Separation type: Hydrophilic interaction liquid chromatography (HILIC)

Uric acid is the final product of purine metabolism and is commonly found in urine. Using SIELC’s newly introduced BIST™ method, this purine metabolite can be retained and separated from urine and plasma 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, Uric Acid can be separated, retained, and UV detected at 260 and 290 nm.

High Performance Liquid Chromatography (HPLC) Method for Analysis of Uric Acid in Urine and Human Serum Samples

Condition

Column	BIST™ B+, 4.6×150 mm, 5µm, 100A
Mobile Phase	MeCN – 85%
Buffer	Formic Acid – 0.5%
Flow Rate	1.0 ml/min
Detection	UV 290 nm
Retention time	9.09 min

Description

Class of Compounds	Acid
Analyzing Compounds	Uric Acid