Determination of halofuginone in bovine plasma by competing‐ion high performance liquid chromatography after solid phase extraction

Abstract

A high performance liquid chromatography (HPLC) method for the determination of the anticoccidial and antitheilerial drug halofuginone in bovine plasma was developed. Samples were diluted with acetic acid (10%, v/v) and cleaned up on a Bond Elut C8 column. The analyte was eluted from the extraction column and chromatographed by reversed-phase HPLC using decylamine as a competing-ion reagent. Detection was by UV at 243 nm. Recovery from plasma was 75%, and within-day and between-day coefficients of variation were 5.23 and 6.35% respectively. The specificity and sensitivity of this method (limit of detection in plasma, 1 ng/mL) were sufficiently high to enable us to characterize the time course of the drug in plasma after oral administration of therapeutic doses to cattle.

HPLC Method for Quantification of Halofuginone in Human Ureter: Ex- Vivo Application

“…The absorption maxima of HFG are at 243, 285, 315, and 325 nm ( Figure 2). In the literature, most authors have used a wavelength of 243, which allows a maximum absorption of the molecule and is in agreement with the absorbance of the IS, which has an absorption maximum at 250 nm [9,14,16].…”

“…The choice of the composition of the mobile phase was directed by the literature. Indeed, many authors use a mixture of acetate buffer and acetonitrile [9,[11][12][13][14]16,29]. The tests were carried out on solutions of 20μg/ml HFG in a solution of ammonium acetate 0.25 M. Preliminary tests were performed by varying several parameters: the elution system of the mobile phase, the proportion of organic solvent, the concentration of the buffer, and the pH of mobile phase.…”

“…Several methods are available for determining HFG in feeds and tissues [9][10][11][12][13][14][15][16]. In fact, in broiler production, HFG is used during almost all the breeding periods except for pre-slaughter withdrawal.…”

Liquid chromatographic tandem mass spectrometric determination of five coccidiostats in poultry eggs and feed

“…Out of all the papers which have been published on the determination of halofuginone in different matrices, only few of them used LC-MS [4,5,[37][38][39]. Other techniques used were LC with UV detection [40][41][42][43][44][45][46][47], gas-liquid chromatography [48] and capillary isotachophoresis combined with capillary zone electrophoresis [49]. Most papers describing a method for the detection of nicarbazin in different matrices, use LC with UV detection [50][51][52][53][54][55][56][57][58][59][60][61][62][63][64].…”

Liquid chromatography-electrospray tandem mass spectrometric assay suitable for quantitation of halofuginone in plasma

“…Although LC-MS/MS assays have been developed for quantitating halofuginone in chicken liver and eggs [24,25], they are not well-suited for quantitating halofuginone in plasma because one assay does not employ an internal standard and both assays involve labor-intensive extraction techniques and large sample volumes. In addition, older GLC [26] and HPLC assays [27][28][29][30][31][32][33], including one previously developed in our laboratory [27], for quantitation of halofuginone in animal feed or biological matrices are not sufficiently sensitive to quantitate the plasma concentrations of halofuginone anticipated to be produced by the clinical doses of halofuginone being used topically or being proposed for evaluation as systemic therapy. As a result, an HPLC-electrospray tandem mass spectrometric (LC-MS/MS) assay has been developed to quantitate anticipated clinically relevant concentrations of halofuginone in plasma.…”