1. EUROTIOX

2. Feed Ingredients Vitamins, pigments Fat, oils (energy)
OXIDATION
Feed Ingredients
Vitamins, pigments
Fat, oils (energy)

3. AUTOXIDATION SERIES OF DESTRUCTIVE CHEMICAL PROCESSES
OCCURS IN ORGANIC MATERIALS
CHANGES THE CHEMICAL NATURE OF AFFECTED MOLECULES
INCREASINGLY FAST
ROOM TEMPERATURE REACTION
CAUSES RANCIDITY IN FATS, OILS
Series of destructive chemical process, it is important to mention that it is also irreversible.
This reaction occurs mainly in organic materials
Due to the reaction, the chemical nature of molecules is changed. Vitamins for example will loose their activity.
The auto oxydation is an increasingly fast reaction, which is already present within a few hours after mixing. In untreated raw materials, often the pericarp protects the material from oxydation (no oxygen), once it is grinded, protection has dissappeared.
The reaction of autooxydation is a reaction which takes place at room temperature, this in contradiction to many other chemical reactions. Of coarse higher temperatures will increase the speed of the reaction.
BIOTIN

4. Oxidation influenced by:
Moisture: free water in raw material and feed
Catalysts: metals promoting free radical formation,
Light: promotes oxidation
Temperature: increase oxidation also during cooling
Contact with air: improves peroxide formation
Enzymes: lipase, lipoxygenase
Time: storage time, production
Complet reactions involved including all of the above
Moisture is outside (humidity) also Aw, Humidity from steam during the process.
Cataly mostly in premixes

5. Aldehydes from Fat Autoxidation
Frankel, 1984

6. THE INFLUENCE OF OXIDISED FAT ON THE MICRO-FLORA IN BROILERS (11 DAYS)

7. TOXIC METABOLITES OF FAT OXIDATION
Shorten small intestine villi length
FROM: Poul. Sci. Abstracts (1994) Vol. 73, Suppl. 1, pg 47.
Reduce nutrient adsorption

8. Autoxidation and Animal production
Reduced Egg production in laying hens (72%) (Voreck & Kirehgessner, 1981)
Autoxidation products are cancernogenic (Morita et al., 1983)
Even low increase in Peroxide Value (>7 meq/kg) has a significant effect on Feed Conversion Ratio (Cabel et al., 1988)
Rancid Feed has an impact on the gastro-intestinal Structure, Cell proliferation and intestinal Microflora (Shermer et al., 1995)
Oxidised vegetable oil in Broiler feed reduces growth rate (5%) (Engberg et al 1996)
Rancid fish oil induced anaemia in Mink, reduced iron absorption (Borsting et al., 1994)

9. Effect of peroxides on broiler performances (49 days) University of Arkansas% c a ab b %
Cabel, M. C., P. W. Waldroup, W. D. SHermer and D. F. Calabotta (1989), Poultry Science 67:

10. Effect of raw material quality on broiler performances (56 days) INTA (Argentina)% a a a b a a % a b
Schang et al

11. The three stages of OXIDATION
1) Initiation phase
RH > R° H°
Fat molecule
Free radical
2) Propagation phase
R° O > ROO°
ROO° RH > ROOH R°
(peroxide)
3) Termination phase
ROO = peroxide= faster oxidation
Aim is to control the peroxide levels so want to stay in initial phase
peroxide is only an intermediate molecule
peroxide good indicator of oxidation /quality……. Snap shot only
ROO° + ROO° ---> stable end products
(aldehydes, ketones, short chain fatty acids)

12. Oxidation Process involving Oxygen & lipids
temp.
Catalysts
Light
Air
Time
FREE
RADICALS
Damage critical
nutrients
Lipids
FREE
RADICALS
PEROXIDES +
Oxygen forms
Reaction with cell membranes
Toxic for animals
Destruction of fat soluble vitamins
Reduction of performances
PEROXIDES
Aldehydes
Ketons
Rancidity
Reduction of feed intake
En-products absorbed in the body
Start of in vivo oxidation

13. The three stages of OXIDATION
Peroxides value (meq)
Polimers, oxidised
fat, acids, aldehides
ketones
Peroxides, radicals
water
Therefore need to treat early because everything lost cannot be reversed, easier to stop/control reaction when reaction is slow
Induction
When there is dis -association of fat molecules and then reaction with oxygen
Oxidation Phase
dis-association + direct reaction from peroxide on fat molecule
Polimers, oxided fat, etc are all stable forms of lower energy value, also palatability issues
Induction Oxidation Termination period

