title: Laboratory validation of an LC-MS/MS method for the detection of ractopamine, clenbuterol and salbutamol in bovine and swine muscle at sub-μg kg-1regulatory limits.
abstract: Ractopamine (RAC), is a β-adrenergic agonist increasingly used in the swine and cattle industry. This compound redirects nutrients to favour leanness rather than fat deposition, improves growth and carcass traits gaining higher economic benefit to producers. Countries around the world are split over whether to allow the use of RAC in meat production. Clenbuterol (CLB) and salbutamol (SLB) are anillinic and phenolic β-agonists, respectively, with the same capacity of producing economic benefits for the meat sector. However, they are prohibited because of the potentially adverse reactions they can cause in consumers. The three β-agonist compounds have been included in the Brazilian National Regulatory Survey and consequentially there is an eminent need for reliable methods capable of detecting those substances at the same time and reduce analytical costs. Therefore, an LC-MS/MS method for the simultaneous determination of residual RAC, CLB and SAL in swine and cattle muscle was developed and validated with quantification levels respecting the action levels established for Brazil which are 0.1, 0.2 and 5 µg kg-1for RAC, CLB and SAL, respectively. Samples were quantified using RAC-d5, CLB-d9and SLB-d6as internal standards. The validation was performed according to European Union Decision 2002/657, which includes criteria (CCα, CCβ, recovery, repeatability, reproducibility and calibration curve). The method meets the Brazilian regulatory requirement that establishes criteria and procedures for the determination of parameters such as CCα, CCβ, precision and recovery. CCα values were 0.02, 0.21 and 5.42 µg kg-1for RAC, CLB and SAL, respectively, in bovine and swine muscle samples; CCβ values were 0.03, 0.22 and 5.8 µg kg-1for RAC, CLB and SAL, respectively, in bovine and swine muscle samples. Average recoveries fortified with 0.05-7.5 µg kg-1of the studied β-agonist leads around 95%. The method was demonstrated to be suitable for the determination of RAC, CLB and SLB in swine and cattle muscle samples.
authors.affiliation: a Microbioticos Laboratories , Campinas , SP , Brazil.
authors.forename: Alfredo M
authors.initials: AM
authors.lastname: Montes Nino
authors.affiliation: a Microbioticos Laboratories , Campinas , SP , Brazil.
authors.forename: Rodrigo H M M
authors.identifier: http://orcid.org/0000-0003-1873-7564
authors.initials: RHMM
authors.lastname: Granja
authors.affiliation: a Microbioticos Laboratories , Campinas , SP , Brazil.
authors.forename: Karine V G
authors.initials: KVG
authors.lastname: Reche
authors.affiliation: a Microbioticos Laboratories , Campinas , SP , Brazil.
authors.forename: Fabio M
authors.initials: FM
authors.lastname: Giannotti
authors.affiliation: a Microbioticos Laboratories , Campinas , SP , Brazil.
authors.forename: João K G
authors.initials: JKG
authors.lastname: de Souza
authors.affiliation: b Department of Support Production and Animal Health, School of Medicine Veterinary , São Paulo State University - UNESP , Araçatuba , SP , Brazil.
authors.forename: Sarita P G
authors.initials: SPG
authors.lastname: Ferrari
authors.affiliation: b Department of Support Production and Animal Health, School of Medicine Veterinary , São Paulo State University - UNESP , Araçatuba , SP , Brazil.
authors.forename: André D
authors.initials: AD
authors.lastname: Dos Santos
authors.affiliation: c Department of Structural and Functional Biology, Institute of Biology , University of Campinas - UNICAMP , Campinas , SP , Brazil.
authors.forename: Amarylis C B A
authors.initials: ACBA
authors.lastname: Wanschel
authors.affiliation: c Department of Structural and Functional Biology, Institute of Biology , University of Campinas - UNICAMP , Campinas , SP , Brazil.
authors.forename: Alessandro G
authors.initials: AG
authors.lastname: Salerno
mesh_terms: D000420:Albuterol / Q000032:analysis*; D000818:Animals; D001938:Brazil; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002976:Clenbuterol / Q000032:analysis*; D007753:Laboratories; D057230:Limit of Detection; D009132:Muscles / Q000737:chemistry*; D010627:Phenethylamines / Q000032:analysis*; D013552:Swine; D053719:Tandem Mass Spectrometry
chemical_list: D010627:Phenethylamines; C060897:ractopamine; D000420:Albuterol; D002976:Clenbuterol
keywords: LC-MS/MS; Ractopamine; bovine muscle; clenbuterol; salbutamol; swine muscle