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Evidence Project Final Report

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Project identification

1. Defra Project code FA0121

2. Project title

Determination of a Nitrogen Factor for Chicken Breast

3. Contractororganisation(s)

Royal Society of Chemistry – Analytical Methods Committee.

54. Total Defra project costs £10,080 +VAT(agreed fixed price)

5. Project: start date................. 20/10/2012

EVID4 Evidence Project Final Report (Rev. 06/11) of 19

mailto:[email protected]

end date.................. 15/07/2013


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they be made public. They should be written in a clear and concise manner and represent a full account of the research project which someone not closely associated with the project can follow.Defra recognises that in a small minority of cases there may be information, such as intellectual property or commercially confidential data, used in or generated by the research project, which should not be disclosed. In these cases, such information should be detailed in a separate annex (not to be published) so that the Evidence Project Final Report can be placed in the public domain. Where it is impossible to complete the Final Report without including references to any sensitive or confidential data, the information should be included and section (b) completed. NB: only in exceptional circumstances will Defra expect contractors to give a "No" answer.In all cases, reasons for withholding information must be fully in line with exemptions under the Environmental Information Regulations or the Freedom of Information Act 2000.

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Executive Summary7. The executive summary must not exceed 2 sides in total of A4 and should be understandable to the intelligent

non-scientist. It should cover the main objectives, methods and findings of the research, together with any other significant events and options for new work.

The last study to determine nitrogen factors for chicken (whole chicken, breast, leg etc) was funded by MAFF and published by the Royal Society of Chemistry – Analytical Methods Committee (AMC) in 2000. In the 2000 study, 120 birds (comprising of Ross and Cobb breeds, male and female, light and heavy broilers) were used. The recommended nitrogen factor for skinless chicken breast arising out of this study was instrumental in exposing the misleading labelling of injected chicken breast investigated by the Food Standards Agency and enforcement authorities. The UK methodology (based on the Stubbs and More method) for analysing chicken preparations and the chicken breast N-factor have also been published in the OJ for use during an EC Coordinated Control Programme analysing injected chicken breast in 2005. The European Commission (DG-AGRI) funded a recent project (Project 30-CE-0460798/00-25) with LGC and other European National Reference Laboratories (NRLs) to re-assess the physiological water content of skinless chicken breast and skin-on legs. This information is required to enforce the Poultrymeat Marketing Standards Regulation (EC) 543/2008, which limits the amount of water, poultry and poultry parts can pick up during preparation in poultry plants. Samples of skinless chicken breast and skin-on legs from Ross and Cobb breeds, male and females, light and heavy broilers were collected from each of the seven main poultry producing countries (UK, Germany, France, Italy, Spain, the Netherlands and Poland). The new breeds of Ross (308) and Cobb (500) have shorter rearing times to reach killing weights than the breeds used in the 2000 AMC study. As the birds now used by industry are younger, the nitrogen factor may have also changed (more water and less protein, as the results of Project 30-CE-0460798/00-25 showed). Given that the samples of Ross and Cobb skinless chicken breast had already been collected, and were analysed by NRLs for water and protein (nitrogen) content, it was considered extremely cost effective to do the additional analyses (fat, ash and hydroxyproline) to enable the nitrogen factor for chicken breast to be compared to the factor derived in the 2000 study. Eighty four samples of homogenised chicken breast from the European Commission study were selected from the 144 UK, Polish and Dutch breast samples, as being most appropriate for the UK market, to give an equal distribution of size, breed and country. The samples were analysed for their ash, fat, and hydroxyproline contents, with the moisture and nitrogen results taken from the European Commission study. The results indicated that the average fat content was slightly


lower than in the 2000 study (1.5g/100g instead of 2.0g/100g), as was the hydroxyproline content (0.05g/100g instead of 0.08g/100g). Hydroxyproline is determined to measure the amount of any added hydrolysed collagen used a water retaining agent in tumbled or injected chicken breast.As the fat and ash contents were low, it was decided that the nitrogen factor could be calculated using all 144 breast samples from the UK, Poland and the Netherlands determined in the European Commission study. The results indicated that the main differences in nitrogen content from the 2000 study were with the chicken breasts from the heavy male birds, which were significantly lower than the previous study. Also in this study, chicken breasts from the heavy male birds were also significantly lower in nitrogen than those from lighter males or the light or heavy female birds. As the UK market for chicken breast is derived from both light and heavy birds and it is not customary to separate male and female birds destined for the broiler market after hatching, the nitrogen content most representative of the market is an average across breed, gender and size. In addition, spent hen meat is no longer used for the manufacture of UK chicken products. The average nitrogen content of all broilers is 3.70g/100g with a standard error of the mean of 0.02. Where the raw material of a product is known to be skinless chicken breast, a fat-free nitrogen factor for skinless chicken breast of 3.75 is recommended. This value has been calculated from an average fat content of 1.5g/100g, rounded to the nearest 0.05 and compares to the recommended fat-free factor in the 2000 study for skinless chicken breast of 3.85, which represents a 3% difference.

