HPLC Method for Analysis mixture of Xanthines and Uric Acid BIST B+ by SIELC Technologies.
Separation type: Hydrophilic interaction liquid chromatography (HILIC)
Xanthines and uric acid are related compounds in the body and both are involved in the metabolism of purines.
Xanthines are a group of alkaloids that are widely distributed in plants, and also occur in the tissues and fluids of animals. They are known to stimulate the central nervous system and cardiac muscle, and also have diuretic effects. Some commonly known xanthines include caffeine (found in coffee, tea, and chocolate), theobromine (found in cocoa and chocolate), and theophylline (used as a drug in the treatment of respiratory diseases like asthma).
In the body, xanthines are intermediates in the degradation of adenosine monophosphate to uric acid. This metabolic pathway starts with adenosine monophosphate (AMP), which is deaminated to form inosine monophosphate (IMP). IMP is then converted into a xanthine known as hypoxanthine. Hypoxanthine is then oxidized to xanthine, and finally, xanthine is further oxidized to uric acid. Both of the oxidation steps are catalyzed by the enzyme xanthine oxidase.
Uric acid and Xanthines can be retained, analyzed, and separated using an isocratic analytical method on a BIST B+ column. The simple mobile phase for this method comprises water, acetonitrile (MeCN), and formic acid as an ionic modifier. The analytical method can be monitored with UV detection at 260 nm, an Evaporative Light Scattering Detector (ELSD), or any other evaporative detection method such as Charged Aerosol Detection (CAD) or Electrospray Ionization Mass Spectrometry (ESI-MS)
|Column||BIST B+, 4.6 x 150 mm, 10 µm, 100 A|
|Mobile Phase||MeCN – 85%|
|Buffer||FA – 0.5%|
|Flow Rate||1.0 ml/min|
|Detection||UV 260, 290 nm|
|Peak Retention Time||2.01, 3.02, 4.2, 9.09 min|
|Class of Compounds||Acid, Xanthines|
|Analyzing Compounds||Uric Acid, Caffeine, 1,3-Dimethyluric acid, Xanthine, Hypoxanthine|
Column Diameter: 4.6 mm
Column Length: 150 mm
Particle Size: 10 µm
Pore Size: 100 A