Determination of Fructan (Inulin, FOS, Levan, and Branched Fructan) in Animal Food (Animal Feed, Pet Food, and Ingredients): Single-Laboratory Validation, First Action 2018.07

2019 ◽  
Vol 102 (3) ◽  
pp. 883-892 ◽  
Author(s):  
Barry V McCleary ◽  
Lucie M J Charmier ◽  
Vincent A McKie ◽  
Ciara McLoughlin ◽  
Artur Rogowski
Keyword(s):  
Animal Feed ◽  
Intermediate Precision ◽  
Breeding Parameters ◽  
Animal Feeds ◽  
Test Kit ◽  
Color Response ◽  
Hydrolysis Of ◽  
Single Laboratory ◽  
Agave Fructan

Abstract Traditional enzyme-based methods for measurement of fructan were designed to measure just inulin and branched-type (agave) fructans. The enzymes employed, namely exo-inulinase and endo-inulinase, give incompletely hydrolysis of levan. Levan hydrolysis requires a third enzyme, endo-levanase. This paper describes a method and commercial test kit (Megazyme Fructan Assay Kit) for the determination of all types of fructan (inulin, levan, and branched) in a variety of animal feeds and pet foods. The method has been validated in a single laboratory for analysis of pure inulin, agave fructan, levan, and a range of fructan containing samples. Quantification is based on complete hydrolysis of fructan to fructose and glucose by a mixture of exo-inulinase, endo-inulinase, and endo-levanase, followed by measurement of these sugars using the PAHBAH reducing sugar method which gives the same color response with fructose and glucose. Before hydrolysis of fructan, interfering sucrose and starch in the sample are specifically hydrolyzed and removed by borohydride reduction. The single-laboratory validation (SLV) outlined in this document was performed on commercially available inulin (Raftiline) and agave fructan (Frutafit®), levan purified from Timothy grass, two grass samples, a sample of legume hay, two animal feeds and two barley flours, one of which (Barley MAX®) was genetically enriched in fructan through plant breeding. Parameters examined during the validation included working range, target selectivity, recovery, LOD, LOQ, trueness (bias), precision (repeatability and intermediate precision), robustness, and stability. The method is robust, quick, and simple.

Determination of Sodium Fluoroacetate in Dairy Powders by Liquid Chromatography–Tandem Mass Spectrometry: Single-Laboratory Validation, First Action 2015.02

2016 ◽  
Vol 99 (1) ◽  
pp. 242-251 ◽  
Author(s):  
Lauren M Fleury ◽  
Bryan G Scahill ◽  
Rilka Taskova
Keyword(s):  
Infant Formula ◽  
Intermediate Precision ◽  
Method Performance ◽  
Expert Review ◽  
Liquid Chromatography Tandem Mass ◽  
Expert Review Panel ◽  
Single Laboratory ◽  
Lc Tandem Ms ◽  
Dairy Powders

Abstract A single-laboratory validation (SLV) study was conducted for the determination of sodium fluoroacetate in dairy powders by LC-tandem MS (LC-MS/MS). Linearity of response was confirmed by analysis of samples fortified over the concentration range 0.10–100 μg/kg. The LOD was estimated to be 0.028 μg/kg (0.028 ppb) from the SD of the measured concentrations of infant formula samples fortified at 0.10 μg/kg. The corresponding LOQ calculates at 0.085 μg/kg (0.085 ppb), which ensures excellent reliability of quantification at the limit of reporting of 1.0 μg/kg (1 ppb). Repeatability and intermediate precision were estimated from the SD of the recovery of samples fortified at 0.075, 0.10, 0.20, 0.50, 1.0, and 10.0 μg/kg. The previously mentioned method performance values were established using a representative stage 2 (6–12 months) bovine infant formula, and the robustness of the method was tested by the analysis of 107 unique dairy powders and formulations fortified at 1.0 μg/kg. The data collected in this study satisfy the requirements of SLV studies established by the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN), and the method was awarded First Action Official MethodSM status by the AOAC Expert Review Panel on SPIFAN Nutrient Methods (Contaminants) on March 17, 2015.