Project Report to Defra8. As a guide this report should be no longer than 20 sides of A4. This report is to provide Defra with details of

the outputs of the research project for internal purposes; to meet the terms of the contract; and to allow Defra to publish details of the outputs to meet Environmental Information Regulation or Freedom of Information obligations. This short report to Defra does not preclude contractors from also seeking to publish a full, formal scientific report/paper in an appropriate scientific or other journal/publication. Indeed, Defra actively encourages such publications as part of the contract terms. The report to Defra should include: the objectives as set out in the contract; the extent to which the objectives set out in the contract have been met; details of methods used and the results obtained, including statistical analysis (if appropriate); a discussion of the results and their reliability; the main implications of the findings; possible future work; and any action resulting from the research (e.g. IP, Knowledge Exchange).

FINAL REPORT FOR PROJECT FA0121

Mark Woolfe – RSC-AMC Nitrogen Factors Sub-Committee; Joanna R Topping, Stephen L R Ellison, Selvarani Elahi, - LGC, Teddington.

(Any opinions expressed in this report are those of the authors and not the opinions of either Defra or the European Commission)

Determination of a Nitrogen Factor for Chicken Breast

1.0 OBJECTIVE OF PROJECT

The main objective of this project is to revisit the recommended nitrogen factor for skinless chicken breast. This small project commenced in October 2102 and completion of the analyses was delayed by 2 months because of the volume of work at LGC. The statistical analyses of the results were also


Analytical Division, Analytical Methods Committee

Nitrogen Factors Sub-Committee

delayed by 4 months due to the heavy work programme of the statistician. The milestones are given below:

Target date Monthsfrom start date Description of milestone Status

20/10/12 0 Commission project with LGC, and set up sampling plan with statisticians Completed

20/01/13 3 monthsComplete analyses (fat, ash, hydroxyproline), and obtain results from NRLs for moisture and protein

Completed but delayed till March 13

20/02/13 4 month Complete statistical analyses and write final report

Completed but delayed till July 13

20/04/13 2 months Publish research paper agreed by RSC-AMC NFSC, Defra, DG-AGRI

Completed in Dec 13

2.0 INTRODUCTION

Labelling rules(1) require that where ingredients are highlighted in the name of the food, the amount of that ingredient must be declared as a percentage of the final product (QUID- quantitative ingredient declaration). Although the amount of an ingredient is calculated on a recipe basis, enforcement authorities usually check the declaration by analysis of the finished product. The analysis determines the nitrogen (mainly on a fat free basis) of meat ingredient, and converts this to a meat content using a previously determined nitrogen factor. A considerable amount of research has already been undertaken to determine robust nitrogen factors for the common meat and poultry species, and some fish species. This project revisits the nitrogen factor for skinless chicken breast.

Where relevant, a determination of meat content permits the calculation of added water by difference. National regulations(2) require that where a product looks like a cut or joint of meat or poultry e.g. a chicken breast with added water, if there is more than 5% added water, this has to appear in the name of the food. Also if ingredients are used from a different species e.g. hydrolysed proteins from pork or beef to hold in large amounts of added water in the chicken breast, then they also have to be declared in the name of the food.

2.1 The Market for Chicken BreastThe UK market for chicken is worth over £2 billion per year. Skinless chicken breast is the most used portion in the manufacture of chicken products and in catering. The UK cannot produce sufficient chicken breast to meet demand, and significant quantities of chicken breast are imported from third countries mainly Brazil and Thailand to be used for further processing and in catering. In addition, there is a large trade (over £200million/year) in chicken breast preparations (injected and tumbled with water and water retaining agents) for the catering trade produced in the Netherlands, Poland and, to a lesser extent, the UK. The raw material for injected and tumbled chicken preparations was previously Brazilian chicken breast with 1.2% salt, which was imported into the EU with a lower tariff than unsalted chicken breast. However, the customs classification of salted chicken breast was changed in 2006, and the provisions in the EU Poultrymeat Standard Regulation (part of the EU Marketing Regulation 1234/2007(3)) were amended in 2009(4) to include chicken preparations. One of the consequences has been halting the use of Brazilian salted frozen chicken breast as the raw material for tumbled and injected chicken preparations. The raw material for these preparations now is predominantly the more expensive frozen but unsalted chicken breast from Brazil.

2.2 Rationale for this Project

The last study to determine nitrogen factors for chicken (whole chicken, breast, leg etc) was funded by MAFF and published in 20005. In that study, 120 birds (comprising of Ross and Cobb breeds, male and female, light and heavy broilers) were used. The light birds were reared to 35 days, and the heavy


broilers to 49-56 days. The methodology to measure chicken content and added water in chicken products and preparations uses an accurate, agreed nitrogen factor, and was instrumental in exposing misleading labelling of injected chicken breast investigated by the Food Standards Agency6 and enforcement authorities. The methodologies for analysing chicken preparations and the chicken breast N-factor also have been published in the OJ7. These were used in 2005 during an EC Coordinated Control Programme analysing injected chicken breast.

