Brunelle Biotech Consulting

Woodinville, WA, United States

Brunelle Biotech Consulting

Woodinville, WA, United States
SEARCH FILTERS
Time filter
Source Type

Bapanpally C.,SA Scientific Ltd | Montier L.,SA Scientific Ltd | Khan S.,SA Scientific Ltd | Kasra A.,SA Scientific Ltd | And 4 more authors.
Journal of AOAC International | Year: 2014

The SASTM Molecular tests Escherichia coli O157 Detection method, a loop-mediated isothermal amplification method, performed as well as or better than the U.S. Department of Agriculture, Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, bagged mixed lettuce, and fresh spinach. Ground beef (30% fat, 25 g test portion) was validated for 7-8 h enrichment, leafy greens were validated in a 6-7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16-20 h enrichment. The method performance for meat and leafy green matrixes was also shown to be acceptable under conditions of co-enrichment with Salmonella. Thus, after a short co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. The SAS Molecular tests Salmonella Detection Kit was validated using the same test portions as for the SAS Molecular tests E. coli O157 Detection Kit and those results are presented in a separate report. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 50 E. coli O157 strains, including H7 and non-motile strains, and 30 non-E. coli O157 strains examined. Finally, the method was shown to be robust when variations to DNA extract hold time and DNA volume were varied. The method comparison and robustness data suggest a full 7 h enrichment time should be used for 25 g ground beef test portions.


Bapanpally C.,SA Scientific Ltd | Montier L.,SA Scientific Ltd | Khan S.,SA Scientific Ltd | Kasra A.,SA Scientific Ltd | And 4 more authors.
Journal of AOAC International | Year: 2014

The SASTM Molecular tests Salmonella Detection method, a Loop-mediated Isothermal Amplification method, performed as well as or better than the U.S. Department of Agriculture-Food Safety Inspection Service Microbiology Laboratory Guidebook and the U.S. Food and Drug Administration Bacteriological Analytical Manual reference methods for ground beef, beef trim, ground turkey, chicken carcass rinses, bagged mixed lettuce, and fresh spinach. The ground beef (30% fat, 25 g test portion), poultry matrixes and leafy greens were validated in a 6-7 h enrichment, and ground beef (30% fat, 375 g composite test portion) and beef trim (375 g composite test portion) were validated in a 16-20 h enrichment. The method performance for meat and leafy green matrixes was shown to be acceptable under conditions of co-enrichment with Escherichia coli O157. Thus, after a short 6-7 h co-enrichment step, ground beef, beef trim, lettuce, and spinach can be tested for both Salmonella and E. coli O157. Inclusivity and exclusivity testing revealed no false negatives and no false positives among the 100 Salmonella serovars and 30 non-Salmonella species examined. The method was shown to be robust when enrichment time, DNA extract hold time, and DNA volume were varied.


Burnett T.J.,Elanco Animal Health | Rodewald J.M.,Covance | Moran J.,Elanco Animal Health | Turberg M.P.,Elanco Animal Health | And 2 more authors.
Journal of AOAC International | Year: 2012

A candidate LC method proposed by the Expert Review Panel (ERP) for ractopamine was evaluated in a single-laboratory validation (SLV) study. The matrixes examined included bovine liver, kidney, muscle, and fat; swine liver, kidney, muscle, and fat; and turkey liver and muscle. Solution standards were shown to provide a linear response with an unweighted regression. The method demonstrated acceptable precision (HorRatr values 0.25 to 1.38) and recovery (75.4 to 88.8%) in all fortified matrixes. Method precision was verified with incurred residue tissues (bovine liver, kidney, and muscle; swine liver, kidney, and muscle; and turkey liver and muscle), which yielded RSDr values below 16% for all tissues and below 7% for most tissues. Estimated LOQ values ranged from 1.8 to 20.7 ng/g and support the utility of the method in the range of the maximum residue limits or tolerances for the various tissues. The data satisfy the requirements of the AOAC Stakeholder Panel on Veterinary Drug Residue for SLV studies, and the method was adopted Official Methods of Analysis SM First Action 2011.22 by the AOAC ERP on Veterinary Drug Residues.