14. Different antioxidants
Free radical scavengers:
Ethoxyquin, BHA, BHT, Propyl Gallate
Chelators:
Citric acid, phosphoric acid
Combination of 2 Free radical scavengers best (except in aquaculture)
Oxidation scavengers
to absorb oxygen from air not really revealent for feed
Chelators
To bind metals so not so availableto promote free radicals but not bound for nutrition. Only use citric acid as literature good chelator , we know that phosphoric acid sometimes works sometimes increases sometimes no effect (Endox Kemin uses phosphoric..is a cheeper ingredient)

15. Differences between antioxidants
Ethoxyquin:
very stable, only feed grade, raw materials
Gallates:
very stable, regenerates ethoxyquin or BHA activity
BHA:
heat stable, mainly food and petfood
BHT:
heat stability problem, repartition problem,
readily broken down in premixes

16. Structures of commonly used synthetic antioxidants
Butylated hydroxytoluene (BHT) Mol. Wt. 220
Tertiary butylhydroquinone (TBHQ) Mol. Wt. 166
Non allowed in UE
Ethoxyquin Mol. Wt. 217
These all are relatively short chain compounds compared to fats and Vitamins
Longer chain > reactivity and easier to oxidise (they are less stable)
> number of bonds > reactivity and easier to oxidise (again less stable)
When RH  RO - + H+ The fat chain then has a -ve charge which for reasons that we don’t really understand makes it easier for O2 to attach and oxidise the molecule leading to the reduction in nutrient value ransidity etc.
To stop this reaction anti-oxidant molecules (which are shorter chain so more stable) donate a H+ to reverse the Initial phase reaction this only helps DELAY oxidation as the RH  RO - + H+ is a dynamic reaction.
Propyl gallate Mol. Wt. 212
Butylated hydroxyanisole (BHA) Mol. Wt. 180

17. Triple Mode of Action
EUROTIOX
1.Citric acid forms reversible bonds with metal pro-oxidants
in the premix and feed to inhibit them from initiating
oxidation
2. Ethoxyquin blocks oxidation by donating hydrogen ions
to free and peroxy radicals formed during the onset
and propagation stages
3. Propyl gallate restores and potentiates ethoxyquin activity
up to 3 times by donating hydrogen ions from its 3 active
sites to oxidised (spent) ethoxyquin molecules

18. Efficacy of synthetic antioxidants
BHT <ETQ<BHA<gallate($30) for cost of material Gallate 20x more expensive than BHT
Data above is independent work
Slide to show why we use our combination of ETQ and Galate
Days
Stability test in mixed fat - Dr. M. Coelho (1995)

19. Formation of Ellagic Acid from Propyl Gallate

20. Antioxidant Activity of BHA, Propyl Gallate and Heterodimers
T. Kurechi and A.Kunugi, J. American Oil Chem. Soc. 60:33 (1983)

21. Ethoxyquin/Propyl Gallate SYNERGY mechanism

22. Ethoxyquin/Propyl Gallate SYNERGY mechanism

23. Effect of citric acid on soya oil stability
Metal chelating
Effect of citric acid on soya oil stability
Trail was done by adding metals to oil
With citric reduced oxidation so less peroxide
Mustakas et Al. (1984) 17

24. Inhibition of Initiation by Chelation of Copper
OOC
CH2 C
COO OH O HO
Cu++
+ Cu++
HOCCOOH
CH2COOH
Citric Acid
Cu Citrate complex with divalent cation, forming two five-membered rings

25. Eurotiox 32-Premix Comparative granulometry
Physical characteristics VIP after getting the right ingredients and ratios
because citric cannot use limestone as some other competitors are doing
need small homog. Particles, cannot be re-active with other ingredients
Particle size class ( Mesh, mm) - Sieve analysis by industrial lab, Belgium, July 1996

26. EUROTIOX unique antioxidant complex
ETHOXYQUIN
- high purity (% mono)
- high stability
PROPYL GALLATE
-highly effective
- EtQ regenarator
Two very potent free radical scaven-gers
Only
proven
synergism
Very
effective
economical
complex
stable
micro-
pearled +
CITRIC ACID
-metal scavenger
Maximum
protection
at low
dosage +
UNIQUE CARRIER
- high absorption
- free-flowing
- impr. stability +
ALUMINIUM bag
-protected 23 20

27. EUROTIOX ANTIOXIDANT PROGRAM
Vitamins, Pigments
protection
in premixes & feed
Preventing
rancidity in
fat, oils, bone
meals
EUROTIOX
32-Premix
EUROTIOX
L-32

28. Dosage in feed and raw materials
EUROTIOX
Dosage in feed and raw materials

29. EUROTIOX 32 Premix in feed
Eurotiox 32 Premix at g/t in Feed with < 4% fat and < 1 month storage time
+ 5 g/t for each 2% fat increase
+ 5 g/t for each month storage increase

30. EUROTIOX 32 Premix in feed

31. EUROTIOX 32 Premix in premixes and concentrates
Vitamin premix: 50g/ton
Vitamin and traces premix: 100 g/ton
Vitamin and traces and choline chloride: 200 g/ton
Higher dosages are required in case proteins are present.

