Pipette-tip solid-phase extraction using polypyrrole as efficient adsorbent for extraction of avermectins and milbemycins in milk
In this work, we developed a HPLC method for the multidetermination of avermectins (AVM) (abamectin—ABA 1b and ABA 1a, eprinomectin—EPR, and ivermectin—IVM) and milbemycins (moxidectin—MOX) in milk samples using polypyrrole (PPy) as adsorbent material in pipette-tip solid-phase extraction (PT–PPy–SPE). PPy was characterized by scanning electron microscopy, thermogravimetric analysis, Fourier transform infrared spectroscopy, and X-ray diffraction and the data agreed with the literature. The sample preparation included the clean-up of the milk by protein precipitation (PP) with acetonitrile and extraction of the analytes by PT–PPy–SPE. The chromatographic method was developed in reverse phase and isocratic mode with flow rate at 1.2 mL min−1 and ultraviolet detection at 250 nm. The mobile phase composition was acetonitrile:methanol:water (55:25:20, v/v/v). The studied parameters and the optimized conditions for the sample preparation were washing solvent (300 μL water), volume and type of eluent (500 μL methanol), volume and pH of sample (1 mL and pH 10), amount of adsorbent material (50 mg PPy), and without addition of salt (NaCl). The method was linear over the concentration range from 20 to 3000 ng mL−1 with coefficients of correlation (r) ≥ 0.99 for all analytes and recoveries around 100%. The method developed and validated was used for the analyses of real milk samples from cow treated with Ivomec® (IVM 3.5%), in which were found 21.51 ± 2.94 ng mL−1 of IVM. Finally, the results proved that PT–PPy–SPE coupled to HPLC–UV was economical, simple, and easy-to-perform technique.
KeywordsAvermectins Milbemycyns Polypyrrole Pipette-tip solid-phase extraction Milk
The authors would like to thank the Brazilian agencies CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), FAPES (Fundação de Amparo à Pesquisa e Inovação do Espírito Santo), and FAPEMIG (Fundação de Amparo à Pesquisa do Estado de Minas Gerais) for financial support. This study is also part of the project involving the Rede Mineira de Química (RQ-MG) supported by FAPEMIG (Project: REDE-113/10; Project: CEX-RED-0010-14).
Compliance with ethical standards
All studies involving animals are reported in accordance with the ARRIVE guidelines and approved by the institutional ethics board committee.
Conflict of interest
The authors declare that they have no conflict of interest.
- 5.Council Directive 96/23/Ec. Measures to monitor certain substances and residues thereof in live animals and animal products and repealing. Directives 85/358/EEC and 86/469/EEC and Decisions 89/187/EEC and 91/664/EEC. 2012;127:10–31.Google Scholar
- 6.Brasil, Ministério da Agricultura, Pecuária e Abastecimento, Secretaria de Defesa Agropecuária, Instrução Normativa n° 46 22/11/2016.Google Scholar
- 9.Gomes LVC, Lopes WDZ, Cruz BC, Teixeira WF, Felippelli G, Maciel WG, et al. Acaricidal effects of fluazuron (2.5 mg/kg) and a combination of fluazuron (1.6 mg/kg) + ivermectin (0.63 mg/kg), administered at different routes, against Rhipicephalus (Boophilus) microplus-parasitizing cattle. Exp Parasitol. 2015;153:22–8.CrossRefPubMedGoogle Scholar
- 19.Boscher A, Guignard C, Pellet T, Hoffmann L, Bohn T. Development of a multi-class method for the quantification of veterinary drug residues in feedingstuffs by liquid chromatography–tandem mass spectrometry. J Chromatogr A. 2010;(41):6394–404.Google Scholar
- 20.Kaufmann P. Multi-residue quantification of veterinary drugs in milk with a novel extraction and clean up technique: salting out supported liquid extraction (SOSLE). Anal Chem. 2014;820:56–68.Google Scholar
- 26.Durden DA. Positive and negative electrospray LC–MS–MS methods for quantitation of the antiparasitic endectocide drugs, abamectin, doramectin, emamectin, eprinomectin, ivermectin, moxidectin and selamectin in milk. J Chromatogr A. 2007;850:134–57.Google Scholar
- 29.Andrade RT, da Silva RCS, Pereira AC, Borges KB. Self-assembly pipette tip-based cigarette filters for micro-solid phase extraction of ketoconazole cis-enantiomers in urine samples followed by high performance liquid chromatography/diode array detection. Anal Methods. 2015;7:7270–80.CrossRefGoogle Scholar
- 30.da Silva RCS, Mano V, Pereira AC, de Figueiredo EC, Borges KB. Development of pipette tip-based on molecularly imprinted polymer micro-solid phase extraction for selective enantioselective determination of (−)-(2S,4R) and (+)-(2R,4S) ketoconazole in human urine samples prior to HPLC-DAD. Anal Methods. 2016;8:4075–85.CrossRefGoogle Scholar
- 33.Borges KB, de Figueiredo EC, Queiroz MEC (2015) Preparo de amostras para análise de compostos orgânicos, first ed., LTC−Livros Técnicos Científicos Editora Ltda, Rio de Janeiro.Google Scholar
- 38.Meng J, Bu J, Deng C, Zhang X. Preparation of polypyrrole-coated magnetic particles for micro solid-phase extraction of phthalates in water by gas chromatography–mass spectrometry analysis. J Chromatogr A. 2011;(12):1585–91.Google Scholar
- 41.VICH GL 49, Studies to evaluate the metabolism and residue kinetics of veterinary drugs in food-producing animals: validation of analytical methods used in residue depletion studies. http://www.vichsec.org/guidelines/pharmaceuticals/pharma-safety/metabolism-and-residue-kinetics.html, 2015 (accessed 08 June 2017).
- 42.Turnipseed S, Roybal J, Andersen W, Kuck L. Analysis of avermectin and moxidectin residues in milk by liquid chromatography–tandem mass spectrometry using an atmospheric pressure chemical ionization/atmospheric pressure photo ionization source. Anal Chem. 2005;529:159–65.Google Scholar
- 46.Frenich AG, Aguilera MDM, Vidal JLM, Romero-González R. Comparison of several extraction techniques for multiclass analysis of veterinary drugs in eggs using ultra-high pressure liquid chromatography–tandem mass spectrometry. Anal Chem. 2010;(2):150–60.Google Scholar
- 47.Priscila MS, Maia B, Rezende DF, Annibal D, Netto FP, Marques FC. An alternative derivatization reaction to the determination of doramectin in bovine milk using spectrofluorimetry. Spectrochim Acta A Mol Biomol Spectrosc. 2015;100:127–30.Google Scholar
- 49.Imperiale FA, Busetti MR, Suarez VH, Lanusse CE. Milk excretion of ivermectin and moxidectin in dairy sheep: assessment of drug residues during cheese elaboration and ripening period. J Agric Food Chem. 2009;20:6205–11.Google Scholar
- 50.Campillo N, Viñas P, Férez-Melgarejo G, Hernández-Córdoba M. Dispersive liquid–liquid microextraction for the determination of macrocyclic lactones in milk by liquid chromatography with diode array detection and atmospheric pressure chemical ionization ion-trap tandem mass spectrometry. J Chromatogr A. 2013;1282:20–6.CrossRefPubMedGoogle Scholar