Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees

Autores
Reinaud, Daniel; Perez, Vanina Andrea; Vidondo, Patricio Federico; del Hoyo, Marcelo Luis; Lanusse, Carlos Edmundo; Alvarez, Luis Ignacio; Moreno Torrejon, Laura
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Introduction/Objective: Amitraz (AMT) is a formamidine ectoparasiticide used in veterinary medicine to control ticks, lice and mites. It is available for topical use as spray, dip, or pour on in dogs, swine and cattle. Argentina is one of the major honey bee producer countries. The mite Varroa destructor is a pathogenic parasite, which seriously affect honey production by Apis mellifera. It feeds on the haemolymph of brood and adult honey bees, contributing to the spread of viral and bacterial infections. AMT has been approved to use as a sustained-released strip to control varroasis. The goal of the present work was to develop and validate an HPLC analytical methodology to quantify AMT after its recovery from PVC fortified strips (AMIVAR 500®, APILAB S.R.L.). Materials and Methods: AMT quantification was performed by HPLC with UV detection (288 nm). An Agilent Extend-C18 reversed-phase column (4.6 × 250 mm, 5 µm) fitted with a guard column (4.6 × 12.5 mm, 5 µm) was used for separation. Elution was carried out at an isocratic flow (1 ml per min) using a mobile phase composed by 20 mM triethylamine: acetonitrile (10:90). AMT was extracted from PVC strip samples with methylene chloride for 18 h at 20 ± 2°C. The method underwent an exhaustive validation process following internationally recognized standards (CVMP, VICH, 1998). Considering the purpose of the developed analytical method, the following validation parameters were determined: specificity, linearity, range, precision (repeatability, intermediate precision) and recovery. Results and Conclusions: The AMT retention time was a 7.5 min. No interferences from endogenous compounds of the PVC strip sample were observed. The ratio “AMT standard signal/blank noise signal” was much higher than that required to determine the measurable concentration range (20–30 µg ml−1). The calibration parameters achieved the established acceptance criteria, repeatability of the triplicates (intraday repeatability) with a CV ≤ 2% and correlation coefficient ≥ 0.99. The repeatability (CV) and intermediate precision were ≤ 2% or ≤ 4%, respectively. The recovery was 104% so it was proposed not to correct for recovery in the routine analysis. The obtained validation parameters confirm the linearity of the model, offering a good fit for the proposed working range. Thus, the developed analytical methodology complies with the validation criteria required for AMT quantification in PVC strips. The analytical data shown here is a step forward to assure the correct use of AMT to control Varroa mite in honey bee production.
Fil: Reinaud, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Perez, Vanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Vidondo, Patricio Federico. Apilab Sociedad de Responsabilidad Limitada.; Argentina
Fil: del Hoyo, Marcelo Luis. Apilab Sociedad de Responsabilidad Limitada.; Argentina
Fil: Lanusse, Carlos Edmundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Alvarez, Luis Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Moreno Torrejon, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
14 th International Congress of the European Association for Veterinary Pharmacology and Toxicology
Wroclaw
Polonia
European Association for Veterinary Pharmacology and Toxicology
Materia
Development and validation
Analytical technique
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/167665
Acceder
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spelling Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey beesReinaud, DanielPerez, Vanina AndreaVidondo, Patricio Federicodel Hoyo, Marcelo LuisLanusse, Carlos EdmundoAlvarez, Luis IgnacioMoreno Torrejon, LauraDevelopment and validationAnalytical techniquehttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Introduction/Objective: Amitraz (AMT) is a formamidine ectoparasiticide used in veterinary medicine to control ticks, lice and mites. It is available for topical use as spray, dip, or pour on in dogs, swine and cattle. Argentina is one of the major honey bee producer countries. The mite Varroa destructor is a pathogenic parasite, which seriously affect honey production by Apis mellifera. It feeds on the haemolymph of brood and adult honey bees, contributing to the spread of viral and bacterial infections. AMT has been approved to use as a sustained-released strip to control varroasis. The goal of the present work was to develop and validate an HPLC analytical methodology to quantify AMT after its recovery from PVC fortified strips (AMIVAR 500®, APILAB S.R.L.). Materials and Methods: AMT quantification was performed by HPLC with UV detection (288 nm). An Agilent Extend-C18 reversed-phase column (4.6 × 250 mm, 5 µm) fitted with a guard column (4.6 × 12.5 mm, 5 µm) was used for separation. Elution was carried out at an isocratic flow (1 ml per min) using a mobile phase composed by 20 mM triethylamine: acetonitrile (10:90). AMT was extracted from PVC strip samples with methylene chloride for 18 h at 20 ± 2°C. The method underwent an exhaustive validation process following internationally recognized standards (CVMP, VICH, 1998). Considering the purpose of the developed analytical method, the following validation parameters were determined: specificity, linearity, range, precision (repeatability, intermediate precision) and recovery. Results and Conclusions: The AMT retention time was a 7.5 min. No interferences from endogenous compounds of the PVC strip sample were observed. The ratio “AMT standard signal/blank noise signal” was much higher than that required to determine the measurable concentration range (20–30 µg ml−1). The calibration parameters achieved the established acceptance