HPLC Method for Analysis of Xanthines and Uric Acid on BIST B+ Column by SIELC Technologies

Separation type: Hydrophilic interaction liquid chromatography (HILIC)

HPLC Method for Analysis of Xantines and Uric Acid on BIST B+ Column by SIELC Technologies
HPLC Method for Analysis ofXantines and Uric Acid on BIST B+ by SIELC Technologies.

Xanthine is a purine base that naturally occurs in the human body, and is a precursor for many other stimulants, medications, and biological compounds, including caffeine and uric acid. Caffeine is one of the most well-known stimulants due to its presence in popular drinks such as coffee, tea, and soda. 1,3-Dimethyluric acid is another xanthine-derived metabolite commonly found in urine after the metabolization of xanthine derivatives. Hypoxanthine is an important purine-derivative that serves as a key nitrogen source for some types of bacteria and other cells. Uric acid is the final product of purine metabolism and is commonly found in urine. Using SIELC’s newly introduced BIST™ method, these purine-derivatives and metabolites can be retained and separated on a positively-charged anion-exchange BIST™ B+ column. 

There are two keys to this retention method: 1) a multi-charged, negative buffer, such as Sulfuric acid (H2SO4), which acts as a bridge, linking the positively-charged analytes to the positively-charged column surface and 2) a mobile phase consisting of a majority of organic solvent (such as MeCN) to minimize the formation of a solvation layer around the charged analytes. Using this new and unique analysis method, Xanthine and Uric Acid can be separated, retained, and UV detected at 260 and 290 nm.

High Performance Liquid Chromatography (HPLC) Method for Analysis of Xanthines and Uric Acid

ColumnBIST™ B+, 4.6×150 mm, 5µm, 100A
Mobile PhaseMeCN – 85%
BufferFormic Acid – 0.5%
Flow Rate1.0 ml/min
DetectionUV 290 nm
Retention time9.09 min
Class of CompoundsAcid
Analyzing CompoundsUric Acid