Microbioticos Laboratories

Campinas, Brazil

Microbioticos Laboratories

Campinas, Brazil
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Montes Nino A.M.,Microbioticos Laboratories | Granja R.H.M.M.,Microbioticos Laboratories | Reche K.V.G.,Microbioticos Laboratories | Giannotti F.M.,Microbioticos Laboratories | And 5 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2017

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–1 for RAC, CLB and SAL, respectively. Samples were quantified using RAC-d5, CLB-d9 and SLB-d6 as 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–1 for RAC, CLB and SAL, respectively, in bovine and swine muscle samples; CCβ values were 0.03, 0.22 and 5.8 µg kg–1 for RAC, CLB and SAL, respectively, in bovine and swine muscle samples. Average recoveries fortified with 0.05–7.5 µg kg–1 of 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. © 2017 Informa UK Limited, trading as Taylor & Francis Group.


Granja R.H.M.M.,Microbioticos Laboratories | de Lima A.C.,Microbioticos Laboratories | Patel R.K.,International Atomic Energy Agency | Salerno A.G.,Microbioticos Laboratories | And 2 more authors.
Drug Testing and Analysis | Year: 2012

Florfenicol, a derivative of thiamphenicol in which the hydroxyl group at C-3 has been replaced with fluorine, is listed by the World Health Organization as an antibacterial agent for human medicine that is critically important in risk management strategies for non-human use. AOAC International has also identified it as an important molecule for the development of effective methods for the seafood sector. Following inspection missions from the European Union and United States of America, it was introduced in the Brazilian Residues Control Program to fulfill export and internal national requirements with a Maximum Residue Limit of 800μg/kg. A high performance liquid chromatography method with ultraviolet detection at a wavelength of 230nm (λ=230nm) for the detection of florfenicol in fish muscle was developed and validated according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). Fish samples were extracted with ethyl acetate and hexane followed by C18 solid phase clean-up and chromatographic separation on a reversed-phase C18 LC column with acetonitrile:water as mobile phase. The method results were also was compared with those obtained using liquid chromatography-tandem quadrupole mass spectrometry. The method meets the Brazilian regulatory requirements with a decision limit (CCα) of 840μg/kg and detection capability (CCβ) of 879μg/kg. This method is easy to use and has been implemented into Brazil's residue control program, with liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmation of any suspect samples using the same method. © 2012 John Wiley & Sons, Ltd.


Granja R.H.M.M.,Microbioticos Laboratories | Nino A.M.M.,Microbioticos Laboratories | Reche K.V.G.,Microbioticos Laboratories | Giannotti F.M.,Microbioticos Laboratories | And 4 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2013

Nitroimidazoles are a class of veterinary drugs used for the treatment and prevention of certain bacterial and protozoal diseases in poultry, swine dysentery and genital trichomoniasis in cattle. Since the safety assessment of nitroimidazoles showed them to be genotoxic, carcinogenic and mutagenic, a number of nitroimidazoles have been banned for therapeutic purposes in different countries. Moreover, nitroimidazoles have also been extensively used as growth promoters in food-producing animals. Due to their efficacious improvement in meat production and feed conversion, deliberate use still exists. Therefore, the illegal use of nitroimidazoles in animal husbandry must be monitored. A sensitive method based on LC-MS/MS for the simultaneous determination and confirmation of five banned nitroimidazole drugs including metronidazole, ronidazole, dimetridazole, metronidazole-OH (metabolite of metronidazole), and 2-hydroxymethyl-1-methyl-5-nitroimidazole (metabolite of ronidazole and dimetridazole) in bovine muscle, using ronidazole-d3 as an internal standard, was developed and validated. After extraction with ethyl acetate and evaporation, the nitroimidazoles were reconstituted in petroleum ether and purified, and LC-MS/MS analysis was performed. The method was validated according to Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). Parameters such as decision limit (CCα), detection capability (CCβ), precision, accuracy, uncertaincy and ruggedness were determined. Average accuracy of the five nitroimidazoles from bovine muscle fortified at 5 levels (0.5, 1.0, 1.5, 2.0 and 2.5 μg kg-1) ranged from 96% to 103%. The calculated CCα ranged from 0.0 to 0.17 μg kg-1; CCβ ranged from 0.08 to 0.41 μg kg-1. A complete statistical analysis was performed and the results indicate that the method is robust when subjected to day-to-day analytical variations. © 2013 Copyright Taylor & Francis.


Granja R.H.M.M.,Microbioticos Laboratories | De Lima A.C.,Microbioticos Laboratories | Salerno A.G.,Microbioticos Laboratories | Salerno A.G.,University of Campinas | Wanschel A.C.B.A.,University of Campinas
Journal of AOAC International | Year: 2013

Sulfonamides are one class of antimicrobial agents used in aquaculture production. Sulfonamides are often overused because they are inexpensive and readily available. Their presence at a concentration above the legal limits is a potential hazard to human health. Brazilian authorities have included in the National Regulatory Monitoring Program the control of the three most widely used sulfonamides in aquaculture production, i.e., sulfathiazole, sulfamethazine, and sulfadimethoxine. An LC method with UV detection for the determination of residual sulfonamides in fish muscle, using sulfapyridine as an internal standard has been developed and validated. The validation was performed according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). The method meets the Brazilian regulatory requirement that establishes criteria and procedures for determination of parameters such as decision limit (CCα), detection capability (CCβ), precision, and recovery. For fish muscle, CCα was determined at 3.63, 2.91, and 7.46 μg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. CCβ was 9.39, 14.54, and 9.39 μg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. For shrimp, CCα was 11.5, 8.67, and 4.46 μg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. CCβ was 18, 11.93, and 5.24 μg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. A complete statistical analysis was performed on the results obtained. The results indicate that the method is robust when subjected to day-to-day analytical variations.