The European Commission (DG-AGRI) funded a project(8) (Project 30-CE-0460798/00-25) with LGC and other European National Reference Laboratories (NRLs) to re-assess the physiological water content of skinless chicken breast and skin-on legs. This was to facilitate enforcement of the Poultrymeat Marketing Standards Regulation (EC) 543/2008(9) which regulates the amount of extraneous water poultry and poultry parts are allowed to pick up during preparation in poultry plants without any declaration on the product label. In this project, forty eight birds (Ross and Cobb breeds, male and females, light and heavy broilers) were collected from each of the seven main poultry producing European countries (UK, Germany, France, Italy, Spain, the Netherlands and Poland). Samples of skinless breast and skin-on legs were prepared and homogenised. The new chicken breeds of Ross (308) and Cobb (500) have shorter rearing times to reach killing weights than the breeds used in the 2000 AMC study i.e. light weight birds are reared only to 30-32 days and heavy birds only to 48-52 days. As the birds now used by industry are younger, it is possible that the nitrogen factor could have changed (more water and less protein). Given changes in rearing breed and rearing practice, an up to date and accurate nitrogen factor for chicken breast is needed to ensure there is no unfair competition to UK processors and manufacturers, and that there is no misleading labelling of chicken breast preparations in the retail and catering sector. As samples of Ross and Cobb skinless chicken breast had already been collected and analysed for water and protein content as part of the EC study, it was considered extremely cost effective to do the necessary additional analyses (fat, ash and hydroxyproline) to enable the nitrogen factor for chicken breast produced today to be compared to the factor derived in the 2000 study.

3.0 SAMPLE PREPARATION

3.1 Collection and Preparation of Samples

Collection of samples in the DG-AGRI project was under supervision by LGC staff, and according to an SOP developed by LGC to ensure as far as possible that the samples were collected from the 7 countries in the same way and without contact with water. Selected birds were taken off the production line after killing and bleeding, but before entering the scald tank. The skin (with feathers attached) around the breast was removed and the whole breast cut away. The breast muscles were removed from the breast bone and trimmed to remove adhering fat or connective tissue and then double bagged and coded (see Fig. 1). The samples were frozen and sent to LGC where they were homogenised according to European Regulation No. 543/2008 Annex VIII. Each homogenised sample was divided into two aliquots and stored frozen.


3.2 Selection of Samples

The project budget was sufficient for the selection and analysis of 84 chicken breast samples from the samples collected for the DG-AGRI project. The samples were chosen on the basis that they should be representative of those found on the UK market. It was decided to use samples from UK birds or those with a similar weight to the British chickens, and which were likely to be exported to the UK market. A stratified sampling plan was devised by LGC statisticians to cover a selection of equal numbers of light (around 1.8kg) and heavy birds (around 2.4kg) from the UK, Poland and the Netherlands. The sampling was planned to balance breed and weight as these were considered the most important factors affecting nitrogen content. Cobb was included as a breed likely to reach the UK even though Ross currently is predominant in UK production. The resulting samples contained equal numbers of each breed, equal numbers at each weight, and retained a reasonably even ratio of male to female birds (see Table 1).

Table 1 Selection of 84 Chicken Breast SamplesCountry No. of

SamplesSamples by Breed Samples by Gender Samples by Size

UK 28 14 Ross + 14 Cobb 16 Males + 12 Females 14 Light + 14 Heavy

Poland 28 14 Ross + 14 Cobb 13 Males + 15 Females 14 Light + 14 Heavy

The Netherlands 28 14 Ross + 14 Cobb 14 Males + 14 Females 14 Light + 14 Heavy

4.0 METHODS OF ANALYSISResults for moisture and nitrogen content of the 84 samples selected in 3.2 above were those obtained during the DG-AGRI project. The samples were distributed equally to eight European NRLs who analysed each sample for moisture and nitrogen. The methods used were those laid down in Commission Regulation (EC) 543/2008, which are the same ISO (BSI) standard methods used routinely by Official Control Laboratories in the UK:

Analytical Methods for Meat and Meat Products: Part 2 1980 (1993) Determination of nitrogen or equivalent method;

Part 3 1970 (1997) Determination of moisture. Each sample was analysed in triplicate and a certified reference material (ERM®-BB501a) included in each batch.

Agreement between the triplicate results was to be less than 0.4g/100g for moisture and less than 0.16g/100g for nitrogen.


Figure 1 Preparation of Skinless Chicken Breast

The 84 breast samples were analysed in duplicate for ash, fat and hydroxyproline by LGC using the following BSI validated methods or their equivalents:

Part 1 1970 (1993) Determination of ash Part 4 1970 (1993) Determination of total fat Part 11 1995 Determination of L-(-)-Hydroxyproline

The detailed requirements for the analyses included:a) A standard reference material (e.g. FAPAS meat sample or LGC Reference Material 7152,

Processed Pork) was to be run in duplicate with each batch of chicken samples. b) Two replicates of the three analyses were required per sample and the total content of moisture,

ash, fat and protein (nitrogen x 6.25) should add up to 100% ± 2%. c) The duplicates were to be randomised within each batch. Analytical agreement was required

between the duplicates - the repeatability limits were 0.5g/100g (fat) and 0.1g/100g (ash and hydroxyproline).

