Application of the CEN/ISO Standard for Phytase Activity Measurements to a New Phytase Product: Determination of a Robust Conversion Factor by an Interlaboratory Study for Screening Feed Samples

2019 ◽  
Vol 102 (6) ◽  
pp. 1808-1813
Author(s):  
María José González de la Huebra ◽  
Piotr Robouch ◽  
Håkan Emteborg ◽  
Stefano Bellorini ◽  
Aneta Cizek-Stroh ◽  
...  
Keyword(s):  
Conversion Factor ◽  
Feed Additives ◽  
Phytase Activity ◽  
Feed Additive ◽  
The European Union ◽  
Iso Standard ◽  
Official Control ◽  
Activity Measurements ◽  
Feed Samples

Background: Phytase-based preparations are important feed additives currently authorised in the European Union (EU). The European Standard (EN) and International Organization for Standardization (ISO) standard 30024 describes a harmonized method for the determination of phytase activity and is fit-for-purpose for official control of a group of phytase products. However, it is not suitable for the determination of the phytase activity of a new feed additive encoded as 4a16 in the EU Register of Feed Additives, to which a slightly different phytase activity definition has been attributed. Objective: To establish a robust conversion factor to support official control laboratories that apply the EN ISO method when monitoring feed products containing 4a16. Methods: The phytase activity of test materials was determined by the participants using the EN ISO and/or the “applicant” methods. Results: Robust relative SDs for repeatability and for reproducibility of the methods applied for the determination of the phytase activity in the materials containing the 4a16 feed additive ranged from 2.6 to 22% (EN ISO method) and from 2.4 to 39% (applicant method). Conclusions: The data obtained confirmed the performance characteristics published for other phytase-based feeds in the related standard methods. These results allowed us to estimate a factor of 2.68 to convert phytase activities measured with the EN ISO method into the enzyme activity measured with the applicant method. Highlights: The obtained conversion factor will allow EU official laboratories to screen feed samples supplemented with the 4a16 phytase by applying EN ISO Standard 30024.

Application of the CEN/ISO Standard for Phytase Activity Measurements to a New Phytase Product: Determination of a Robust Conversion Factor by an Interlaboratory Study for Screening Feed Samples

2019 ◽  
Vol 102 (6) ◽  
pp. 1808-1813
Author(s):  
María José González de la Huebra ◽  
Piotr Robouch ◽  
Håkan Emteborg ◽  
Stefano Bellorini ◽  
Aneta Cizek-Stroh ◽  
...  
Keyword(s):  
Conversion Factor ◽  
Feed Additives ◽  
Phytase Activity ◽  
Feed Additive ◽  
The European Union ◽  
Iso Standard ◽  
Official Control ◽  
Activity Measurements ◽  
Feed Samples

Abstract Background: Phytase-based preparations are important feed additives currently authorised in the European Union (EU). The European Standard (EN) and International Organization for Standardization (ISO) standard 30024 describes a harmonized method for the determination of phytase activity and is fit-for-purpose for official control of a group of phytase products. However, it is not suitable for the determination of the phytase activity of a new feed additive encoded as 4a16 in the EU Register of Feed Additives, to which a slightly different phytase activity definition has been attributed. Objective: To establish a robust conversion factor to support official control laboratories that apply the EN ISO method when monitoring feed products containing 4a16. Methods: The phytase activity of test materials was determined by the participants using the EN ISO and/or the “applicant” methods. Results: Robust relative SDs for repeatability and for reproducibility of the methods applied for the determination of the phytase activity in the materials containing the 4a16 feed additive ranged from 2.6 to 22% (EN ISO method) and from 2.4 to 39% (applicant method). Conclusions: The data obtained confirmed the performance characteristics published for other phytase-based feeds in the related standard methods. These results allowed us to estimate a factor of 2.68 to convert phytase activities measured with the EN ISO method into the enzyme activity measured with the applicant method. Highlights: The obtained conversion factor will allow EU official laboratories to screen feed samples supplemented with the 4a16 phytase by applying EN ISO Standard 30024.

scholarly journals APPLICATION OF CAPILLARY ELECTROPHORESIS METHOD FOR DETERMINATION OF L-ARGININE CONTENT IN FEED ADDITIVE