criteria, repeatability of the triplicates (intraday repeatability) with a CV ≤ 2% and correlation coefficient ≥ 0.99. The repeatability (CV) and intermediate precision were ≤ 2% or ≤ 4%, respectively. The recovery was 104% so it was proposed not to correct for recovery in the routine analysis. The obtained validation parameters confirm the linearity of the model, offering a good fit for the proposed working range. Thus, the developed analytical methodology complies with the validation criteria required for AMT quantification in PVC strips. The analytical data shown here is a step forward to assure the correct use of AMT to control Varroa mite in honey bee production.Fil: Reinaud, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Perez, Vanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Vidondo, Patricio Federico. Apilab Sociedad de Responsabilidad Limitada.; ArgentinaFil: del Hoyo, Marcelo Luis. Apilab Sociedad de Responsabilidad Limitada.; ArgentinaFil: Lanusse, Carlos Edmundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Alvarez, Luis Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; ArgentinaFil: Moreno Torrejon, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina14 th International Congress of the European Association for Veterinary Pharmacology and ToxicologyWroclawPoloniaEuropean Association for Veterinary Pharmacology and ToxicologyWiley2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciatext/plainapplication/vnd.openxmlformats-officedocument.wordprocessingml.documentapplication/pdfhttp://hdl.handle.net/11336/167665Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees; 14 th International Congress of the European Association for Veterinary Pharmacology and Toxicology; Wroclaw; Polonia; 2018; 119-1200140-77831365-2885CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/10.1111/jvp.12654info:eu-repo/semantics/altIdentifier/doi/10.1111/jvp.12654Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:07:02Zoai:ri.conicet.gov.ar:11336/167665instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:07:02.5CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
title Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
spellingShingle Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
Reinaud, Daniel
Development and validation
Analytical technique
title_short Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
title_full Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
title_fullStr Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
title_full_unstemmed Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
title_sort Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees
dc.creator.none.fl_str_mv Reinaud, Daniel
Perez, Vanina Andrea
Vidondo, Patricio Federico
del Hoyo, Marcelo Luis
Lanusse, Carlos Edmundo
Alvarez, Luis Ignacio
Moreno Torrejon, Laura
author Reinaud, Daniel
author_facet Reinaud, Daniel
Perez, Vanina Andrea
Vidondo, Patricio Federico
del Hoyo, Marcelo Luis
Lanusse, Carlos Edmundo
Alvarez, Luis Ignacio
Moreno Torrejon, Laura
author_role author
author2 Perez, Vanina Andrea
Vidondo, Patricio Federico
del Hoyo, Marcelo Luis
Lanusse, Carlos Edmundo
Alvarez, Luis Ignacio
Moreno Torrejon, Laura
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Development and validation
Analytical technique
topic Development and validation
Analytical technique
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Introduction/Objective: Amitraz (AMT) is a formamidine ectoparasiticide used in veterinary medicine to control ticks, lice and mites. It is available for topical use as spray, dip, or pour on in dogs, swine and cattle. Argentina is one of the major honey bee producer countries. The mite Varroa destructor is a pathogenic parasite, which seriously affect honey production by Apis mellifera. It feeds on the haemolymph of brood and adult honey bees, contributing to the spread of viral and bacterial infections. AMT has been approved to use as a sustained-released strip to control varroasis. The goal of the present work was to develop and validate an HPLC analytical methodology to quantify AMT after its recovery from PVC fortified strips (AMIVAR 500®, APILAB S.R.L.). Materials and Methods: AMT quantification was performed by HPLC with UV detection (288 nm). An Agilent Extend-C18 reversed-phase column (4.6 × 250 mm, 5 µm) fitted with a guard column (4.6 × 12.5 mm, 5 µm) was used for separation. Elution was carried out at an isocratic flow (1 ml per min) using a mobile phase composed by 20 mM triethylamine: acetonitrile (10:90). AMT was extracted from PVC strip samples with methylene chloride for 18 h at 20 ± 2°C. The method underwent an exhaustive validation process following internationally recognized standards (CVMP, VICH, 1998). Considering the purpose of the developed analytical method, the following validation parameters were determined: specificity, linearity, range, precision (repeatability, intermediate precision) and recovery. Results and Conclusions: The AMT retention time was a 7.5 min. No interferences from endogenous compounds of the PVC strip sample were observed. The ratio “AMT standard signal/blank noise signal” was much higher than that required to determine the measurable concentration range (20–30 µg ml−1). The calibration parameters achieved the established acceptance criteria, repeatability of the triplicates (intraday repeatability) with a CV ≤ 2% and correlation coefficient ≥ 0.99. The repeatability (CV) and intermediate precision were ≤ 2% or ≤ 4%, respectively. The recovery was 104% so it was proposed not to correct for recovery in the routine analysis. The obtained validation parameters confirm the linearity of the model, offering a good fit for the proposed working range. Thus, the developed analytical methodology complies with the validation criteria required for AMT quantification in PVC strips. The analytical data shown here is a step forward to assure the correct use of AMT to control Varroa mite in honey bee production.
