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A simple HPLC method with pulsed EC detection for the analysis of creatine
Yoonsun Mo1, David Dobberpuhl, Alekha K Dash
1Department of Pharmacy Sciences, School of Pharmacy and Health Professions, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA.
Journal of Pharmaceutical and Biomedical Analysis
|July 11, 2003
Summary
A new liquid chromatography (LC) method offers sensitive detection of creatine in solutions and rat plasma. This validated method accurately quantifies creatine, distinguishing it from creatinine, and aids in studying creatine salt dissolution.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Creatine analysis is crucial for understanding its role in energy metabolism and physiological processes.
- Accurate quantification of creatine in biological matrices like plasma is essential for pharmacokinetic and metabolic studies.
- Existing analytical methods may lack the sensitivity, simplicity, or specificity required for comprehensive creatine analysis.
Purpose of the Study:
- To develop and validate a simple, sensitive, and reliable liquid chromatography (LC) method for creatine determination.
- To enable the analysis of creatine in both aqueous solutions and complex biological samples such as rat plasma.
- To establish a method capable of resolving creatine from its primary degradation product, creatinine.
Main Methods:
- Utilized a high-performance liquid chromatography (HPLC) system with a pulsed electrochemical detector (PED).
- Employed a specific mobile phase composition (water, acetonitrile, sodium acetate, sodium hydroxide) and a specialized stationary phase (polyhydroxylated glucose and sulfonated).
- Optimized chromatographic conditions including flow rate (1.0 ml/min) and column temperature (45°C) for efficient separation.
Main Results:
- Achieved excellent chromatographic separation of creatine (retention time 3.50 min) from creatinine (retention time 4.73 min).
- Demonstrated linear standard curves for creatine quantification within the range of 0-20 µg/ml.
- Reported within-day and day-to-day relative standard deviations (R.S.D.) below 10%, indicating good method precision and accuracy.
- Successfully applied the method to investigate the dissolution profiles of various creatine salts in aqueous media.
Conclusions:
- The developed LC-electrochemical detection method is simple, sensitive, and suitable for creatine quantification in aqueous solutions and rat plasma.
- The method's ability to resolve creatine from creatinine ensures accurate measurements, crucial for biological and pharmaceutical applications.
- This validated analytical approach facilitates further research into creatine's bioavailability, stability, and dissolution characteristics.