2020 ◽  
Vol 21 (1) ◽  
pp. 153-161
Author(s):  
Н. P. Ryvak ◽  
T. R. Levytskyy ◽  
R. O. Ryvak ◽  
S. V. Chorniy
Keyword(s):  
Capillary Electrophoresis ◽  
Test Procedure ◽  
Amino Acid Content ◽  
Feed Additives ◽  
Feed Additive ◽  
Free Form ◽  
Limit Of Quantification ◽  
Test Technique ◽  
Definition Of

The article presents a literature review on the need to balance feed on L-arginine content, its characteristics, ways to supplement the diet of animals and poultry, as well as modern methods for quantitative determination of L-arginine in food, pharmaceuticals, etc. In the section «Materials and methods» the characteristic of the developed test technique, parameters of carrying out research, calibration characteristics with application of a standard sample of L-arginine and carrying out tests of amino acid content in a feed additive by means of system of capillary electrophoresis «Kapel-105M» are given. A description of the validation characteristics performed in the process of validation of the method is given. As a result of the conducted researches the technique of definition of the content of L-arginine in feed additives by means of a method of capillary electrophoresis is developed. The test procedure is based on the dissolution of the feed additive sample, further separation and quantification of the free form of L-arginine, its identification by individual absorption at a wavelength of 200 nm, temperature in the working capillary 30 ºC, voltage 25 kV and conductive electrolyte. The results of validation of this technique by the following characteristics are presented: repeatability, reproducibility, trueness, linearity, limit of quantification, budget uncertainty. The values of trueness, repeatability, reproducibility and uncertainty of the method (with a confidence level of (P) 0.95) does not exceed 5.0 %, the hypothesis of Linearity is acceptable, calculated on the basis of standard deviation (SD) limit of quantification for determining the content of arginine are satisfactory. The results allow us to conclude that the method of capillary electrophoresis using the device «Kapel-105M» is quite accurate and reliable in the case of studies of feed additives L-arginine with a content of the main substance of at least 98,0 %.

scholarly journals Determination of Florfenicol, Thiamfenicol and Chloramfenicol at Trace Levels in Animal Feed by HPLC–MS/MS

Antibiotics ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 59 ◽  
Author(s):  
Rosa Elvira Gavilán ◽  
Carolina Nebot ◽  
Ewelina Patyra ◽  
Beatriz Vazquez ◽  
Jose Manuel Miranda ◽  
...  
Keyword(s):  
Ethyl Acetate ◽  
Animal Feed ◽  
Simultaneous Detection ◽  
The European Union ◽  
European Regulation ◽  
Official Control ◽  
Liquid Chromatography Tandem Mass ◽  
Residue Levels ◽  
Low Levels

Administration of florfenicol and thiamfenicol through medicated feed is permitted within the European Union, always following veterinary prescription and respecting the withdrawal periods. However, the presence of low levels of florfenicol, thiamfenicol, and chloramfenicol in non-target feed is prohibited. Since cross-contamination can occur during the production of medicated feed and according to Annex II of the European Regulation 2019/4/EC, the control of residue levels of florfenicol and thiamfenicol in non-target feed should be monitored and avoided. Based on all the above, a sensitive and reliable method using liquid chromatography tandem mass spectrometry was developed for the simultaneous detection of chloramfenicol, florfenicol, and thiamfenicol at trace levels in animal feed. Analytes were extracted from minced feed with ethyl acetate. Then, the ethyl acetate was evaporated, the residue was resuspended in Milli-Q water and the extract filtered. The method was in-house validated at carryover levels, with concentration ranging from 100 to 1000 µg/kg. The validation was conducted following the European Commission Decision 2002/657/EC and all performance characteristics were successfully satisfied. The capability of the method to detect amfenicols at lower levels than any prior perspective regulation literature guarantees its applicability in official control activities. The developed method has been applied to non-compliant feed samples with satisfactory results.