5.0 RESULTS

5.1 Results of Ash, Fat and Hydroxyproline Analyses

A summary of the 84 chemical analyses is given in Annex 1, and the total content of ash, fat, moisture, and protein (nitrogen x 6.25) was found to be between 98.8% and 101.7% i.e. within the tolerance laid down in the protocol. Table 2 gives the means of the fat, ash , hydroxyproline, moisture and nitrogen contents for all 84 analyses, and compares them with the 2000 study(1) i.e. the mean of the same analytes for skinless breast from light and heavy broilers.

Table 2 Mean Ash, Fat, Hydroxyproline, Moisture and Nitrogen Contents for 84 Chicken Breast Samples

Analyte (g/100g) 2013 Study(Mean of 84 samples)

2000 Study (Mean of 120 samples)

Ash 1.24 1.12

Fat 1.51 2.0

Hydroxyproline 0.05 0.08

Moisture 74.7 74.2

Nitrogen 3.70 3.78

Table 3 Fat (%) by breed, size and gender

BreedSize Gender LSMean SEM

Cobb L F 1.41 0.13Ross L F 1.41 0.13Cobb H F 1.34 0.13Ross H F 1.51 0.13Cobb L M 1.48 0.13Ross L M 1.31 0.13Cobb H M 1.80 0.13Ross H M 1.81 0.13

An overview of the proximate data is shown in Annex 2, ANNEX 22. As expected, the proximates tended to be inversely correlated. There were no serious outliers and no individual observations appear extreme, although one sample is clearly high for ash, and one for moisture. Given that


Table 4 Ash (%) by breed, size and gender

Breed Size Gender LSMean SEMCobb L F 1.26 0.016Ross L F 1.29 0.016Cobb H F 1.25 0.016Ross H F 1.22 0.016Cobb L M 1.24 0.016Ross L M 1.27 0.016Cobb H M 1.22 0.016Ross H M 1.20 0.016

replicate analyses of these samples agree well, all results were retained for the subsequent statistical analysis. Box plots of sample means for each analyte by breed, country, gender and size are shown in Annex 2, to respectively. Inspection of the analytical data shows no appreciable differences between breeds for any analyte. Some differences between samples from different countries of origin can be seen, especially on the fat content for UK samples, which appear appreciably more variable. The UK samples were collected from 3 different plants by three different teams, whereas the Dutch samples were collected by the same team within 1 day of each other, and the Polish samples on the same day by the same team. The differences in fat content may be the result of small differences in the trimming and preparation of the samples.

Table 5 Hydroxyproline (g/100g) by breed, size and gender

Breed Size Gender LSMean SEMCobb L F 0.045 0.0020Ross L F 0.046 0.0020Cobb H F 0.043 0.0020Ross H F 0.045 0.0020Cobb L M 0.050 0.0020Ross L M 0.050 0.0021Cobb H M 0.050 0.0020Ross H M 0.051 0.0020

Tables 3, 4, 5, show data for the mean composition for fat, ash and hydroxyproline contents respectively. All values have been calculated for breed, size and gender for comparison with those generated in the 2000 study. The means shown are least square means (LSMean), that is, means calculated from a linear model, corrected for imbalance. The hydroxyproline values showed no anomalous results. Statistically significant differences were found for the effect of gender on hydroxyproline content. As expected, size of the chicken breast had a significant effect on values for ash content. The breed of chicken was not a statistically significant variable at the 95% level of confidence for any analyte.

Examining the overall means from the 84 samples, the data in Table 1 shows that there has been a small reduction in ash, fat, hydroxyproline and nitrogen contents when compared to the results found in the 2000 study. The main contributor to the reduction in nitrogen content is from heavy male birds, where it would be expected that there would be a higher nitrogen content than with younger birds.

As the mean fat and ash contents are very low, and moisture and protein (nitrogen) contents contribute more than 97.8% of the chicken breast by weight, it was decided to increase the sample size from 84 to 144. This was done by utilising the moisture and nitrogen results from all the chicken breast samples from the UK, Poland and the Netherlands (48 from each country) used in the DG-AGRI study. This increased the confidence of the nitrogen content results because there was a 71% increase in sample numbers.

5.2 Results from Analyses of 144 samples for Nitrogen and Moisture

A scatter plot of the means (triplicate determinations) of moisture versus nitrogen content for all 144 samples is shown in Annex 3, Figure 7. There appear to be no serious outlying samples, and all samples were retained in the calculations. Figures 8 to 11 in Annex 3 show box plots of moisture and nitrogen contents by gender, size, breed and country respectively. From these data it can be seen that there are some appreciable differences for the nitrogen content by gender and size.

The data in Tables 6 to 9 show the estimated mean contents for moisture and nitrogen for different categories of sample. As in Section 5.1, the means shown are least square means (LSMean). The


data in Tables 6 and 7 show that gender is strongly significant (p < 0.01) for moisture, and very strongly significant for nitrogen (p < 0.001). In addition, size is very strongly significant (p < 0.001) for nitrogen and significant for moisture. Breed was statistically significant at the 95% level of confidence for nitrogen and country significant at the 95% level for both nitrogen and moisture. Noting the modest significance of country effects, means by country of origin were calculated, and are shown in 8. The differences are all comparatively small: not more than 0.06g/100g for nitrogen and under 0.4g/100g for moisture.