Fil: Reinaud, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Perez, Vanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Vidondo, Patricio Federico. Apilab Sociedad de Responsabilidad Limitada.; Argentina
Fil: del Hoyo, Marcelo Luis. Apilab Sociedad de Responsabilidad Limitada.; Argentina
Fil: Lanusse, Carlos Edmundo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Alvarez, Luis Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
Fil: Moreno Torrejon, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina
14 th International Congress of the European Association for Veterinary Pharmacology and Toxicology
Wroclaw
Polonia
European Association for Veterinary Pharmacology and Toxicology
description Introduction/Objective: Amitraz (AMT) is a formamidine ectoparasiticide used in veterinary medicine to control ticks, lice and mites. It is available for topical use as spray, dip, or pour on in dogs, swine and cattle. Argentina is one of the major honey bee producer countries. The mite Varroa destructor is a pathogenic parasite, which seriously affect honey production by Apis mellifera. It feeds on the haemolymph of brood and adult honey bees, contributing to the spread of viral and bacterial infections. AMT has been approved to use as a sustained-released strip to control varroasis. The goal of the present work was to develop and validate an HPLC analytical methodology to quantify AMT after its recovery from PVC fortified strips (AMIVAR 500®, APILAB S.R.L.). Materials and Methods: AMT quantification was performed by HPLC with UV detection (288 nm). An Agilent Extend-C18 reversed-phase column (4.6 × 250 mm, 5 µm) fitted with a guard column (4.6 × 12.5 mm, 5 µm) was used for separation. Elution was carried out at an isocratic flow (1 ml per min) using a mobile phase composed by 20 mM triethylamine: acetonitrile (10:90). AMT was extracted from PVC strip samples with methylene chloride for 18 h at 20 ± 2°C. The method underwent an exhaustive validation process following internationally recognized standards (CVMP, VICH, 1998). Considering the purpose of the developed analytical method, the following validation parameters were determined: specificity, linearity, range, precision (repeatability, intermediate precision) and recovery. Results and Conclusions: The AMT retention time was a 7.5 min. No interferences from endogenous compounds of the PVC strip sample were observed. The ratio “AMT standard signal/blank noise signal” was much higher than that required to determine the measurable concentration range (20–30 µg ml−1). The calibration parameters achieved the established acceptance criteria, repeatability of the triplicates (intraday repeatability) with a CV ≤ 2% and correlation coefficient ≥ 0.99. The repeatability (CV) and intermediate precision were ≤ 2% or ≤ 4%, respectively. The recovery was 104% so it was proposed not to correct for recovery in the routine analysis. The obtained validation parameters confirm the linearity of the model, offering a good fit for the proposed working range. Thus, the developed analytical methodology complies with the validation criteria required for AMT quantification in PVC strips. The analytical data shown here is a step forward to assure the correct use of AMT to control Varroa mite in honey bee production.
publishDate 2018
dc.date.none.fl_str_mv 2018
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info:eu-repo/semantics/conferenceObject
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status_str publishedVersion
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dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/167665
Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees; 14 th International Congress of the European Association for Veterinary Pharmacology and Toxicology; Wroclaw; Polonia; 2018; 119-120
0140-7783
1365-2885
CONICET Digital
CONICET
url http://hdl.handle.net/11336/167665
identifier_str_mv Development and validation of an analytical technique to quantify amitraz in a PVC strip formulation used in Varroa mite control in honey bees; 14 th International Congress of the European Association for Veterinary Pharmacology and Toxicology; Wroclaw; Polonia; 2018; 119-120
0140-7783
1365-2885
CONICET Digital
CONICET
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