Interlaboratory Comparison of an Analytical Method for the Determination of Semduramicin in Poultry Feed at the Authorized Level Using High-Performance Liquid Chromatography Coupled to Postcolumn Derivatization and Spectrophotometric Detection

2012 ◽  
Vol 95 (1) ◽  
pp. 61-66 ◽  
Author(s):  
María José González de la Huebra ◽  
Ursula Vincent ◽  
Federica Serano ◽  
Christoph von Holst
Keyword(s):  
Collaborative Study ◽  
Statistical Treatment ◽  
Poultry Feed ◽  
Feed Additive ◽  
Test Material ◽  
The European Union ◽  
Spectrophotometric Detection ◽  
Precision Data ◽  
Official Control ◽  
Postcolumn Derivatization

Abstract The performance characteristics of a method based on HPLC with postcolumn derivatization and spectrophotometric detection for the quantification of semduramicin in poultry feedingstuffs have been determined via a collaborative study. Semduramicin is a feed additive that is authorized for fattening chickens within the European Union at a minimum and maximum content of 20 and 25 mg/kg in feedingstuffs, respectively. The target concentration of semduramicin in the test samples ranged from 11.5 to 45.0 mg/kg. The study has been conducted with two different types of test material, namely, feedingstuff samples that have been previously ground in our laboratory and pelleted feedingstuffs. In the latter case, the laboratories participating in the study had to grind the samples prior to analysis. The obtained RSD for repeatability (RSDr) ranged from 2 to 10% for the ground materials, and from 2 and 7% for the pelleted materials. The RSD for reproducibility (RSDR) varied between 11 and 16% for the ground materials, and between 12 and 15% for the pelleted materials. These data indicated that grinding as an additional step in the analytical procedure did not influence the precision profile of the method. In addition, the HorRat values for all test materials were below or equal to 1.5, thus demonstrating that the obtained precision data were acceptable for the purpose of the method. Furthermore, an estimation of trueness based on statistical treatment of the results reported from the laboratories for spiked samples revealed acceptable mean recovery values of 88 ± 4%. Based on the obtained performance profile, the method can be considered fully validated and transferable to control laboratories to be used within the framework of official control.

scholarly journals Influence of protein-mineral feed additive from marine aquatic organisms on growth intensity and nonspecific resistance of broiler chickens under different microclimate conditions

2020 ◽  
Vol 22 (99) ◽  
pp. 155-160
Author(s):  
N. I. Dankevych ◽  
V. Z. Salata
Keyword(s):  
Broiler Chickens ◽  
Bacterial Contamination ◽  
Specific Resistance ◽  
Aquatic Organisms ◽  
Feed Additives ◽  
Feed Additive ◽  
Growth Intensity ◽  
Microclimate Conditions ◽  
Broiler House

The conducted research was aimed at determination of the impact produced by feeding protein and mineral feed additive produced out of the primary processing aquatic organisms wastes: sea mussels, red algae as well as of the sea water upon the productivity and non-specific resistance of broiler chickens raised in conditions of the normative and non-normative characteristics of the broiler house microclimate. The feed additive was applied to 20–42 day broiler chickens of “Ross 308” cross. The studied broilers were clinically healthy. Throughout the entire experiment, a series of the sanitary and hygienic microclimate parameters were determined, such as temperature, humidity, rate of changes as well as bacterial contamination of air, content of ammonia and carbon dioxide in air, and illumination of the broiler house. The house temperature was measured every day with the aid of a common spirit-in glass thermometer. Air humidity was established with the aid of an August psychrometer, air draft speed, harmful gas concentrations and illumination indicators were measured in compliance with the generally accepted methods. Bacterial contamination was determined with the use of the method of microorganism precipitation on a solid breeding ground placed in Petri dishes followed by a count of the bacterial colonies per 1 m3. The blood analysis included determination of haemoglobin, erythrocytes and leukocytes. In the blood serum, the lysozyme activity (LABS) and bactericidal activity (BABS) were determined. It was established that enriching the basic ration with the protein and mineral additive in quantity of 7 % in addition to the feed mass under conditions of the normative microclimate produced a positive effect on the growth intensity, livability and non-specific resistance indices of the broiler chickens. Thus, the live weight of broilers was reliably greater by 4.7 % and the livability equalled 100 %. The haemoglobin content was reliably greater by 7.6 %, erythrocytes – by 11.5 %, BABS – by 34.5 % and LABS – by 35.9 % as compared with the control group of broiler chickens. At the same time, when the studied feed additive was fed to broiler chickens kept in the microclimate conditions that did not meet the normative requirements, the reliable difference to the control indices was not established. Hence, the research results have proved that application of the protein and mineral feed additive is effective under the optimal microclimate conditions. High figures of livability and growth intensity of broiler chickens are based on a high resistance which is being formed provided the optimal microclimate and application of feed additives have been provided.