Table 9 shows data for the nitrogen content by gender and size averaged across breed and country of origin. Again, as with the original 84 samples, heavy male birds differed significantly (p<0.001) in nitrogen content from the remaining three subsets. Comparing the averages shown in Table 9 with averages for breed in the 2000 study, values for females and light males are not significantly different from those for broilers in the 2000 study. However for heavy males, the values found here are appreciably lower (by 0.15 to 0.2 g/100g) than previously found either for broilers or for heavy broilers.

6. CONCLUSIONS

The UK market for chicken breast portions is obtained from both light and heavy broilers of both Ross and Cobb breeds. Although the Ross breed predominates in the UK market at the moment, this could change in the near future. It now is common practice not to separate male and female birds destined for the broiler market after hatching. Hence in arriving at mean values representative of the market, equal numbers of light, heavy, Ross and Cobb samples were taken. In addition, spent hen meat is no longer used for the manufacture of UK chicken products

The mean results of the chemical analyses on the population of 84 samples indicate that there has been a small reduction in fat content (1.5g/100g instead of 2.0 g/100g) and the hydroxyproline content


Table 8 Mean Moisture and Nitrogen (%) by Country of 144 samples

Country Nitrogen

Moisture

NL 3.73 74.65

PL 3.69 74.97

UK 3.67 74.65

Table 7 Nitrogen (%) by Breed, Size and Gender of 144 samples

Breed Size Gender LSMean SEM

Cobb H F 3.77 0.03

Ross H F 3.66 0.03

Cobb L F 3.76 0.03

Ross L F 3.74 0.03

Cobb H M 3.65 0.03

Ross H M 3.57 0.03

Cobb L M 3.71 0.03

Ross L M 3.72 0.03

Table 6 Moisture (%) by Breed, Size and Gender of 144 samples

Breed Size

Gender LSMean SEM

Cobb H F 74.19 0.14

Ross H F 75.01 0.14

Cobb L F 74.56 0.14

Ross L F 74.67 0.14

Cobb H M 74.99 0.14

Ross H M 75.25 0.14

Cobb L M 74.92 0.14

Ross L M 74.46 0.14

Table 9 Nitrogen (%) Averaged over Breed and Country of 144 samples

Size Gender LSMean SEMH F 3.71a 0.02L F 3.75a 0.02H M 3.61b 0.02L M 3.72a 0.02

a,b Statistically significant differences (p<0.01) were found between means a and b.

(0.05g/100g instead of 0.08g/100g) when compared to the 2000 study. The average nitrogen content for all broilers is also slightly down from the 2000 study as well (3.70g/100g instead of 3.78g/100g). The observation that the nitrogen content of breasts from heavy male broilers was significantly lower than the nitrogen contents from lighter birds and heavy females was unexpected.

In order to check that the above result was not anomalous, all of the nitrogen and moisture results obtained in the DG-AGRI project from all breast samples collected from the UK, Poland and the Netherlands (144 samples) were analysed statistically. Again, nitrogen values for breast samples from heavy male birds are significantly lower than those from light birds or heavy females. Taking equal numbers of heavy and light birds, the average nitrogen content calculated from Table 9 is 3.70g/100g.

7.0 RECOMMENDATION

In terms of the new values from the analyses of chicken breast samples, the average fat content of lean skinless chicken breast is 1.5g/100g, and the hydroxyproline content is 0.05g/100g.The average nitrogen content of lean skinless breast from all broilers is 3.70g/100g with a standard error of the mean of 0.02 (n=144). Where the raw material of a product is known to be skinless chicken breast, a fat-free nitrogen factor for skinless chicken breast of 3.75 is recommended. This value has been calculated from an average fat content of 1.5g/100g, rounded to the nearest 0.05 and compares to the recommended fat-free factor in the 2000 study for skinless chicken breast of 3.85, which represents a 3% difference. In any future study on chicken preparations, the recommended nitrogen factor of 3.75 should be used in conjunction with the methodology described in Commission Recommendation 175/20057.

8.0 ACKNOWLEDGEMENTS

This study was undertaken as part of the programme of work of the Royal Society of Chemistry, Analytical Methods Committee – Nitrogen Factors Sub-Committee. The project was part-funded by Defra as part of its Food Authenticity Programme, and partly by the Analytical Methods Trust. The European Commission - DG-AGRI is thanked for giving permission to use the samples and results from its Project 30-CE-0460798/00-25.

9. REFERENCES

1. The Food Labelling Regulations 1996 (SI 1996 No.1499), as amended, ISBN: 0-11-035941-0

2. Meat Products (England) Regulations 2003 (SI 2003 No. 2075), ISBN 0 11 047359 0)3. Council Regulation (EC) No 1234/2007 of 22 October 2007 establishing a common organisation of

agricultural markets and on specific provisions for certain agricultural products (Single CMO Regulation). OJ L299, 16.11.2007, p1-242.