32. EUROTIOX L 32 for raw material protection

33. EUROTIOX L 32 for raw material protection

34. Dosage of EUROTIOX L 32 Raw material
For one month protection
For Two Months Protection
For Three Months Protection
For Four Months Protection
Vegetable oil (crude) 75
150
200
300
Palm oil (crude)
Rice Bran oil (crude)
Rice Bran oil
(Refined)
Sunflower oil
(crude)
100
400
Copra oil (crude)
Soya oil (crude)
Fish oil (crude)
Poultry fat
Lard
Tallow 35 70
135
Mixed fat
MBM 30 60 90
120
Rice Bran
(14-16% Fat)
Full Fat Soy Bean
Source: communication AVITECH

35. Vitamin recovery test against oxidation (Vitamin A in a vitamin mineral premix) HPLC assay - 6 months testing period March ‘95-Sep ‘95
EUROTIOX 32-Premix - 45 g/ton
45 g/t EUROTIOX 32
200, g/t ETQ-66%
Control
Verlies van 450 naar 300 is toch nog steeds hoog, vandaar dosering naar 100 g. Wel blijkt duidelijk dat 45 g/Eurotiox vergelijkbaar is met 200 g Ethoxyquin.

36. Vitamin recovery test against oxidation (Vitamin A and E in a Broiler premix) HPLC assay - 3 months testing period - Netherlands - Oct ‘96/ Feb ‘97
EUROTIOX 32-Premix - 20 g/ton
89.4% Vitamin E
recovered after 3 months
94.5% Vitamin A
recovered after 3 months
Periode beperkt over 3 maanden, dit is in praktijk dikwijls te kort.
No significant loss of Vitamin A & E during a 3 month period
Vitamin A & E analytical detection test by HPLC performed by an independent
laboratory (SERLABO) in the Netherlands.

37. Vitamin A Stability in Premix Eurotiox 32 premix
EUROTIOX 32-Premix - 20 and 30 g/ton
Periode van 1 maand, in praktijk te weinig
Independant laboratory - HPLC analysis.

38. Vitamin A Stability comparative test Broiler feed - March 1997
EUROTIOX 32-Premix
At 25 g/ton, EUROTIOX provides the best VITAMIN A protection of all products tested
Project with large industrial company(Broiler integration) in Philippines. Vitamin detection
analysis performed by an independent laboratory (SERLABO) in the Netherlands.

39. Vitamin recovery test (Vitamin A in a vitamin mineral premix) HPLC assay - Broiler premix from Thailand - Independent laboratory Holland - Sep 99
EUROTIOX 32-Premix - 20 g/ton
Pdt N
125 g/ton
EUROTIOX
20 g/ton
Control
Pdt K
125 g/ton
Periode van 14 dagen, in praktijk te weinig

40. Trial results

41. EUROTIOX 32 PREMIX vs ENDOX
TRIAL AO 001

42. Eurotiox 32 Premix in meat and bone meal (poultry origin)
Trial set-up:
A control sample (no anti-oxydant added) and a treated sample (150 ppm Eurotiox 32) was incubated in the OXIPRESS.
The OXIPRESS was running until the control sample was completely oxidised.
Both samples were analysed on their nutritional value and compared to a control sample which was not put in the OXIPRESS at all.

43. Eurotiox 32 Premix in meat and bone meal (poultry origin)
CONTROL WITHOUT OXIPRESS
CONTROL WITH OXIPRESS
CONTROL WITH OXIPRESS PPM EUROTIOX 32 PREMIX
Crude fat (g/kg)
121.2
105.4
115.6
Metaboliseble Energy (MJ/kg)
11.13
10.22
10.95

44. Eurotiox 32 Premix in meat and bone meal (poultry origin)
-1.6%
-8.1%

45. OXIPRESS RESULTS

46. Eurotiox 32 Premix in meat and bone meal (Poultry origin)
Conclusions:
Oxidation is responsible for loss of nutrients, mainly energy
EUROTIOX 32 PREMIX is able to prevent nutrient losses due to oxidation.

47. Effect of EUROTIOX 32 PREMIX on energy values (kcal/kg) of MBM (ruminant origin)
Source: Trial AO 002: INDIA

48. Eurotiox 32 Premix versus competitor in feed (8% fat)
Competitor = ENDOX C