High-performance liquid chromatographic analysis of the mycotoxin penicillic acid and its application to biological...

Penicillic acid (PA) is a carcinogenic food contaminant produced by several food-borne fungi. PA was resolved as a sharp peak by reversed-phase high-performance liquid chromatography on a small-particle (10 micrometer) column in 3-4 min by an elution system composed of acetonitrile (AN), water and glacial acetic acid (AC) with detection by ultraviolet (UV) absorbance at 254 nm. Peak height and peak area were related linearly to the amount of PA injected over a range of 5-500 ng. Reproducibility of retention time, peak height and peak area was demonstrated. The lower detection limit was 5 ng in two elution systems [AN-H2O-AC(45:55:2), flow-rate (F) 1 ml/min; AN-H2O-AC(40:60:2), F = 1.2 ml/min] and 10 ng in a third system [AN-H2O-AC(25:75:2), F = 1.6 ml/min]. Based on the sensitivity and separation of PA from (40:60:2), F = 12 ml/min; urine, AN-H2O-AC (25:75:2), F = 1.6 ml/min. Good recovery (89-98%) over a range of 1-50 microgram/ml of PA was obtained from PA-spiked plasma samples treated first with 25% HPO3 followed by extraction with chloroform. A single peak detected either by UV absorbance or by radioactivity was obtained when plasma samples spiked with [14C]PA were extracted. Good recovery (92-105%) of PA also was obtained from spiked urine and bile samples.