Determination of Vitamin K1 in Infant, Pediatric, and Adult Nutritionals by HPLC with Fluorescence Detection: Single-Laboratory Validation, First Action 2015.09

2015 ◽  
Vol 98 (5) ◽  
pp. 1382-1389 ◽  
Author(s):  
Mary Bidlack ◽  
Linda D Butler Thompson ◽  
Wesley A Jacobs ◽  
Karen J Schimpf
Keyword(s):  
Fluorescence Detection ◽  
Hplc Method ◽  
Excitation Wavelength ◽  
Vitamin K1 ◽  
Intermediate Precision ◽  
Method Performance ◽  
Performance Requirements ◽  
Normal Phase Hplc ◽  
Single Laboratory

Abstract This normal-phase HPLC method with postcolumn reduction and fluorescence detection allows for the quantitative determination of trans vitamin K1 in infant, pediatric, and adult nutritionals. Vitamin K1 is extracted from products with iso-octane after precipitation of proteins and release of lipids with methanol. Prepared samples are injected onto a silica HPLC column where cis and trans vitamin K1 are separated with an iso-octane–isopropanol mobile phase. The column eluent is mixed with a dilute ethanolic solution of zinc chloride, sodium acetate, and acetic acid, and vitamin K1 is reduced to a fluorescent derivative in a zinc reactor column. The resulting hydroquinone is then detected by fluorescence at an excitation wavelength of 245 nm and an emission wavelength of 440 nm. During a single-laboratory validation of this method, repeatability and intermediate precision ranged from 0.6 to 3.5% RSD and 1.1 to 6.0% RSD, respectively. Mean overspike recoveries ranged from 91.9 to 106%. The method demonstrated good linearity over a standard range of approximately 2–90 μg/L trans vitamin K1 with r2 averaging 0.99995 and average calibration errors of <1%. LOQ and LOD in ready-to-feed nutritionals were estimated to be 0.03 and 0.09 μg/100 g, respectively. The method met AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals Standard Method Performance Requirements® and was approved as a first action method at the 2015 AOAC Mid-Year Meeting.

scholarly journals Determination of Ethanol Concentration in Kombucha Beverages: Single-Laboratory Validation of an Enzymatic Method, First Action Method 2019.08

2020 ◽  
Author(s):  
Ruth Ivory ◽  
Elaine Delaney ◽  
David Mangan ◽  
Barry V McCleary
Keyword(s):  
Fruit Juice ◽  
Limit Of Detection ◽  
Alcoholic Beverage ◽  
Limit Of Quantification ◽  
Easy Method ◽  
Test Kit ◽  
High Possibility ◽  
Single Laboratory ◽  
Low Alcohol

Abstract Kombucha is a fermented, lightly effervescent sweetened black or green tea drink. It is marketed as a functional beverage based on its proposed health benefits. Kombucha is produced by fermenting tea using a “symbiotic colony of bacteria and yeast” (SCOBY). Kombucha is marketed as a non-alcoholic beverage, however due to the production process employed, there is a high possibility that the Kombucha products will contain low levels of ethanol. Kombucha is sold in a raw and unpasteurized form and, if kept at temperatures above 4 °C, the possibility exists that it will continue to ferment, producing ethanol. This possibility of continued fermentation may lead to an increase in ethanol content from levels below 0.5%ABV at time of production to higher levels at time of consumption. Thus, there is a potential for levels rising to greater than 0.5%ABV, the threshold for certification as a non-alcoholic beverage. It is essential that Kombucha manufacturers have the capacity to accurately and quickly test for ethanol in their products.  The Ethanol Assay Kit is an enzymatic test kit developed by Megazyme for the determination of ethanol in a variety of samples. The kit has been validated in a single laboratory for use with Kombucha fermented drinks, fruit juices, and low-alcohol beer samples. The commercially available Ethanol Assay Kit (Megazyme catalogue no. K-ETOH) contains all components required for the analysis. Quantification is based on the oxidation of ethanol to acetaldehyde by alcohol dehydrogenase and further oxidation of acetaldehyde by acetaldehyde dehydrogenase with conversion of NAD+ to NADH. The single laboratory validation (SLV) outlined in this document was performed on a sample set of eight different commercial Kombucha products purchased in Ireland, a set of five Cerilliant aqueous ethanol solutions, two BCR low-alcohol beer reference materials, two alcohol-free beer samples, and two fruit juice samples against SMPR 2016.001 (1). Parameters examined during the validation included Working range, Selectivity, Limit of Detection (LOD), Limit of Quantification (LOQ), Trueness (bias), Precision (reproducibility and repeatability), Robustness, and Stability. The Ethanol Assay is a robust, quick and easy method for the measurement of ethanol in Kombucha. Our data suggests this method is also reliable for similar matrices, such as low-alcohol beer and fruit juice. The assay meets all requirements set out in in AOAC SMPR 2016.001.

