Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/10—Organic substances
  • A23K20/179—Colouring agents, e.g. pigmenting or dyeing agents
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/10—Organic substances
  • A23K20/158—Fatty acids; Fats; Products containing oils or fats
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K50/00—Feeding-stuffs specially adapted for particular animals
  • A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
  • A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
  • A23L33/15—Vitamins
  • A23L33/155—Vitamins A or D
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
  • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
  • A23L5/40—Colouring or decolouring of foods
  • A23L5/42—Addition of dyes or pigments, e.g. in combination with optical brighteners
  • A23L5/43—Addition of dyes or pigments, e.g. in combination with optical brighteners using naturally occurring organic dyes or pigments, their artificial duplicates or their derivatives
  • A23L5/44—Addition of dyes or pigments, e.g. in combination with optical brighteners using naturally occurring organic dyes or pigments, their artificial duplicates or their derivatives using carotenoids or xanthophylls

Definitions

• the invention relates generally to the absorption of carotenoids and, more specifically, to increasing the absorption and bioavailability of lutein and zeaxanthin in humans and poultry by the use of lysolecithin and lecithin.
• Carotenoids have long been used as important food coloring agents
• xanthophylls are added to poultry feeds so that upon ingestion, the xanthophylls are deposited in the skin and egg yolks Chickens specifically absorb
• lutein and its structural isomer zeaxanthin impart a yellow color to the chicken and its eggs that is commercially desirable
• Carotenoids have been hypothesized to reduce the risk of certain types of cancers in humans through their action as anti-oxidants that quench singlet oxygen and other oxidizing species thereby terminating free radical chain reactions and limiting cellular oxidation damage.
• lutein is among the most common and is known to exhibit strong anti- oxidant capabilities. The structure of lutein is:
• Lysolecithins (lysophosphatidylcholine, lysophosphatidylethanolamine, etc.) have been shown to have a variety of biological actions, all centered around modification of cell membrane permeability. Such effects include increased transfer of both cations and larger molecules across cell membranes in cultured cell lines (Duan, J. and M.P. Moffat (1991) "Protective effects of D.L-carnitine against arrhythmias induced by lysophosphatidylcholine or reperfusion", Eur. J.
• lecithins are phosphatidyl digylcerides, by which is meant that one of the three hydroxyl groups of glycerol (propane- 1, 2, 3-triol) is occupied by a phosphate group, which in turn is attached to a polar alkyl amine. The remaining two hydroxyls of glycerin are occupied by long-chain fatty acids.
• N cH 3 ) 3 is:
• lecithin as a phosphatidyl diglyceride, has both a polar, charged end and a highly hydrophobic one - the prime requirements of an emulsifier.
• Removal of the fatty acid in the center position by phospholipase A produces lysolecithins whose chemical properties differ noticeably from those of lecithins due to the greater hydrophilicity of its polar end. It has been found that the fatty acid remaining on lysolecithin is generally unsaturated (Dawson, et al. , ( 1990) "Fatty acid composition fo the neutral lipid and phospholipid fractions of mechanically deboned chicken meat", Poultry Science, 64: 1411-1419.
• the invention consists of a method and compound for increasing the abso ⁇ tion and bioavailabihty of carotenoids in humans and poultry by the use of lysolecithin and lecithin
• a formulation is made comprising the addition of a surfactant, including either lysolecithin or lecithin or both, to a carotenoid, including either lutein or zeaxanthin or a mixture of both
• the range of surfactant to carotenoid is between about 5% and about 30% by weight
• the formulation is fed as a food supplement and results in an increased abso ⁇ tion and bioavailabihty of the carotenoids 5
• An object of the present invention is to provide a food supplement including a carotenoid which increases the abso ⁇ tion of the carotenoid by an animal or human that is fed the supplement.
• Another object of the invention is to provide a method for increasing the abso ⁇ tion and bioavailabihty of carotenoids in food supplements.
• Still another object of the invention is to provide a method of increasing the abso ⁇ tion and bioavailabihty of lutein and zeaxanthin extracted from marigold petals.
• Figure 1 is a graphical representation of the results of NEPA analysis of egg yolks to determine pigment uptake in chickens fed a variety of diets including control diets and diets supplemented with feed additives of the present invention.
• Figure 2 is a graphical representation of the L* a* b* Color Space Figure in the Operation Manual of the Minolta Chroma Meter ⁇ .
• Figure 3 is a graphical representation of the results of reflectance colorimetric analysis of egg yolks from chickens fed lysolecithin-treated and untreated poultry feed.
• Figures 4 and 5 are a graphical representations of the results of NEPA analysis of egg yolks to determine pigment uptake in chickens fed a variety of diets including control diets and diets supplemented with feed additives of the present invention.