scholarly journals The effect of probiotic additives and Bacillus licheniformis inclusion in the diet on broiler growth

2021 ◽  
Vol 77 (05) ◽  
pp. 6534-2021
Author(s):  
KAROLINA FABIA ◽  
DARIUSZ WOLSKI ◽  
DAMIAN KROPISZ ◽  
RADOSŁAW P. RADZKI ◽  
MAREK BIEŃKO ◽  
...  
Keyword(s):  
Bacillus Licheniformis ◽  
Broiler Chickens ◽  
Pectoral Muscle ◽  
Feed Additives ◽  
The Body ◽  
Probiotic Bacteria ◽  
Feed Additive ◽  
The European Union ◽  
Feed Conversion ◽  
The Mean

The withdrawal of antibiotic growth stimulators as of 1 January 2006 in the European Union countries has forced the search for alternative solutions to improve the health and productivity of poultry. The poultry industry also faces the challenge of developing alternative feeding systems with the restriction or exclusion of coccidiostats. One of them is the use of probiotic strains as feed additives. This study aimed to determine the effect of the use in compound feed of a probiotic containing Bacillus licheniformis on rearing rates and postmortem performance of broiler chickens. The experiment was performed on 8012 unsexed broiler chickens of the Ross 308 line divided into four equal groups. The experimental factors were the addition of a probiotic containing Bacillus licheniformis bacteria and the addition of a coccidiostat. The introduction of probiotic bacteria into compound feed did not increase the body weight of birds, in particular, control weights, and at the end of rearing, but it decreased the feed conversion rate (FCR) and mortality. Addition of probiotic in compound feed did not have a significant effect on bird muscle. The mean weight of pectoral muscle between control (C+, C–) and study (C–BL, C+C–BL) groups did not show statistically significant differences; however, the highest mean weight of the evaluated parameter was visible in the C+ group (0. 665) and the lowest in the C–BL group (0. 623). Similarly to the mean weight of the carcass, also in the mean weight of thigh muscles, the lowest statistically significant values were observed in chickens belonging to the C+C–BL group (vs. C–; P <0.05). Based on the obtained results, it can be acknowledged that the probiotic bacteria Bacillus licheniformis in broiler feed can be a good feed additive to replace antibiotics/coccidiostats. This probiotic has a positive effect on the overall health of birds, contributes to better use of nutrients and stimulates growth and development of broiler chickens.

scholarly journals Simultaneous Determination of Ergot Alkaloids in Swine and Dairy Feeds Using Ultra High-Performance Liquid Chromatography-Tandem Mass Spectrometry

Toxins ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 724
Author(s):  
Saranya Poapolathep ◽  
Narumol Klangkaew ◽  
Zhaowei Zhang ◽  
Mario Giorgi ◽  
Antonio Francesco Logrieco ◽  
...  
Keyword(s):  
High Performance ◽  
Ergot Alkaloids ◽  
The European Union ◽  
Chromatography Tandem Mass Spectrometry ◽  
Occurrence Data ◽  
Performance Recovery ◽  
Liquid Chromatography Tandem Mass ◽  
Swine Feed ◽  
Feed Samples

Ergot alkaloids (EAs) are mycotoxins mainly produced by the fungus Claviceps purpurea. EAs are known to affect the nervous system and to be vasoconstrictors in humans and animals. This work presents recent advances in swine and dairy feeds regarding 11 major EAs, namely ergometrine, ergosine, ergotamine, ergocornine, ergocryptine, ergocristine, ergosinine, ergotaminine, ergocorninine, ergocryptinine, and ergocristinine. A reliable, sensitive, and accurate multiple mycotoxin method, based on extraction with a Mycosep 150 multifunctional column prior to analysis using UHPLC-MS/MS, was validated using samples of swine feed (100) and dairy feed (100) for the 11 targeted EAs. Based on the obtained validation results, this method showed good performance recovery and inter-day and intra-day precision that are in accordance with standard criteria to ensure reliable occurrence data on EA contaminants. More than 49% of the swine feed samples were contaminated with EAs, especially ergocryptine(-ine) (40%) and ergosine (-ine) and ergotamine (-ine) (37%). However, many of the 11 EAs were not detectable in any swine feed samples. In addition, there were contaminated (positive) dairy feed samples, especially for ergocryptine (-ine) (50%), ergosine (-ine) (48%), ergotamine (-ine), and ergocristine (-ine) (49%). The mycotoxin levels in the feed samples in this study almost complied with the European Union regulations.