scholarly journals Determination of Tetracyclines in Animal Feeds in the Presence of Other Drugs by Thin-Layer Chromatography and Microbiological Method

1996 ◽  
Vol 79 (2) ◽  
pp. 375-379 ◽  
Author(s):  
Pantelis K Markakis
Keyword(s):  
Thin Layer ◽  
Thin Layer Chromatography ◽  
Animal Feed ◽  
Diffusion Method ◽  
Growth Promoter ◽  
Animal Feeds ◽  
Limit Of Quantitation ◽  
Layer Chromatography ◽  
Method Analysis

Abstract This method was developed to separate, detect, and quantitate oxytetracycline (OTC) or chlortetracycline hydrochloride (CTC HCI) in animal feeds in thepresence of 11 otherdrugs: 3 nitrofurans, 2 macrolideantibiotics,3 sulfonamides, 2 coccidiostatics, and 1 antibacterial growth promoter. OTC or CTC HCI was separated from coexisting drugs and detected by thin-layer chromatography, then quantitated microbiologically by the agar diffusion method. Analysis of 125 experimental animal feed samples fortified at 5 levels (7.5-400 ppm) with OTC or CTC HCI and at 1 level (50 ppm) with the rest of the drugs, respectively, gave a limit of quantitation of 1.25 or 0.625 ppm, a recovery of 90.6 or 92.9%, and a coefficient of variation of 2.9-6.1 or 2.3-4.4%.

Determination of Fructans in Infant, Adult, and Pediatric Nutritional Formulas: Single-Laboratory Validation, First Action 2016.06

2016 ◽  
Vol 99 (6) ◽  
pp. 1576-1588 ◽  
Author(s):  
Philip Haselberger ◽  
Wesley A Jacobs
Keyword(s):  
Quantitative Analysis ◽  
Sample Preparation ◽  
Profile Analysis ◽  
Correction Factors ◽  
Intermediate Precision ◽  
Size Category ◽  
Method Performance ◽  
Performance Requirements ◽  
Single Laboratory

Abstract A method for fructan analysis designed to comply with AOAC Standard Method Performance Requirements (SMPR®) 2014.002 is described. It is closely related to existing methods for fructan analysis, including AOAC 997.08 and 999.03, as well as a method previously published by Cuany et al. This new method achieves LOQ of 0.03% fructan on a ready-to-feed (RTF) basis with mean recoveries ranging from 93 to 108% in the presence of up to 9% sucrose (even at the 0.03% level of fructan). Repeatability ranged from 1.09 to 3.67%. Intermediate precision ranged from 2.46 to 6.79%. Sample preparation for quantitative analysis is simplified compared to some of the existing methodologies. The method incorporates a qualitative profile analysis to determine fructan size category. This allows assignment of appropriate correction factors without independent knowledge of fructan type.

scholarly journals Interlaboratory Comparison of Methods Determining the Botanical Composition of Animal Feed

2018 ◽  
Vol 101 (1) ◽  
pp. 227-234 ◽  
Author(s):  
Luca Braglia ◽  
Laura Morello ◽  
Floriana Gavazzi ◽  
Silvia Gianì ◽  
Francesco Mastromauro ◽  
...  
Keyword(s):  
Interlaboratory Comparison ◽  
Dna Analysis ◽  
Animal Feed ◽  
Optical Microscope ◽  
Botanical Composition ◽  
Animal Feeds ◽  
Interlaboratory Trial ◽  
Quantitative Identification ◽  
Set Up

