The document discusses the role of carotenoids, specifically Carophyll® red and yellow, in the pigmentation of chicken and egg yolks, emphasizing the importance of dietary carotenoids for achieving desired coloration. It outlines various carotenoids and their sources, as well as the methods for evaluating pigmentation in poultry. Additionally, it highlights the stability and quality characteristics of Carophyll® products compared to other sources of carotenoids in the animal nutrition market.

1. FOR INTERNAL USE ONLY
CAROPHYLL® Red and Yellow for
Chicken and Egg Pigmentation
VMS, 2013
F. Cisneros

2. FOR INTERNAL USE ONLY
Content:
•
•
•
•
•
Bright Food and carotenoids
The pigmentation process
Egg pigmentation
Chicken pigmentation
The CAROPHYLL® brand

3. FOR INTERNAL USE ONLY
Yolk Color Preferences in Various Countries, DSM
Yolk Color Fan(DYCF)
100%
90%
80%
70%
DSM 6
60%
DSM 8
DSM 10
50%
DSM 12
40%
DSM 13≤
30%
20%
10%
England
France
Italy
Spain
Germany
India
S. Korea
Russia
Australia
Brazil
China
0%

4. FOR INTERNAL USE ONLY
Broiler pigmentation in different markets

5. FOR INTERNAL USE ONLY
The color of the egg yolk and chicken skin is determined by the concentration
and type of carotenoids in the diet, as well as their absorption and overall
ingestion.
A pigmented egg always comes from a healthy hen.

6. FOR INTERNAL USE ONLY
Colors exist in nature due to the presence and combination of different
groups of molecules such as chlorophyll, quercetin, and carotenoids

7. FOR INTERNAL USE ONLY
• 1831 β-Carotene from carrots by Wackenroder
• Structure by Kerrer in 1928-1931
• Synthesis by Kerrer and Inhoffen in 1950

8. FOR INTERNAL USE ONLY
b-Carotene
400 to 600 nm

9. FOR INTERNAL USE ONLY
b-Carotene
O
OH
O
HO
Canthaxanthin
Zeaxanthin
O
OH
HO
O
Astaxanthin
O
OH
HO
O
Lutein

10. FOR INTERNAL USE ONLY
Commonly Used Terms for
Carotenoids
Lutein
Zeaxanthin
Cryptoxanthin
Violaxanthin
Canthaxanthin
Apo-ester
Citranaxanthin
Capsanthin
Capsorubin
Astaxanthin
Carotenes
Xanthophylls
Xanthophylls
or
Oxy-carotenoids
Terpene compounds
that, due to their
structure, selectively
absorb light
Carotenoids

11. FOR INTERNAL USE ONLY
Main Carotenoids in Poultry Pigmentation
Apo-ester
Lutein
Canthaxanthin
Zeaxanthin
445
450
Yellow Carotenoids
455
460
465
470
475
480
Wave lenght, nm
Adapted from Bauernfeind, 1981 and Schierle 2011

12. FOR INTERNAL USE ONLY
Pigmentation Level
Saturation and Color Phases
Lutein / zeaxanthin
Apo-ester
Canthaxanthin
Carotenoids Present in Feed
Technical guidelines mention a combination of yellows and reds as the most efficient way to
reach the target color and homogeneity

13. FOR INTERNAL USE ONLY
Egg Pigmentation
Sources of Carotenoids
Pigmentation Evaluation

14. FOR INTERNAL USE ONLY
Main Carotenoids in Egg Yolk:
•Canthaxanthin
•Capsanthin/Capsorubin
•Citranaxanthin
•Apo-ester
•Lutein/Zeaxanthin

15. FOR INTERNAL USE ONLY
%
Efficacy compared to canthaxanthin, %
Relative Efficiency of Paprika Extract
Products (Canthaxanthin*=100%)
target DSM YCF
45
12.9
40
14.4
35
30
25
20
15
P1
P2
P3
P4
P5
P6
P7
P8
*One g of Canthaxanthin from Carophyll red is equivalent to 2.4 to 6 g of paprika extracts (19 -142 g of commercial paprika products), depending on source
and target yolk color.
Fuente: HUYGHEBAERT, G. (2008) The utilisation of red oxy-carotenoids from paprika extracts for egg yolk pigmentation. 23rd WPC, in Brisbane, Australia

16. FOR INTERNAL USE ONLY
Carotenoid Makeup of Commercial
Paprika Extracts
100%
80%
Cap/Cap
Citranaxanthin
60%
Zeaxanthin
Lutein
40%
20%
0%
1
2
3
4
5
Source: HUYGHEBAERT, G. (2008) The utilisation of red oxy-carotenoids from paprika extracts for egg yolk pigmentation.
23rd WPC, in Brisbane, Australia

