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High Performance Liquid Chromatography Determination of Urinary Hippuric Acid and Benzoic Acid as Indices for Glue Sniffer Urine

Authors: Abdul Rahim Yacob, Mohamad Raizul Zinalibdin


A simple method for the simultaneous determination of hippuric acid and benzoic acid in urine using reversed-phase high performance liquid chromatography was described. Chromatography was performed on a Nova-Pak C18 (3.9 x 150 mm) column with a mobile phase of mixed solution methanol: water: acetic acid (20:80:0.2) and UV detection at 254 nm. The calibration curve was linear within concentration range at 0.125 to 6.0 mg/ml of hippuric acid and benzoic acid. The recovery, accuracy and coefficient variance of hippuric acid were 104.54%, 0.2% and 0.2% respectively and for benzoic acid were 98.48%, 1.25% and 0.60% respectively. The detection limit of this method was 0.01ng/l for hippuric acid and 0.06ng/l for benzoic acid. This method has been applied to the analysis of urine samples from the suspected of toluene abuser or glue sniffer among secondary school students at Johor Bahru.

Keywords: Glue sniffer, High Performance LiquidChromatography, Hippuric Acid, Toluene, Urine.

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