4. Council Regulation (EC) No 1047/2009 of 19 October 2009 amending Regulation (EC) No 1234/2007 establishing a common organisation of agricultural markets as regards the marketing standards for poultrymeat. ) OJ L 290, 6.11.2009, p1-3.

5. Nitrogen Factors for Chicken Meat. Analyst, 2000, 125, 1359–1366

6. Food Standards Agency Survey of Meat Content, Added Water and Hydrolysed Protein In Catering Chicken Breasts (Number 20/01). http://tna.europarchive.org/20120530191353/http://www.food.gov.uk/science/surveillance/fsis2001/20chick

7. Commission Recommendation of 1 March 2005 concerning a Coordinated Programme


http://tna.europarchive.org/20120530191353/http://www.food.gov.uk/science/surveillance/fsis2001/20chick


for the Official Control of Foodstuffs for 2005, ANNEX V. OJ L59 05/03/2005, p 27-39

8. Study of the Physiological Water Content of Poultry Reared in the EU. Final Report 2012. http://ec.europa.eu/agriculture/external-studies/index_en.htm

9. Commission Regulation (EC) No 543/2008 of 16 June 2008 laying down detailed rules for the application of Council Regulation (EC) No 1234/2007 as regards the marketing standards for poultrymeat. OJ L157, 17.6.2008, p 46-87.

ANNEX 1Results of Analyses of 84 Chicken Breast Samples (g/100g)

Sample Number

Sample Description

Ash Mean Fat Mean HyP Mean Moisture

MeanNitrogen

Mean Total

M3003083 Ne_C_B - M_H 1.25 1.21 0.06 75.43 3.61 100.44


M3003468 Po_C_B - M_H 1.24 1.11 0.05 75.8 3.65 100.95M3003068 Ne_C_B - F_L 1.23 1.02 0.04 74.93 3.78 100.80M3002868 UK_C_B - M_H 1.33 1.26 0.06 74.33 3.85 100.98M3003419 Po_R_B - M_H 1.25 1.32 0.05 76.1 3.57 100.98M3003406 Po_R_B - F_L 1.29 1.17 0.05 75.7 3.71 101.34M3003021 Ne_R_B - F_L 1.32 1.54 0.04 75.03 3.80 101.64M3003394 Po_R_B - F_H 1.19 1.05 0.04 75.53 3.61 100.33M3003069 Ne_C_B - F_L 1.31 1.30 0.05 73.98 3.85 100.65M3003454 Po_C_B - F_L 1.33 1.35 0.04 73.97 3.84 100.65M3003082 Ne_C_B - F_H 1.29 1.32 0.06 74.23 3.88 101.08M3003429 Po_C_B - F_L 1.26 1.34 0.05 74.44 3.72 100.29M3002891 UK_C_B - M_H 1.17 2.32 0.05 76.2 3.36 100.69M3002881 UK_C_B - M_L 1.25 1.86 0.05 74.9 3.58 100.39M3003033 Ne_R_B - F_H 1.19 1.68 0.05 74.51 3.50 99.26M3003433 Po_C_B - M_L 1.29 1.60 0.06 75 3.67 100.83M3002854 UK_C_B - F_L 1.17 2.50 0.06 74.09 3.61 100.33M3003032 Ne_R_B - F_H 1.19 1.02 0.05 74.03 3.95 100.92M3002997 Ne_R_B - F_L 1.25 1.21 0.04 74.45 3.75 100.35M3003409 Po_R_B - M_L 1.28 0.81 0.05 74.97 3.67 100.00M3003070 Ne_C_B - F_L 1.27 1.06 0.05 74.4 3.79 100.42M3003396 Po_R_B - M_H 1.17 2.16 0.05 74.29 3.52 99.62M3002869 UK_C_B - M_H 1.24 1.33 0.04 74.7 3.88 101.52M3003011 Ne_R_B - M_H 1.19 1.06 0.04 75.63 3.73 101.18M3003430 Po_C_B - F_L 1.20 0.97 0.04 75.2 3.62 99.99M3003407 Po_R_B - M_L 1.31 1.45 0.05 74.37 3.80 100.88M3003395 Po_R_B - M_H 1.17 1.37 0.06 75.73 3.63 100.95M3003418 Po_R_B - F_H 1.27 1.71 0.04 74.23 3.81 101.02M3003008 Ne_R_B - F_H 1.29 1.19 0.04 75 3.87 101.66M3002808 UK_R_B - M_L 1.14 1.50 0.06 74.16 3.56 99.05M3003466 Po_C_B - F_H 1.19 1.35 0.05 74.84 3.49 99.19M3002840 UK_R_B - F_H 1.21 1.57 0.05 74.34 3.67 100.06M3003421 Po_R_B - M_H 1.13 1.58 0.04 76.86 3.33 100.38M3003023 Ne_R_B - M_L 1.27 1.18 0.04 74.88 3.73 100.64M3002820 UK_R_B - M_H 1.21 2.54 0.06 75.03 3.48 100.52M3002821 UK_R_B - M_H 1.29 1.53 0.05 73.84 3.76 100.16M3002999 Ne_R_B - M_L 1.26 1.86 0.06 73.8 3.68 99.92M3002841 UK_R_B - F_H 1.20 1.63 0.05 75.9 3.48 100.48M3003465 Po_C_B - F_H 1.21 1.21 0.04 74.4 3.74 100.19M3002866 UK_C_B - F_H 1.24 1.23 0.05 73.69 3.78 99.78M3003046 Ne_C_B - F_L 1.27 1.01 0.04 74.77 3.78 100.67M3003037 Ne_R_B - M_H 1.22 2.21 0.05 74.88 3.58 100.68M3003382 Po_R_B - F_L 1.27 1.26 0.07 74.59 3.76 100.62M3003020 Ne_R_B - F_L 1.27 0.90 0.05 74.63 3.80 100.55M3003057 Ne_C_B - F_H 1.30 1.42 0.06 73 4.00 100.72M3003416 Po_R_B - F_H 1.22 0.87 0.04 74.77 3.70 99.98M3002879 UK_C_B - M_L 1.23 1.55 0.05 73.45 3.88 100.48M3002880 UK_C_B - M_L 1.24 2.12 0.05 74.37 3.59 100.16M3003456 Po_C_B - M_L 1.24 1.41 0.05 75.06 3.72 100.96


