Determination of Triclabendazole in Cattle Plasma as Its Sulphoxide and Sulphone Metabolites by Rotating Disk Sorptive Extraction Combined With High-Performance Liquid Chromatography and Its Application to Pharmacokinetic Studies

Abstract

In this study, a new method for determination of triclabendazole (TCB) as its main metabolites, triclabendazole sulfoxide (TCB-SO) and triclabendazole sulfone (TCB-SO 2 ) in animal plasma was developed. TCB is widely used as antiparasitic in the veterinary industry. Rotating disk sorptive extraction (RDSE) was the selected sample preparation technique for extraction/clean up of the compounds from the samples followed by high performance liquid chromatography coupled to diode array detection (HPLC-DAD) for quantification.Optimization of physicochemical variables was performed using a multivariate experimental design. Following the recommendations given in the Veterinary International Conference on Harmonization guides (VICH GL02 and VICH GL49), the validation of the method was performed, given rise to improved analytical features compared with those provided by other methods based on solid phase extraction (SPE). Linearity (r > 0.99) was achieved for both compounds when calibration was performed not only in standard solutions but also in animal spiked plasma. Selectivity, defined as the response ratio between blank and analyte at the limit of quantification, was 11.8% and 3.4% for TCB-SO and TCB-SO 2 , respectively. Accuracy and precision, expressed in percentage, were always lower than -16.7% and 8.1%, respectively. Eco-efficiency was quantitatively assessed indicating that the method is an excellent green method.The proposed analytical method was applied to the determination of the pharmacokinetic of two commercial products of TCB in cattle plasma after oral administration.The good analytical performance, eco-efficiency and economy, make the method interesting for alternative use in routine laboratories.

Development and validation of a rapid selective high-throughput LC-MS/MS method for the determination of triclabendazole sulfoxide concentrations in sheep plasma and its use in bioequivalence studies

“…Various methods for the determination of triclabendazole and its metabolites in biological matrices of different species by HPLC or LC/MS, alone or simultaneously with other analytes [10][11][12][13][14][15][16] have been described in literature.…”

“…This method uses HPLC coupled with UV detection which is inferior with regards to selectivity and sensitivity compared to mass spectrometric detection. The separation is also performed on a very large HPLC column with a length of 250 mm and diameter of 4.6 mm which results in a total runtime of 13 min for each sample, compared to the much shorter runtime achieved using the smaller Gemini NX-C18, 50 3 2 mm, analytical column [13]. The method developed by Ceballos et al to measure triclabenzole and its metabolites from sheep plasma also uses protein precipitation for plasma sample purification, Fig.…”

“…The determination of drug concentrations in plasma or other biological samples through liquid chromatography coupled with mass spectrometry (LC-MS/MS) methods is now considered a technological prerequisite for highly specific and sensitive analysis. Determination of triclabendazole metabolites from different types of biological matrices in animals has been described in several studies using different analytical methods [10][11][12][13][14][15][16]. Several methods reported in literature use high performance liquid chromatography (HPLC) to determine triclabendazole and its metabolites in the plasma of different species [10,13,14].…”

“…Determination of triclabendazole metabolites from different types of biological matrices in animals has been described in several studies using different analytical methods [10][11][12][13][14][15][16]. Several methods reported in literature use high performance liquid chromatography (HPLC) to determine triclabendazole and its metabolites in the plasma of different species [10,13,14]. In recent years, several LC-MS/MS methods have been described in literature for the determination of triclabendazole and its metabolites from bovine milk [15,16], and fat, muscle, liver and kidney tissues of cattle and goat [11,12].…”

“…Analysis was performed with HPLC coupled with UV detection using acetonitrile and water as mobile phase in isocratic elution and a C18 HPLC column. Under these chromatographic conditions the total runtime is 13 min for one sample [13]. Ceballos et al developed an analytical method to measure triclabenzole and its metabolites from sheep plasma for pharmacokinetic studies [14].…”