An ion-pairing reversed-phase high-performance liquid chromatography (HPLC) method with diode array detection at 280 nm was developed to determine pyrimethamine concentrations in feed for laying hens. Pyrimethamine was extracted with a mixture of 5% isobutanol and 95% benzene, and the extract was cleaned up on an alumina column. The drug was eluted from an Intersil ODS-3V column (250 by 4.6 mm) with a mixture of 25% acetonitrile and 75% water (vol/vol) containing 0.01 M tetramethylammonium chloride as an ion-pairing agent and adjusted with acetic acid to pH 3.5. The flow rate was 1.0 ml/min. Mean recovery of pyrimethamine from supplemented feeds at concentrations of 2, 4, and 5 μg/g of feed were 100.5, 103.5, and 100.8%, respectively. Precision within a day ranged from 4.3 to 7.0% for the three concentrations, and day-to-day precision was 5.3% for feed supplemented at a concentration of 4 μg/g. No chromatographic interference was detected from other 2,4-diaminopyrimidine compounds or other major drugs used in poultry.
Development of a Method for the Determination of Pyrimethamine Concentrations in Feeds by Ion-Pairing High-Performance Liquid Chromatography
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P. S. GONG, S. L. JENG, Y. F. HSU, C. C. LIN, S. Y. LIN; Development of a Method for the Determination of Pyrimethamine Concentrations in Feeds by Ion-Pairing High-Performance Liquid Chromatography. J Food Prot 1 April 2002; 65 (4): 688–691. doi: https://doi.org/10.4315/0362-028X-65.4.688
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