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Module 12: Quantitative Analysis of Urine Creatinine: Name: Group No.: 6

1. The document describes a procedure to quantify creatinine in urine using a spectrophotometer. Various standard creatinine solutions and a urine sample were prepared in test tubes and their absorbances measured. 2. A calibration curve was constructed from the standard solutions, showing a linear relationship between concentration and absorbance. This was used to calculate the creatinine concentration in the urine sample. 3. Creatinine is produced from muscle metabolism and excreted in urine, making its quantification useful for estimating lean body mass and the completeness of 24-hour urine collections.

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Ma. Lara Micaela Legaspi
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460 views3 pages

Module 12: Quantitative Analysis of Urine Creatinine: Name: Group No.: 6

1. The document describes a procedure to quantify creatinine in urine using a spectrophotometer. Various standard creatinine solutions and a urine sample were prepared in test tubes and their absorbances measured. 2. A calibration curve was constructed from the standard solutions, showing a linear relationship between concentration and absorbance. This was used to calculate the creatinine concentration in the urine sample. 3. Creatinine is produced from muscle metabolism and excreted in urine, making its quantification useful for estimating lean body mass and the completeness of 24-hour urine collections.

Uploaded by

Ma. Lara Micaela Legaspi
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
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NAME: GROUP NO.

: 6
BOLIVAR, Leonilyn SECTION: BSN1-A
LEGASPI, Ma. Lara Micaela INSTRUCTOR: Ms. Leslie Sampollo
OSUMO, Betiara Jazel
SCORE:
PELINGON, Robert Kris
PIO, Krea Joy
YSULAN, Cyky Mae DATE: January 19, 2021

Module 12: Quantitative Analysis of Urine Creatinine

I. MATERIALS

Micropipette tips 24 mL, 10% Sodium Hydroxide

Glass pipette 100 mL Distilled Water

Test tube Urine

Vernier spectrophotometer 95 mL, 1% Picric Acid Solution

Standard creatinine solution

II. PROCEDURES

1. Prepare 5 test tubes and add the following:


Test tube 1
 unknown sample
 50 uL urine sample
Test tube 2
 standard solution (0.4 mg/mL)
 10 uL std solution + 40 uL dH20
Test tube 3
 Standard solution (1.0 mg/mL)
 25 uL std solution + 25 uL dH20
Test tube 4
 Standard solution (2.0 mg/mL)
 50 uL std solution
Test tube 5
 blank
 50 uL dH20

2. Add two mL of picric acid to each of the test tubes.


3. Followed by the addition of 150 uL NaOh to each test tubes.
4. Mix and let it stand for 15 mins.
5. Dilute each solution to 10 mL (add 7.80 mL dH20).
6. For blank solution, the creatinine standard solutions concentrations:
0.4 mg/mL
1.0 mg/mL
2.0 mg/mL
7. Transfer the urine sample to a cuvette and read the absorbance of the solution in
a UV-vis spectrophotometer at wavelength 520 nm.

III. RESULTS AND DISCUSSION

Sample Concentration (mg/mL) Absorbance

blank 0 0.012

Std soln 1 0.4 0. 225

Std soln 2 1.0 0. 571

Std soln 3 2.0 1. 055

urine sample -0.040 0.082

equation of the line: y= 0.5231x + 0.0091


creatinine coeffecient: ____________ (show your solution below)
Attach the screenshot of the calibration curve computed in the excel file.
Discuss the spectrophotometer, Beer-Lamberts law
show the calculation for the creatinine in the urine sample
Post-Lab Questions
Answer the following questions briefly.
1) What are the sources of creatinine excreted in the urine?
2) Why is creatinine excretion used to estimate the completeness of a 24-hour urine
collection and as a basis for normalizing urinary concentrations of various metabolites
and estimating the lean body mass of individuals?
3) How is creatinine excretion affected by dietary meat intake?
4) What are the other uses of measuring creatinine excretion ?

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