Influence of the Acetamide from Acetonitrile Hydrolysis in Acid-Contained Mobile Phase on the Ultraviolet Detection in High Performance Liquid Chromatography

The paper presents the influence of acetamide on ultraviolet detection after generation of acetonitrile by hydrolysis in the presence of trifluoroacetic acid in the HPLC mobile phase. The acetonitrile, with added varying contents of trifluoroacetic acid and water, was determined by GC-MS at various times. The results showed that the concentration of acetamide increased approximately linearly with time. Using a mixed standard sample solution and a model mobile phase of acetonitrile-water-tri-fluoroacetic acid (25:75:0.3, v/v) after 0, 24 and 48 h, the influence of the hydrolyzate acetamide on HPLC detection was investigated at the wavelength of 205-220 nm. The results showed that the limit of detection (LOD) of standard sample lost about 30 % after the mobile phase was placed 48 h. It is suggested that the acid-containing mobile phase was placed no more than 24 h for HPLC trace analysis at the wavelength of 205-220 nm.