Granja R.H.M.M.,Microbioticos Laboratories | de Lima A.C.,Microbioticos Laboratories | Salerno A.G.,Microbioticos Laboratories | Salerno A.G.,University of Campinas | Wanschel A.C.B.A.,University of Campinas
Food Control | Year: 2012

Sulfonamides are one class of antimicrobial agents used in milk production. Sulfonamides are often overused because they are inexpensive and readily available. Their presence at a concentration above the limit value is a potential hazard to human health. Brazilian authorities have included in the National Regulatory Monitoring Program, the control of the sulfonamides in bovine milk production. A simple, rapid, sensitive and reliable high-performance liquid chromatographic method with ultraviolet detection for the simultaneous determination of three sulfonamides (sulfathiazole, sulfamethazine, sulfadimethoxine) in bovine milk, using sulfapyridine as an internal standard has been developed and validated. The whole procedure was validated according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). This Regulation establishes criteria and procedures for determination of parameters such as decision limit (CCα), detection capability (CCβ), precision, recovery and ruggedness. The CCα was determined at 104.4, 102.2 and 105.0 μg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. CCβ was 108.8, 104.9 and 109.9 μg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. Average recoveries fortified with 50, 100 and 200 μg/kg leads around 100%. A complete statistical analysis was performed on the results obtained and the results indicate that the method is robust when subjected to day-to-day analytical variations. © 2012 Elsevier Ltd.


Granja R.H.M.M.,Microbioticos Laboratories | Salerno A.G.,University of Campinas | De Lima A.C.,Microbioticos Laboratories | Montalvo C.,Microbioticos Laboratories | And 3 more authors.
Journal of AOAC International | Year: 2014

Boldenone, an androgenic steroid, is forbidden for use in meat production in most countries worldwide. Residues of this drug in food present a potential risk to consumers. A sensitive LC/MS/MS method for analysis of 17β-boldenone using boldenone-d3 as an internal standard was developed. An enzymatic hydrolysis and extraction using ethyl acetate, methanol, and hexane were performed in the sample preparation. Parameters such as decision limit (CCα), detection capability (CCβ), precision, recovery, and ruggedness were evaluated according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC) and International Organization for Standardization/ International Electrotechnical Commission 17025:2005. CCα and CCβ were determined to be 0.17 and 0.29 μg/kg, respectively. Average recoveries from bovine liver samples fortified with 1, 1.5, and 2 μg/kg were around 100%. A complete statistical analysis was performed on the results obtained, including an estimation of the method uncertainty. The method is considered robust after being subjected to day-to-day analytical variations and has been used as a standard method in Brazil to report boldenone levels in bovine liver.


Sulfonamides are one class of antimicrobial agents used in aquaculture production. Sulfonamides are often overused because they are inexpensive and readily available. Their presence at a concentration above the legal limits is a potential hazard to human health. Brazilian authorities have included in the National Regulatory Monitoring Program the control of the three most widely used sulfonamides in aquaculture production, i.e., sulfathiazole, sulfamethazine, and sulfadimethoxine. An LC method with UV detection for the determination of residual sulfonamides in fish muscle, using sulfapyridine as an internal standard has been developed and validated. The validation was performed according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC). The method meets the Brazilian regulatory requirement that establishes criteria and procedures for determination of parameters such as decision limit (CCalpha), detection capability (CCbeta), precision, and recovery. For fish muscle, CCalpha was determined at 3.63, 2.91, and 7.46 microg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. CCbeta was 9.39, 14.54, and 9.39 microg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. For shrimp, CCalpha was 11.5, 8.67, and 4.46 microg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. CCbeta was 18, 11.93, and 5.24 microg/kg for sulfathiazole, sulfamethazine, and sulfadimethoxine, respectively. A complete statistical analysis was performed on the results obtained. The results indicate that the method is robust when subjected to day-to-day analytical variations.


PubMed | Microbioticos Laboratories
Type: Journal Article | Journal: Journal of AOAC International | Year: 2015

Boldenone, an androgenic steroid, is forbidden for use in meat production in most countries worldwide. Residues of this drug in food present a potential risk to consumers. A sensitive LC/MS/MS method for analysis of 17-boldenone using boldenone-d3 as an internal standard was developed. An enzymatic hydrolysis and extraction using ethyl acetate, methanol, and hexane were performed in the sample preparation. Parameters such as decision limit (CC), detection capability (CC), precision, recovery, and ruggedness were evaluated according to the Brazilian Regulation 24/2009 (equivalent to European Union Decision 2002/657/EC) and International Organization for Standardization/International Electrotechnical Commission 17025:2005. CC and CC were determined to be 0.17 and 0.29 g/kg, respectively. Average recoveries from bovine liver samples fortified with 1, 1.5, and 2 g/kg were around 100%. A complete statistical analysis was performed on the results obtained, including an estimation of the method uncertainty. The method is considered robust after being subjected to day-to-day analytical variations and has been used as a standard method in Brazil to report boldenone levels in bovine liver.

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