Determination of creatinine in urine of a diabetic patient using

MEKCBy Robert Reza

http://en.wikipedia.org/wiki/File:Phospholipids_aqueous_solution_structures.svg

Abstract Type II diabetic patient avoiding doctor Creatinine biomarker for renal function Quantify Creatinine in patient’s urine using

MEKC Hypothesis- Patient will have high levels but be

with in normal limits Quantification methods- 4 pt. calibration and

internal standard all run on Groton CE Results compared to expected normal levels As of right now, no data

Background-Patient Info.

Patient is my Mom (Gloria Reza) Has been diagnosed type II diabetic since

2001 Recently avoiding seeing the doctor for fear of

insulin injections With in the past yr lost about 10 lbs Still continuing bad habits

Background-Experiment Creatinine is present in urine Biomarker for kidney function Kidneys responsible for waste management Kidney failure more common in type II diabetics Experiment aims to quantify creatinine in urine using MEKC Creatinine is zwitterionic between pH 4.8-9.2 Normal levels of creatinine in urine 280-2600 ppm

Liotta,E.; Gottardo,R.; Bonizzato, L.; Pascali,J.P.; Bertaso,A.; Tagliaro, F.; Clinica Chimica Acta 409 (2009) 52-55

Goals and Hypothesis

Goal-To quantify creatinine in patient’s urine and compare to normal levels

Goal 2- quantify creatinine in patient’s urine on more than one day and compare

Hypothesis: patient’s creatinine content will be high (2000+) but with in normal limits

Experimental Urine collected morning of experiment and kept on ice till

use Patient was fasting 2xUrine samples centrifuged and diluted 10x Samples spiked with 25 l of 150ppm imidazole (Internal

Standard) 4x Creatinine calibration solutions ( 40, 160, 240, 320

ppm) All solutions syringe filtered via 2 micron filter Hydrodynamic injection at 33.86 mbar 30 mM phosphate-150 mM SDS buffer @ pH 6

http://commons.wikimedia.org/wiki/File:SDS-2D-skeletal.png Liotta,E.; Clinica Chimica Acta 409 (2009) 52-55

Sodium Dodecyl Sulfate (SDS)

Experimental (continued) Groton Biosystems CE ~90 cm (end to end) x 75m capillary tube Voltage- 20kV Wavelength detection- 214 nm All solutions run in triplicate HP water used to determine LOD

http://www.files.chem.vt.edu/chem-ed/sep/electrop/cap-el.html

Results/ Conclusions

Sample data fitted to calibration curve Comparison to an Internal standard, imidazole Results compared to normal levels and patient notified As of right now no data produced

Questions

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