The HPLC/UV method was developed, optimized and validated for the analysis of caffeine and its selected metabolites (1-methyluric acid, theobromine, paraxanthine, theophylline) in urine. The method is also used for the quantification of caffeine in various food samples.
ABSTRACT
HPLC/UV determination of caffeine (CAS 58-08-2) and its metabolites 1-methyluric acid (CAS 708-79-2), theobromine (CAS 519-41-5), paraxanthine (CAS 611-59-6) and theophylline (CAS 58-55-9), with etofylline as internal standard (CAS 519-37-9), on an ARION® Plus C18, 5 µm column (150 mm × 3.0 mm, ARI-5720-LK30). Gradient elution with A: 1% acetic acid in water and B: methanol at 1.0 ml/min and 35 °C, injection volume 65 µl, UV detection at 270 nm. All six compounds are resolved within about 6 minutes. Developed by the University of South Bohemia in České Budějovice.
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1-Methyluric acidCAS number 708-79-2
TheobromineCAS number 519-41-5
ParaxanthineCAS number 611-59-6
TheophyllineCAS number 58-55-9
Etofylline (IS)CAS number 519-37-9
CaffeineCAS number 58-08-2
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University of South Bohemia in České Budějovice, Faculty of Science
Faculty of Science of the University of South Bohemia in České Budějovice (Czech Republic). Named as the developer of the ARION(R) HPLC applications for caffeine metabolites in urine, atorvastatin photodegradation products and the degradation assessment of diltiazem.
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