My Chemistry Experiment
- 1. Determination of creatinine in urine of a diabetic patient using MEKCBy Robert Rezahttp://en.wikipedia.org/wiki/File:Phospholipids_aqueous_solution_structures.svg
- 2. AbstractType II diabetic patient avoiding doctor Creatinine biomarker for renal functionQuantify Creatinine in patient’s urine using MEKCHypothesis- Patient will have high levels but be with in normal limitsQuantification methods- 4 pt. calibration and internal standard all run on Groton CEResults compared to expected normal levelsAs of right now, no data
- 3. 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 injectionsWith in the past yr lost about 10 lbs Still continuing bad habits
- 4. Background-ExperimentCreatinine is present in urineBiomarker for kidney functionKidneys responsible for waste managementKidney failure more common in type II diabeticsExperiment aims to quantify creatinine in urine using MEKCCreatinine is zwitterionic between pH 4.8-9.2Normal 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
- 5. Goals and HypothesisGoal-To quantify creatinine in patient’s urine and compare to normal levelsGoal 2- quantify creatinine in patient’s urine on more than one day and compareHypothesis: patient’s creatinine content will be high (2000+) but with in normal limits
- 6. ExperimentalUrine collected morning of experiment and kept on ice till usePatient was fasting2xUrine samples centrifuged and diluted 10xSamples 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 filterHydrodynamic injection at 33.86 mbar30 mM phosphate-150 mM SDS buffer @ pH 6Sodium Dodecyl Sulfate (SDS)http://commons.wikimedia.org/wiki/File:SDS-2D-skeletal.pngLiotta,E.; Clinica Chimica Acta 409 (2009) 52-55
- 7. Experimental (continued)Groton Biosystems CE ~90 cm (end to end) x 75m capillary tube Voltage- 20kV Wavelength detection- 214 nmAll solutions run in triplicateHP water used to determine LODhttp://www.files.chem.vt.edu/chem-ed/sep/electrop/cap-el.html
- 8. Results/ ConclusionsSample data fitted to calibration curveComparison to an Internal standard, imidazoleResults compared to normal levels and patient notifiedAs of right now no data produced
- 9. Questions?