Coleman M.R.,Elanco Animal Health | Rodewald J.M.,Covance | Brunelle S.L.,Brunelle Biotech Consulting | Nelson M.,Independent Consultant | And 2 more authors.
Journal of AOAC International | Year: 2014

A single-laboratory validation (SLV) study was conducted on an LC/MS/MS method for the determination and confirmation of nicarbazin, expressed as 4,4-dinitrocarbanilide (DNC), in chicken tissues, including liver, kidney, muscle, skin with adhering fat, and eggs. Linearity was demonstrated with DNC standard curve solutions using a weighted (1/x) regression and confirmed with matrix-matched standards. Intertrial repeatability precision (relative standard deviation of repeatability; RSDr) was from 2.5 to 11.3%, as determined in fortified tissues. The precision was verified with incurred tissue, and varied from 0.53 to 2.5%. Average recoveries ranged from 82% in egg to 98% in kidney. Although the average recoveries across all concentrations were within the acceptable range, the method was improved with the inclusion of an internal standard and the use of matrix-matched standards. Accuracy for the improved method in chicken liver varied from 93 to 99% across all concentrations (100-8000 ng/g) compared to recoveries below 80% at concentrations between 100-400 ng/g in chicken liver for the original method. The limit of detection was estimated to be less than 3.0 ng/g in all tissue types, and the limit of quantitation was validated at 20 ng/g. Based on confirmatory ion ratios and peak retention times, the false-negative rate was estimated as 0.00% (95% confidence limits 0.00, 0.74%) from 484 fortified samples and 12 incurred residue samples analyzed using the U.S. and EU confirmation criteria. Small variations to the method parameters, with the exception of injection volume, did not have a significant effect on recoveries. Stability was determined for fortified tissues, extracts, and standard curve solutions. The data collected in this study satisfy the requirements of SLV studies established by the AOAC Stakeholder Panel for Veterinary Drug Residue and the method was awarded First Action Official MethodSM status by the Expert Review Panel for Veterinary Drug Residues on May 7, 2013. © 2014 Publishing Technology.


Mizinga K.M.,Elanco Animal Health | Burnett T.J.,Elanco Animal Health | Brunelle S.L.,Brunelle Biotech Consulting | Wallace M.A.,MPI Research Inc | Coleman M.R.,Brunelle Biotech Consulting
Journal of AOAC International | Year: 2016

Lilly Method AM-AA-CA-R108-AB-755, which is substantially the same as U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS) Chemistry Laboratory Guidebook (CLG) method R22, is the current regulatory method for determining narasin in cattle and chicken tissues and is based on bioautography, creating a zone of inhibition of bacterial growth, with the size of the zone correlating to the amount of narasin extracted from the tissue. AOAC Method 2011.24 is an LC-Tandem mass spectrometry (MS/MS) method for determining narasin content from bovine, swine, or chicken tissues. It has many advantages over the regulatory method, including higher throughput, less solvent use, no use of carbon tetrachloride, a wider method range, inclusion of swine tissues, and it is less labor intensive. In this study, AOAC Method 2011.24 was compared to FSIS CLG method R22 for the determination of narasin in chicken abdominal fat. Fortified chicken-fat samples ranging from 20 to 960 ng/g and incurred chicken-fat samples ranging from 40 to 480 ng/g were assayed by both methods in triplicate. Mean accuracies for the two methods were similar, 77-110% for CLG R22 and 84-96% for AOAC Method 2011.24, and the method results showed a linear correlation. The methods differed in precision, however, with the CLG R22 method yielding 2.6-34% RSD and AOAC Method 2011.24 yielding 0.15-6.4% RSD. It is recommended that AOAC Method 2011.24-granted AOAC Official MethodSM Final Action status-be adopted as the official U.S. regulatory method.


Burnett T.J.,Elanco Animal Health | Rodewald J.M.,Covance | Brunelle S.L.,Brunelle Biotech Consulting | Neeley M.,MPI Research | And 3 more authors.
Journal of AOAC International | Year: 2012

A candidate method selected by the AOAC Expert Review Panel (ERP) for Ractopamine for determination and confirmation of parent and total ractopamine by LC/MS/MS was validated in a single laboratory for bovine, swine, and turkey tissues. The candidate method utilizes methanol extraction of the tissues, followed by an optional enzymatic hydrolysis for determination of total (parent plus conjugate) ractopamine. A mixed-mode cation exchange SPE cartridge is used to purify the initial extract before LC/MS/MS. Matrix-matched standards and a ractopamine-d6 internal standard are used for quantification of parent and total ractopamine in unknown samples. Validation data demonstrated that mean intertrial recoveries for ractopamine across all concentrations tested ranged from 79.7 to 102.2% for parent ractopamine and from 79.0 to 100.0% when a hydrolysis step was included. Intertrial repeatability precision ranged from 2.44 to 11.1% for parent ractopamine and 4.97 to 15.0% with hydrolysis. Estimated LOD values were below 0.1 ng/g and LOQ values were validated at 0.25x the maximum residue limits. The data satisfy the requirements of the AOAC Stakeholder Panel for Veterinary Drug Residue Methods for single laboratory validation studies. The method was awarded Official Methods of Analysis SM First Action 2011.23 by the AOAC ERP on Veterinary Drug Residues.