17. FOR INTERNAL USE ONLY
Citranaxanthin in Egg Yolk Pigmentation
15
RYCF
14
A poE 3 ppm
13
Lut / Ze a 9 ppm
12
11
10
Red Carotenoids (ppm)
Source: Adapted from Huyghebaert Gérard1 Piesschaert Isabelle. 4ème Journées de la Recherche Avicole, Nantes 27-29 mars 1991

18. FOR INTERNAL USE ONLY
DSMYCF
Effect of the Source of Yellow Carotenoid on
Egg Yolk Pigmentation
2011, France
1991, Mexico
9
RYCF
Apo-ester
Tagetes
0
Apo-ester
8
13
12
11
10
9
8
7
6
5
4
3
2
Tagetes
2
4
6
8
10 12 14 16 18 20
Yellow Carotenoids Present in Feed, ppm
7
6
3 g of tagetes equal 1 g of
Apo-ester
5
0
2
4
6
8
10
12
Carotenoids Present in Feed, ppm
Source:Vallejo, Avila & Tirado, 1991. proceeding AMENA meeting. Ixtapa, Mexico
Source: Faruk, Aurelli and Schierle. 2011. CRNA Report 00010749. France

19. FOR INTERNAL USE ONLY
Boiled Eggs

20. FOR INTERNAL USE ONLY
Effect of the Source of Canthaxanthin on
the Yolks of Fresh and Boiled Eggs
DSM Yolk Color Fan
14
a
b
b
12
b
c
10
a
b
8
CR
b
C1
c c
6
C2
4
C3
2
C4
0
Fresh
Boiled
Source: Aureli, et al. 2010. Efficiency of various red carotenoids
and their in-yolk stability after boiling. EPC. Tours, France. 2010

21. FOR INTERNAL USE ONLY
Egg Pigmentation
Sources of Carotenoids
Pigmentation Evaluation

22. FOR INTERNAL USE ONLY
Comparison of Product Forms
Equivalent of 1 kg of pure Canthaxanthin to extracts of
paprika as sources of red carotenoids for Egg
pigmentation
800
700
700
600
600
500
500
400
300
200
100
0
Page
1
10
2.5

23. FOR INTERNAL USE ONLY
Comparison of Product Forms
Equivalent of 1 kg of pure Apo-ester from different
sources for Egg pigmentation
250
210
200
150
150
100
50
0
1
10
3
165
180

24. FOR INTERNAL USE ONLY
The DSM Yolk Color Fan
Pros: Standard, single measurement, inexpensive, and easy to use
Cons: Subjective, variations among individuals

25. FOR INTERNAL USE ONLY
Colorimeters: L*a*b* and L*C*h*
Color Spaces
Pros: Objective, repeatability, automatic, portable
Cons: Three different measurements, non-standard, higher price
https://wwws.konicaminolta.net/php/global/se_booklet_download/entry.php

26. FOR INTERNAL USE ONLY
Automatic Measuring Devices
http://www.tss-york.com/products/qcmrange#QCC
ORKA FOOD TECHNOLOGY
http://eggtester.com/products/egg-analyzer

27. FOR INTERNAL USE ONLY
Industrial Eggs

28. FOR INTERNAL USE ONLY
Effect of Dietary Apo-ester on Egg Yolk
Concentration and Color
14
100
Minolta Yellow
90
12
80
DSM Yolk Color Fan
10
70
60
8
50
6
40
30
4
20
Yolk Apo-ester, ppm
2
10
0
0
0
5
10
Dietary Apo-ester, ppm
CRNA RPD 00010749 DSM internal, 2012
20

29. FOR INTERNAL USE ONLY
Effect of Dietary Carotenoids on Egg Yolk
Concentration and Color in Hy-Line W-36
14
12
60
L* a* b* Yellow
50
DSM Yolk Color Fan
10
40
8
30
6
20
Total Egg Carotenoids, ppm
4
10
2
0
0
3.4
26.7
37.9
62.3
Carotenoids Present in Feed, ppm
Carlos de la Cruz, 2006
97.9
128.2