scholarly journals Titrimetric method determination of the content of the mass part of siliciumin feed additives

2020 ◽  
Vol 10 (1) ◽  
pp. 62-65
Author(s):  
O. V. Moravska ◽  
T. R. Levytskyy ◽  
S. O. Vovk
Keyword(s):  
Mass Fraction ◽  
Feed Additives ◽  
Hot Water ◽  
Feed Additive ◽  
Accuracy Control ◽  
Titrimetric Method ◽  
Mass Part ◽  
Powder Samples ◽  
Quadratic Deviation

The results of the development of the method for determining the mass fraction of silicium in powder samples of feed additives by the titrimetric method are presented in the article. The implementation and validation of the method was carried out with the use of feed additive of mixed type "Mikasil" produced by LLC «Globus», Ukraine. The procedure was reproduced ten times with the determination of the mass fraction of siliciumin two parallel samples. The method is based on the precipitation of silicic acid in the form of silicium-fluoride of potassium, followed by hydrolysis with hot water in the presence of chlorous calcium. The isolated hydro-chloric acid in an amount equivalent to the content of hydro-fluoric acid is titrating with alkali solution in the presence of an indicator. The results of the studies showed that the specified method for determining the mass fraction of silicium meets the standards of accuracy control for this method, namely the standard quadratic deviation (σ) is 0.52, when norm up to 0.7, when determined in two parallel samples. The results obtained indicate that the titrimetric method for determining the mass fraction of silicium in powder samples of feed additives is accurate, reliable, reproducible and economically available.

scholarly journals Determination of Phytase Activity in Feed: Interlaboratory Study

2008 ◽  
Vol 91 (2) ◽  
pp. 259-267 ◽  
Author(s):  
Gisele Gizzi ◽  
Peter Thyregod ◽  
Christoph von Holst ◽  
Gerard Bertin ◽  
Kurt Vogel ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Inorganic Phosphate ◽  
Phytase Activity ◽  
Experimental Designs ◽  
Interlaboratory Study ◽  
Intermediate Precision ◽  
Relative Standard ◽  
Feed Samples ◽  
Assay Conditions

Abstract An interlaboratory study was conducted to determine the performance characteristics of a new method for the determination of phytase activity in feed samples. The method is based on the principle that inorganic phosphate is released from the substrate phytate under defined assay conditions and has been validated for its suitability to measure the enzyme activity of various phytase products. Two different experimental designs of the study were applied, allowing for the estimation of the precision of the method under repeatability, intermediate precision and reproducibility conditions, respectively. The relative standard deviation for repeatability (RSDr) ranged from 2.2 to 10.6 and the RSD for reproducibility (RSDR) ranged from 5.4 to 15. The suitability of the validated method for the intended purpose was demonstrated. The obtained performance profile of the method validated in this study was comparable to that of similar methods that were exclusively validated for one phytase product.

Colorimetric Determination of Ronidazole in Feeds

1969 ◽  
Vol 52 (1) ◽  
pp. 101-107
Author(s):  
C R Szalkowski ◽  
J Kanora
Keyword(s):  
Standard Deviation ◽  
Feed Additives ◽  
Colorimetric Determination ◽  
Aminobenzoic Acid ◽  
Colored Complex ◽  
Poultry Feeds ◽  
Naoh Solution ◽  
Significant Interference ◽  
Feed Samples

Abstract Ronidazole, (l-melhyl-5-nitroimidazoI- 2-yl)mcthyl carbamate, is extracted from finished poultry feeds with methanol and chromatographcd on an AL2O3-FIorex column. The eluate is concentrated and dissolved in CCI4 and the ronidazole is extracted with. H2O. The purified ronidazole is hydrolyzcd in NaOH solution in the presence of copper and p-aminobenzoic acid; the resulting diazonium compound is then coupled with naphthylethylenediamine to form a colored complex, which is measured spectrophotometrically at 550 mµ. No significant interference was found from finished feeds, domestic and foreign, or from 48 common feed additives. Recovery experiments on 36 laboratory-prepared feed samples at levels ranging from 0.0015 to 0.030% ronidazole averaged 100.8% with a standard deviation of 2.4.

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