Bolong W.,Chinese Academy of Inspection and Quarantine | Fengxia Z.,Chinese Academy of Inspection and Quarantine | Xiaoning M.,Chinese Academy of Inspection and Quarantine | Fengjuan Z.,Chinese Academy of Inspection and Quarantine | Brunelle S.L.,Brunelle Biotech Consulting
Journal of AOAC International | Year: 2016

A potentiometric method for determination of chloride was validated against AOAC Standard Method Performance Requirement (SMPR®) 2014.015. Ten AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN ) matrixes, including National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 1849a, were tested in duplicate on 6 independent days. The repeatability (RSDr) ranged from 0.43 to 1.34%, and the intermediate reproducibility (RSDiR) ranged from 0.80 to 3.04%. All results for NIST SRM 1849a were within the range of the certified concentration (701 ± 17 mg/100 g). Recovery was demonstrated with two overspike levels, 50 and 100%, in the 10 SPIFAN matrixes. Samples were tested in duplicate on 3 different days, and all results were within the SMPR requirement of 95 to 105%. The LOQs of the method for powdered products and ready-to-feed or reconstituted products were 20 mg/100 g and 2.2 mg/100 mL, respectively. A wide analytical range from the LOQ to 99.5% chlorine content can be reached with an appropriate dilution factor, but in practice, the upper analytical value observed in routine matrix testing was approximately 1080 mg/100 g in skim milk powder. This is a rapid, simple, and reliable chlorine-testing method applicable to infant formula, adult nutritionals, and ingredients used in these dairy-based products, such as skim milk powder, desalted whey powder, whey protein powder, and whole milk powder.


A previously validated method for determination of chondroitin sulfate in raw materials and dietary supplements was submitted to the AOAC Expert Review Panel (ERP) for Stakeholder Panel on Dietary Supplements Set 1 Ingredients (Anthocyanins, Chondroitin, and PDE5 Inhibitors) for consideration of First Action Official MethodsSM status. The ERP evaluated the single-laboratory validation results against AOAC Standard Method Performance Requirements 2014.009. With recoveries of 100.8-101.6% in raw materials and 105.4-105.8% in finished products and precision of 0.25-1.8% RSDr within-day and 1.6-4.72% RSDr overall, the ERP adopted the method for First Action Official Methods status and provided recommendations for achieving Final Action status.


Thompson J.J.,Abbott Laboratories | Pacquette L.,Abbott Laboratories | Brunelle S.L.,Brunelle Biotech Consulting
Journal of AOAC International | Year: 2015

A method for determination of 12 minerals and trace elements (Na, Mg, P, K, Ca, Cr, Mn, Fe, Cu, Zn, Se, and Mo) in infant formula and adult/ pediatric nutritional formula was developed and evaluated in a single-laboratory validation. Some additional reproducibility data were obtained from a small interlaboratory study. The method involves microwave digestion of the sample followed by inductively coupled plasma/MS and uses Ge and Te as internal standards. The method is an extension of Official MethodSM 2011.19 and was compared to AOAC Standard Method Performance Requirements (SMPRs®) 2011.009 and 2014.004 developed by the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN). Repeatability precision for the 12 elements in 11 SPIFAN matrixes and National Institute of Standards and Technology Standard Reference Material (SRM) 1849a was >5%, meeting the SMPR criterion for repeatability. Intermediate reproducibility (8 days, two analysts, two instruments) in the 11 SPIFAN matrixes was >5% for nine (Na, Mg, P, K, Mn, Fe, Cu, Zn, Se) of the 12 elements in all 11 matrixes. The mean reproducibility across 6-7 laboratories and seven SPIFAN matrixes ranged from 2.5% for Cu to 7.1% for P. Recovery from spiked matrixes varied from 90.1 to 109%, and accuracy of determination using SRM 1849a ranged from 96.2 to 107.7%, meeting the requirement of 90-110% recovery/accuracy.


A previously validated method for determination of chondroitin sulfate in raw materials and dietary supplements was submitted to the AOAC Expert Review Panel (ERP) for Stakeholder Panel on Dietary Supplements Set 1 Ingredients (Anthocyanins, Chondroitin, and PDE5 Inhibitors) for consideration of First Action Official Methods(SM) status. The ERP evaluated the single-laboratory validation results against AOAC Standard Method Performance Requirements 2014.009. With recoveries of 100.8-101.6% in raw materials and 105.4-105.8% in finished products and precision of 0.25-1.8% RSDr within-day and 1.6-4.72% RSDr overall, the ERP adopted the method for First Action Official Methods status and provided recommendations for achieving Final Action status.

Loading Brunelle Biotech Consulting collaborators
Loading Brunelle Biotech Consulting collaborators