30. FOR INTERNAL USE ONLY
Effect of Two Sources of Carotenoids on the Concentration of
Carotenoids and Minolta Yellow Units in the Whole Egg
110
110
100
100
90
90
BCE, ppm
BCE, ppm,
BCE, ppm
80
70
Minolta Yellow
80
70
BCE, ppm
Minolta Yellow
60
60
50
50
40
40
60
80
40
120
140
160
180
200
220
Dietary Lutein/Zeaxanthin, ppm
Dietary Apo-ester, ppm
•Color is not a good indicator of carotenoid concentration in egg yolk
•Apo-ester can linearly increase carotenoid concentration in egg yolk
Sirri, 2007. JAPR, 16:249
240

31. FOR INTERNAL USE ONLY
HPLC

32. FOR INTERNAL USE ONLY
i-check
Schweigert,F. et al. 2010. XIII European Poultry Conference.Tours,France.

33. FOR INTERNAL USE ONLY
Chicken Pigmentation
Sources of Carotenoids
Pigmentation Evaluation

34. FOR INTERNAL USE ONLY
Main Carotenoids in Chicken Pigmentation:
•Canthaxanthin
•Capsantin/Capsorubin
•Citranaxanthin
•Apo-ester
•Lutein/Zeaxanthin

35. FOR INTERNAL USE ONLY
Apo-ester to Tagetes Equivalency
Trial details
•
416 Ross PM3 chickens, 4 treatments containing 5ppm of red base
(Canthaxanthin) for all treatments
•
Added xanthophylls according to analytical values (below average tagetes),
43% average
T1
T2
T3
T4
LZ/APE (ppm)
60/0
50/4
40/8
20/16
a (L*a*b*)
3.2a
3.6a
4.1b
4.2b
b (L*a*b*)
38a
36.3b
35.8b
31.3c
DSM Broiler Fan
103.7a
103.5a
103.6a
102.8b
a, b, c: significant differences in the same line ( P<0.001)
Source: Hamelin and Hernandez, XIIIth European Poultry Conference, Tours, 23-27 August 2010

36. FOR INTERNAL USE ONLY
Apo-ester to Tagetes Equivalency
Protocol:
•
•
•
•
•
•
Spanish Trial Farm
September-October 2011
Ross 308 in balanced mixed groups
Two pelleted meals (0-12 and 22-32 days)
Wheat, soybean meal, corn, barley, sorghum
240 chickens were used for the carcass color evaluation (balanced by
sex), 60 per treatment
Source: Hamelin, Martinez y Martinez. 2012. Memorias Simposio Cientifico de Avicultura. WPSAAECA, Barcelona.

37. FOR INTERNAL USE ONLY
Apo-ester to
Tagetes
Equivalency
Canthaxanthin
(ppm)
Apo-ester
(ppm)
Lutein +
Zeaxanthin (ppm)
5
0
30
5
20
0
30
5
20
T1
T2
T3
10
T4
DSM Broiler Fan
105.5
105.1
105
104
103.7
31
31.8
30.7
30.3
30
29
103.5
103
32
32
104.6
104.3
104.5
Minolta, b*
33
28
a
b
NS NS
27
102.5
26
102
25
5
10
Canthaxanthin, ppm
30 20/5
Lut-Zea/Apo-ester, ppm
a b
5
NS NS
10
30 20/5
Canthaxanthin, ppm
Lut-Zea/Apo-ester, ppm
Source: Hamelin, Martinez y Martinez. 2012. Memorias Simposio Cientifico de Avicultura. WPSAAECA, Barcelona.

38. FOR INTERNAL USE ONLY
High in Zeaxanthin
Analyzed Yellow Xanthophylls (XA)
Lutein
Zeaxanthin
T-1
30
26
25
22
24
4.4
3.4
T-2
35 ppm XA (10% Zea) + 5 ppm Cantha
34
8.5
T-3
b* Minolta Skin
No XA
40 ppm XA (25% Zea)
25
14.2
T-4
32 ppm XA (10% Zea) + 2 ppm Cantha
31
6.8
20
Deposition Rate in Skin, %
15
10
5
0
Lutein
T-2
d
T1
a
T2
c
T3
b
T4
Source: Adapted from P Vendrell, et al., 2001. Poult. Sci 80:320-326.
.
12
10
T-3
9
Zeaxanthin
9
8
T-4
12
9

39. FOR INTERNAL USE ONLY
Chicken Pigmentation with Tagetes, Apo-ester and
Canthaxanthin
Lutein/Zeaxanthin
(ppm)
Lutein/Zeaxanthin- Apo-esterCanthaxanthin
Grower
80
65 + 4 + 4
Finisher
90
75 + 4 + 4
18%
Lutein/Zea, Apo, Cantha
15%
Lutein/Zea
12%
9%
6%
3%
0%
26 28 30 32 34 36 38 40 42 44 46 48 50 52 54 56
b de L*a*b*
S. Fernandez, 2010. W. Nutrition Conference. Sask. Canada.

