Creatinine Manual Method*

Background Information
Please refer to **Package Insert for specific information
Calculate the 2 SD ranges (below) for QC (please enter your ranges on your Lab Worksheet)
Enter all your Absorbance values and calculated values on the Lab Worksheet
Hand in your worksheet
Procedure
1. Please work in pairs. Watch your time carefully. Please be sure to stagger your time at
the spectrophotometer by arranging with those sharing the spectrophotometer as time is
critical in this assay.
2. This procedure calls for making a Working reagent by combining 5 volumes of Reagent 1
to 1 volume of Reagent 2. We will prepare this for you.
3. Pipette 3.0 mL of working reagent into 5 test tubes marked as BLK, STD, PAT, CTL1,
CTL2.
4. Add 200L of sample (e.g., standard, patient, controls) into each test tube. Mix and
measure the absorbance.
5. This assay is a bit different from assays we have already completed, we will read the
initial absorbance (A1 ) wait 60 seconds and read the absorbance again (A2 ) .
6. Read the absorbance at 510nm after setting the instrument to zero ABS with the
working reagent (BLK).
7. Calculate the change in absorbance by subtracting the first absorbance reading from the
second reading. A2A1.
Calculations
A2-A1 of unknown
Creatinine = _____________________________ X 2.5 mg/dL
A2-A1 of standard
Expected ranges from manufacturer: 0.40 1.40 g/dL
Linearity of assay: 0.1 25.0 mg/dL
Controls
Mea
n

Range
1SD

Range 2SD

Control
1
Control
2

Reference Range
Safety
*Please be careful as the PICRIC acid is a strong agent. Avoid contact with skin or your clothes.
Wipe all spills, use proper personal protection equipment for all labs.

Additional Questions
1. What is the classic name of the procedure performed above for measuring Creatine?

2. Describe the primary clinical reasons for measuring serum Creatinine.

3. Describe any additional testing you would recommend in a patient with an abnormal
Creatinine.

4. Describe the patient values obtained in relation to the reference range.