M3002865 UK_C_B - F_H 1.24 0.96 0.04 74.97 3.83 101.10M3002829 UK_R_B - F_L 1.23 2.32 0.05 74.57 3.66 100.99M3003072 Ne_C_B - M_L 1.25 1.30 0.05 74.63 3.83 101.11M3003049 Ne_C_B - M_L 1.12 1.37 0.06 75.1 3.67 100.53M3003059 Ne_C_B - M_H 1.08 2.24 0.05 75.42 3.47 100.43M3002876 UK_C_B - F_L 1.21 1.53 0.05 75 3.73 101.05M3003431 Po_C_B - M_L 1.24 1.17 0.05 76.2 3.62 101.24M3003441 Po_C_B - F_H 1.20 1.72 0.04 74.77 3.75 101.13M3003405 Po_R_B - F_L 1.34 1.18 0.04 75.1 3.79 101.30M3002852 UK_C_B - F_L 1.29 1.88 0.05 73.87 3.80 100.79M3002817 UK_R_B - F_H 1.19 2.86 0.05 74.23 3.66 101.15M3002855 UK_C_B - M_L 1.26 1.01 0.06 75.27 3.69 100.60M3003432 Po_C_B - M_L 1.26 1.73 0.07 74.75 3.71 100.93M3003444 Po_C_B - M_H 1.28 1.82 0.06 74.66 3.73 101.07M3003381 Po_R_B - F_L 1.47 1.45 0.05 74.87 3.77 101.35M3002809 UK_R_B - M_L 1.29 1.29 0.04 74.2 3.83 100.71M3003080 Ne_C_B - F_H 1.31 1.69 0.05 73.87 3.89 101.18M3003061 Ne_C_B - M_H 1.15 2.45 0.05 75.13 3.45 100.29M3003025 Ne_R_B - M_L 1.21 1.46 0.05 74.68 3.62 99.97M3002831 UK_R_B - M_L 1.23 1.29 0.05 74.67 3.74 100.57M3002890 UK_C_B - F_H 1.31 1.02 0.04 74.2 3.80 100.28M3002819 UK_R_B - M_H 1.13 3.11 0.07 75.67 3.36 100.91M3003404 Po_R_B - F_L 1.34 1.80 0.04 74.48 3.74 101.00M3002747 UK_R_B - F_L 1.26 1.08 0.05 75.05 3.71 100.58M3002830 UK_R_B - F_L 1.27 1.43 0.05 74.63 3.74 100.70M3003000 Ne_R_B - M_L 1.28 1.35 0.04 74.1 3.91 101.16M3002893 UK_C_B - M_H 1.19 2.77 0.04 74.67 3.45 100.20M3003048 Ne_C_B - M_L 1.32 1.23 0.05 74.41 3.81 100.77M3003060 Ne_C_B - M_H 1.18 1.71 0.04 75.67 3.59 100.99M3002845 UK_R_B - M_H 1.26 1.50 0.06 75.39 3.71 101.34M3002832 UK_R_B - M_L 1.28 1.26 0.05 73.2 3.70 98.87M3003013 Ne_R_B - M_H 1.33 1.53 0.06 75.13 3.58 100.37M3003445 Po_C_B - M_H 1.23 1.72 0.06 75.13 3.68 101.09M3003442 Po_C_B - F_H 1.25 1.43 0.03 74.4 3.69 100.14M3003009 Ne_R_B - F_H 1.17 1.45 0.04 73.93 3.77 100.11

ANNEX 2

Figure 2 Proximate Values of All Replicates (in % m/m) Plotted Against One Another (Country of Origin: White=NL, Red=PL, Blue=UK)


Moisture

3.3 3.5 3.7 3.9 1.1 1.2 1.3 1.4

7374

7576

77

3.3

3.5

3.7

3.9

Nitrogen

Fat

1.0

2.0

3.0

73 74 75 76 77

1.1

1.2

1.3

1.4

1.0 1.5 2.0 2.5 3.0

Ash

Proximates (% by mass)