40. FOR INTERNAL USE ONLY
Chicken Pigmentation
Sources of Carotenoids
Pigmentation Evaluation

41. FOR INTERNAL USE ONLY
Pigmentation Evaluation
-
In order to establish the range of colors, chromatometer measurements were taken of
chicken skins with various degrees of pigmentation
-
The fan’s material absorbs brightness, thus reducing variability
-
Each blade is designed to include a viewing area to better compare different chicken
samples
-
The device is valid in Europe, South America, and Asia

42. FOR INTERNAL USE ONLY
Pigmentation Evaluation
Methodologies Used
• Minolta
• HPLC (concentration in the skin)
• i-check (concentration in the skin)
• Beta-Carotene equivalents in plasma
• Yolk Color Fan

43. FOR INTERNAL USE ONLY
Carophyll® is the highest quality, best recognized brand of carotenoids in
the animal nutrition market
Carophyll® red 10% is a source of the carotenoid
canthaxanthin, identical to the one found in
nature in Chanterele Mushrooms
Carophyll® yellow 10% is a source of the
carotenoid Apo-ester, a sterified version of
apocarotenoic acid, a widely distributed
carotenoid found in grass, alfalfa and other green
plants

44. FOR INTERNAL USE ONLY
CAROPHYLL® Characteristics:
• The best stability during the feed process in the market today
• The best uniformity
• The only sources of Canthaxanthin and Apo-ester which are animal by
product-free, so it can used in Kosher and Halal programs and well
suited breeder programs
• The only sources of Canthaxanthin or Apo-ester which are water
dispersible, allowing the reinforcing of any pigmentation program in
the water
• CAROPHYLL® is produced in France along the Rhine River and subject
to the highest environmental standards of Europe
• No recall has been ever activated for a bag of CAROPHYLL® product,
as long as our records can detect (over thirty years)

45. FOR INTERNAL USE ONLY
Stability of CAROPHYLL® Red 10% compared to
European sources of Canthaxanthin along the
feed process for 30 days
110%
Retention (%)
100%
CAROPHYLL Red 10%
90%
CAROPHYLL Red 10%
CAROPHYLL Red 10% Avg
Canthacol 10%
80%
Canthacol 10%
Canthacol 10% Avg
70%
Lucantin Red
Lucantin Forte
60%
50%
initial value
Pmx 30 days
Pelleting 90C
Feed 30 days
CAROPHYLL® Red showed its superior performance in premix and feed stability
Samples stored at 30 °C and 65% humidity in polyethylene (air permeable) bags. Duplicate analysis.
Three batches of CR and four batches of Canthaplus tested.
Trial VFP201234, VFP201132, VFP29036

46. FOR INTERNAL USE ONLY
Stability of CAROPHYLL® Red 10% compared to
different sources of Canthaxanthin along the
feed process for 30 days
100%
CAROPHYLL Red 10%
Retention (%)
90%
CAROPHYLL Red 10%
CAROPHYLL Red 10% Avg
CHNA
80%
CHNB
CHNC
CHND
70%
CHNE
CHNF
CHNG
60%
CHNH
Chinese Avg
50%
initial value
Pmx 30 days
Pelleting 90C
Feed 30 days
Chinese sources of Canthaxanthin delivered variable and mostly poor performances
when compared to CAROPHYLL® Red 10%
Samples stored at 30 °C and 65% humidity in polyethylene (air permeable) bags. Duplicate analysis.
Trial VFP201234, VFP201132, VFP29036

47. FOR INTERNAL USE ONLY
Retention (%)
82.7%
ApoEplus can see their Apo-ester degraded at a rate up to 0.7% for every storage day.
Therefore, even a week in premix or in the bin will make a difference.
Samples stored at 30 °C and 65% humidity in polyethylene (air permeable) bags. Two samples
per batch, duplicate analysis. Two batches of CY and four batches of ApoEPlus. Same feed and
premix formulation. Pelleting 90°C, 20 s retention.
Trials VFP201252, VFP201262

48. FOR INTERNAL USE ONLY
Great variability among different marigold products, even from the same brand. Low stability in most of them.
Low variability with Carophyll® Yellow, best performances, consistent results.
Samples stored at 30 °C and 65% humidity in polyethylene (air permeable) bags. Two samples
per batch, duplicate analysis. Two batches of CY . Same feed and premix formulation.
Conditioning time 20 S .
Trials VFP201252, VFP201262