An HPLC screening method for the detection of isonicotinic acid hydrazide in cattle milk

Summary A simple, fast, accurate and highly sensitive (5 ng/ml) method for the detection of sionicotinic acid hydrazide (isoniazid) in cattle milk is described, based upon an RP-HPLC system, equipped with a C18 column and a UV detector. Milk was deproteinized with trichloroacetic acid and treated with cinnamaldehyde, before being injected into the HPLC system. Preconcentration of samples on an SPE phenyl cartridge allows the sensitivity of the method to be increased to 0.05 ng/ml using a UV detector. The presence of isoniazid can be confirmed at concentrations as low as 0.4 ng/ml, through a diode-array UV spectrum of isoniazid hydrazone. The analytical procedure here described is routinely applied to investigate the illegal administration of isoniazid to cattle, as it provides a means of screening that can be carried out in a very short time.