Abstract A consortium of European enterprises and research institutions has been engaged in the Feed-Code Project with the aim of addressing the requirements stated in European Union Regulation No. 767/2009, concerning market placement and use of feed of known and ascertained botanical composition. Accordingly, an interlaboratory trial was set up to compare the performance of different assays based either on optical microscope or DNA analysis for the qualitative and quantitative identification of the composition of compound animal feeds. A tubulin-based polymorphism method, on which the Feed-Code platform was developed, provided the most accurate results. The present study highlights the need for the performance of ring trials for the determination of the botanical composition of animal feeds and raises an alarm on the actual status of analytical inaccuracy.

Determination of Ochratoxin A in Animal Feed and Cereal Grains by Liquid Chromatography with Fluorescence Detection

1986 ◽  
Vol 69 (6) ◽  
pp. 957-959 ◽  
Author(s):  
Huguette Cohen ◽  
Michel Lapointe
Keyword(s):  
Acetic Acid ◽  
Liquid Chromatography ◽  
Ochratoxin A ◽  
Fluorescence Detection ◽  
Animal Feed ◽  
Cereal Grains ◽  
Fluorescence Detector ◽  
Animal Feeds ◽  
Liquid Chromatographic

Abstract A liquid chromatographic (LC) method is described for determination of ochratoxin A in animal feeds and cereal grains. Samples are initially extracted with chloroform-ethanol (8 + 2) and 5% acetic acid in water. Extracts are purified using a silica gel cartridge followed by a cyano cartridge. The samples are evaporated, diluted to a known volume, and analyzed using a 10 cm column of 3 μm Cm and a fluorescence detector. The method was applied to a variety of animal feeds and cereal grains at levels of 1.0-0.005 ppm added ochratoxin A. The overall recovery was 90.6% ± 3.6.

scholarly journals Determination of Lipophilic Toxins by LC/MS/MS: Single-Laboratory Validation

2011 ◽  
Vol 94 (3) ◽  
pp. 909-922 ◽  
Author(s):  
Adriano Villar-González ◽  
María Luisa Rodríguez-Velasco ◽  
Ana Gagoo-Martínez
Keyword(s):  
Direct Analysis ◽  
Validation Process ◽  
Sample Extraction ◽  
European Union Legislation ◽  
Lipophilic Toxins ◽  
Hydrolysis Of ◽  
Single Laboratory ◽  
Duplicate Sample ◽  
Validation Experiments

Abstract An LC/MS/MS method has been developed, assessed, and intralaboratory-validated for the analysis of the lipophilic toxins currently regulated by European Union legislation: okadaic acid (OA) and dinophysistoxins 1 and 2, including their ester forms; azaspiracids 1, 2, and 3; pectenotoxins 1 and 2; yessotoxin (YTX), and the analogs 45 OH-YTX, Homo YTX, and 45 OH-Homo YTX; as well as for the analysis of 13-desmetil-spirolide C. The method consists of duplicate sample extraction with methanol and direct analysis of the crude extract without further cleanup or concentration. Ester forms of OA and dinophysistoxins are detected as the parent ions after alkaline hydrolysis of the extract. The validation process of this method was performed using both fortified and naturally contaminated samples, and experiments were designed according to International Organization for Standardization, International Union of Pure and Applied Chemistry, and AOAC guidelines. With the exception of YTX in fortified samples, RSDr below 15% and RSDR were below 25%. Recovery values were between 77 and 95%, and LOQs were below 60 μg/kg. These data together with validation experiments for recovery, selectivity, robustness, traceability, and linearity, as well as uncertainty calculations, are presented in this paper.

