL =0.5 mL creatinine stock solution + 3.5 mL water 0.004 mg/mL = 1.0 mL creatinine stock solution + 3.00mL water 0.008 mg/mL = 2.0 mL creatinine stock solution + 2.0 mL water 0.012 mg/mL = 3.0 mL creatinine stock solution + 1.0 mL water 0.016 mg/mL = 4.0 mL creatinine stock solution + 0 mL water 13. Add 1 mL of 0.75 NaOH to each of test tubes #1-5 before the addition of 1 mL of 0.04 M picric acid . Mix gently by inversion. Wait for 15 minutes. 1 24. Add 4 mL of water in a test tube labeled #6 followed by 1 mL of 0.75 NaOH and then 1 mL of 0.04 M picric acid. Mix gently by inversion. Wait for 15 minutes. This will be used as a blank. 3 45. Using prepared unknown solution, transfer 4 mL of each diluted unknown samples into small test tubes #7-8. Add 1 mL of 0.75 NaOH followed by 1 mL of 0.04 M picric acid to each test tube. Mix gently by inversion. Wait for 15 minutes.
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