Figure 3 Mean Values (%) of Moisture, Nitrogen, Fat, Ash and Hydroxyproline by Breed

Cob

b

Ros

s

73

74

75

76

77

g/10

0g

Moisture

Cob

b

Ros

s

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

Cob

b

Ros

s

1.0

1.5

2.0

2.5

3.0

g/10

0g

Fat

Cob

b

Ros

s

1.1

1.2

1.3

1.4

g/10

0g

Ash

Cob

b

Ros

s

0.04

0.05

0.06

0.07

g/10

0g

Hydroxyproline

Breed

Figure 4 Mean Values (%) of Moisture, Nitrogen, Fat, Ash and Hydroxyproline by Country


NL PL UK

73

74

75

76

77g/

100g

Moisture

NL PL UK

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

NL PL UK

1.0

1.5

2.0

2.5

3.0

g/10

0g

Fat

NL PL UK

1.1

1.2

1.3

1.4

g/10

0g

Ash

NL PL UK

0.04

0.05

0.06

0.07

g/10

0g

Hydroxyproline

Country

Figure 5 Mean Values (%) of Moisture, Nitrogen, Fat, Ash and Hydroxyproline by Gender

F M

73

74

75

76

77

g/10

0g

Moisture

F M

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

F M

1.0

1.5

2.0

2.5

3.0

g/10

0g

Fat

F M

1.1

1.2

1.3

1.4

g/10

0g

Ash

F M

0.04

0.05

0.06

0.07

g/10

0g

Hydroxyproline

Gender

Figure 6 Mean Values (%) of Moisture, Nitrogen, Fat, Ash and Hydroxyproline by Size (L,H)

L H

73

74

75

76

77

g/10

0g

Moisture

L H

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

L H

1.0

1.5

2.0

2.5

3.0

g/10

0g

Fat

L H

1.1

1.2

1.3

1.4

g/10

0g

Ash

L H

0.04

0.05

0.06

0.07

g/10

0g

Hydroxyproline

Size

ANNEX 3


3.3 3.4 3.5 3.6 3.7 3.8 3.9 4.0

73

74

75

76

77

Nitrogen

M oi st ur e

NLPLUK

Figure 7 Mean Nitrogen and Moisture Contents (%) for 144 Samples by Country

Figure 8 Moisture and Nitrogen Contents (%) Figure 9 Moisture and Nitrogen Contents (%)by Gender (M,F) by Size (L,H)


F M

73

74

75

76

77

g/10

0g

Moisture

F M

3.3

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

Gender

H L

73

74

75

76

77

g/10

0g

Moisture

H L

3.3

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

Size

Figure 10 Moisture and Nitrogen Contents (%) Figure 11 Moisture and Nitrogen Contents (%)by Breed (Ross and Cobb) by Country (PL, NL, UK)


Cob

b

Ros

s

73

74

75

76

77

g/10

0g

Moisture

Cob

b

Ros

s

3.3

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

Breed

NL

PL

UK

73

74

75

76

77

g/10

0g

Moisture

NL

PL

UK

3.3

3.4

3.5

3.6

3.7

3.8

3.9

4.0

g/10

0g

Nitrogen

Country

References to published material9. This section should be used to record links (hypertext links where possible) or references to other

published material generated by, or relating to this project.

REFERENCES FROM REPORT

1. The Food Labelling Regulations 1996 (SI 1996 No.1499), as amended, ISBN: 0-11-035941-0

2. Meat Products (England) Regulations 2003 (SI 2003 No. 2075), ISBN 0 11 047359 0)

3. Council Regulation (EC) No 1234/2007 of 22 October 2007 establishing a common organisation of agricultural markets and on specific provisions for certain agricultural products (Single CMO Regulation). OJ L299, 16.11.2007, p1-242.

4. Council Regulation (EC) No 1047/2009 of 19 October 2009 amending Regulation (EC) No 1234/2007 establishing a common organisation of agricultural markets as regards the marketing standards for poultrymeat. ) OJ L 290, 6.11.2009, p1-3.

5. Nitrogen Factors for Chicken Meat. Analyst, 2000, 125, 1359–1366

6. Food Standards Agency Survey of Meat Content, Added Water and Hydrolysed Protein In Catering Chicken Breasts (Number 20/01). http://tna.europarchive.org/20120530191353/http://www.food.gov.uk/science/surveillance/fsis2001/20chick

7. Commission Recommendation of 1 March 2005 concerning a Coordinated Programme for the Official Control of Foodstuffs for 2005, ANNEX V. OJ L59 05/03/2005, p 27-39

8. Study of the Physiological Water Content of Poultry Reared in the EU. Final Report 2012. http://ec.europa.eu/agriculture/external-studies/index_en.htm

9. Commission Regulation (EC) No 543/2008 of 16 June 2008 laying down detailed rules for the application of Council Regulation (EC) No 1234/2007 as regards the marketing standards for poultrymeat. OJ L157, 17.6.2008, p 46-87.




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