Determination of Biotin in Infant, Pediatric, and Adult Nutritionals by High-Performance Liquid Chromatography and Fluorescence Detection: Single-Laboratory Validation, First Action 2016.11

2017 ◽  
Vol 100 (1) ◽  
pp. 145-151 ◽  
Author(s):  
Qi Lin ◽  
Yi Ding ◽  
Fiona Poh ◽  
Chunyan Zhang ◽  
Shang-Jing Pan ◽  
...  
Keyword(s):  
Fluorescence Detection ◽  
High Performance ◽  
Reversed Phase ◽  
Hplc Method ◽  
Intermediate Precision ◽  
Metaphosphoric Acid ◽  
Expert Review ◽  
Expert Review Panel ◽  
Single Laboratory

Abstract A reversed-phase HPLC method with postcolumn protein conjugation and fluorescence detection for the quantitative determination of biotin in infant, pediatric, and adult nutritionals was developed and evaluated in a single-laboratory validation (SLV). Sample of appropriate size is mixed with 2% metaphosphoric acid to precipitate out the protein. The filtrate is injected onto a C18 HPLC column in which biotin and riboflavin are separated with an appropriate mobile phase. The biotin, after eluting from the column, binds with the streptavidin fluorescein to become a fluorescent conjugate. The conjugate is then detected by fluorescence at λex = 495 nm and λem = 518 nm. A column switch is used in the method as an option to shorten the run time from 30 to 15 min, by eluting out riboflavin at a higher flow rate. In this SLV, a total of 19 AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals matrixes representing a range of infant,pediatric, and adult formulas were evaluated for their biotin content. The analytical range was 1.66–142 μg/100 g reconstituted final product. The repeatability and intermediate precision ranged from 0.5 to 3.0% RSDr and from 1.3 to 4.5% RSDiR, respectively. Recovery from spiked matrixes varied from 95 to 111%, and accuracy of quantification using Standard Reference Material 1849a ranged from 99 to 105%. The LOQ in reconstituted product was estimated to be 0.8 μg/100 g. The method was approved by the Expert Review Panel as First Action at the 2016 AOAC INTERNATIONAL Mid-Year Meeting.

Method for the Determination of Lycopene in Supplements and Raw Material by Reversed-Phase Liquid Chromatography: Single-Laboratory Validation

2008 ◽  
Vol 91 (6) ◽  
pp. 1284-1297 ◽  
Author(s):  
André Müller ◽  
Bernd Pietsch ◽  
Nicole Faccin ◽  
Joseph Schierle ◽  
Edward H Waysek
Keyword(s):  
Dietary Supplements ◽  
High Performance ◽  
Raw Materials ◽  
Reversed Phase ◽  
Raw Material ◽  
Intermediate Precision ◽  
Relative Standard ◽  
Product Forms ◽  
Single Laboratory

Abstract A single-laboratory validation study was conducted for a liquid chromatographic (LC) method for the determination of total and all-trans-lycopene in a variety of dietary supplements and raw materials. Gelatin-based and other water-dispersible beadlets, or tablets, capsules, and softgels containing such product forms, were digested with protease. Alginate formulations and the respective applications were treated with an alkaline sodium EDTA acetate buffer to release lycopene from the matrix. Lycopene and other carotenoids were extracted from the resulting aqueous suspensions with dichloromethane and ethanol. Oily product forms were directly dissolved in dichloromethane and ethanol. The extracts were chromatographed on an isocratic high-performance LC system using a C16 alkylamide modified silica column that provided satisfactory resolution of all-trans-lycopene from its predominant cis-isomers and separated the lycopene isomers from other carotenoids such as - and -carotene, cryptoxanthin, lutein, and zeaxanthin. The within-day precision relative standard deviation (RSD) for the determination of total lycopene ranged from 0.9 to 5.7 over concentration ranges of 50200 g/kg for raw materials and 0.324 g/kg for dietary supplements. The intermediate precision RSD (total RSD) ranged from 0.8 to 8.9. Recoveries obtained for beadlet and tablet material for the different extraction variants ranged from 95.0 to 102.1 at levels of 0.0220 g/kg for tablets and from 95.0 to 101.1 at levels of 1200 g/kg for beadlet material.

Sign in / Sign up

Export Citation Format

Share Document