• Figure 6 is a graphical representation of the emulsifying properties of lysolecithin and measures the retardation of the phase separation over time. 6
• Carotenoids particularly the xanthophylls lutein and zeaxanthin, are important as colorants in animal feeds, particularly poultry feeds.
• Commercially available sources of the xanthophylls include the products of Kemin Industries, Inc., that are extracts of marigold (Tagetes erecta) and sold under the marks ORO GLO ® Liquid and FloraGLO ® Lutein (20% Liquid).
• ORO GLO ® Liquid has 7 grams of xanthophyll activity per pound (as determined by a method condensed from the Association of Official Analytical Chemists, paragraph 43.108, and available from Kemin Industries, Inc., part no. 02205).
• FloraGLO ® Lutein (20% Liquid) has 20% lutein and 0.86% zeaxanthin.
• the sources of xanthophylls have been used as additives to animal feeds to inco ⁇ orate the pigments into the body tissues and products of the animals.
• a method and formulation for increasing the level of abso ⁇ tion and bioavailabihty of the carotenoids has been developed.
• a formulation containing lutein and zeaxanthin is combined with lecithin and with lysolecithin at a concentration of between about 5% to about 30% by weight.
• the lutein and zeaxanthin source is mixed with the source of lecithin or lysolecithin and the mixture is added to feed for the animal.
• the treated feed is fed to animals for a predetermined length of time and the body tissues and animal products were assayed for xanthophyll content.
• the treated feed is found to increase the amount of the xanthophylls inco ⁇ orated by the animal by between about 2% and about 50% over control feed that included the same amounts of xanthophylls but which had not been combined with lecithin or lysolecithin.
• a quantity of a liquid source of xanthophylls was blended with a liquid source of lecithin or lysolecithin at high speed for an extended period of time to thoroughly mix the ingredients.
• the mixture was added to a low xanthophyll feed in 7 a mixer to assure uniform distribution.
• a control feed was formulated using the same low xanthophyll feed to which was added the same quantity of the liquid xanthophyll source also in a mixer to assure uniform distribution.
• the treated feed and the control feed were fed to separate groups of chickens for approximately one-month. Eggs were collected from both groups and analyzed using the NEPA test for pigment content.
• the pigment content of the eggs showed an increased pigment content in chickens fed the treated diet that correspond to chickens that had been fed an untreated diet including up to 50% higher concentrations of xanthophylls.
• the abso ⁇ tion and bioavailabihty of the xanthophylls was increased by up to 50%.
• Experiment 1 A feeding trial was conducted over a 28 consecutive day period. The trial used two hundred and twenty- four white Leghorn Hyline W36 cross birds, 29 weeks of age at the start of the study. The birds were housed in battery pens of eight or nine birds. All birds were fed a low-pigment diet for 1 month immediately prior to the start of the trial. Composition of the low xanthophyll diet used in this study is shown in Table 1 . Each treatment was fed to three replicate pens holding either eight or nine birds. The remaining birds received control low xanthophyll feed. Egg production was monitored throughout the test and remaining test feed for each treatment was weighed to determine feed consumption.
• the source of xanthophylls used in the trial was ORO GLO Liquid having a measured pigment content of 5.2 g/lb xanthophyll activity.
• the source of lysolecithin used was Lysoprin-brand lysolecithin purchased from Lovesgrove Research (Aberystwyth, Wales), product no. 10544.
• Target application rates for ORO GLO and Lysoprin are shown in Table 2. For the treatments with Lysoprin, 0.78, 1.17 and 1.56 lb of ORO GLOliquid were added to 0.23 lb of Lysoprin. The mixture was blended at high speed in a Waring blender for 15 minutes.
• Each mixture was poured, during mixing, into a ribbon mixer containing 195 lbs of low xanthophyll feed which had 10 been previously treated with 150 ppm TERMOX ® dry (from Antitox, Buford, Georgia) and 1,000 ppm Myco CURB ® liquid (from Kemin Industries, Inc.).
• the diets thus produced contained the equivalent of 40, 60 and 80 grams of xanthophyll activity per ton of feed, respectively.
• Control feed was prepared by addition of 0.78, 1.17 and 1.56 lb of ORO GLO liquid directly to low xanthophyll feed without addition of Lysoprin. Samples of each feed were taken and analyzed for pigment content using a method based on AOAC, Paragraph 43.018, without saponification and column chromatography.
• treatment levels will be referred to by the target level of total xanthophyll activity in grams of xanthophyll per ton of feed as desc ⁇ bed in the Methods and Mate ⁇ als section and Table 2
• Egg collection data feed/bird/day and average egg weight
• NEPA National Egg Products Association which originally developed it In modified form, it has been accepted as a standard method by the AOAC (Journal of the
• colorimeter data is obtained and reported as the values L*, a* and b* which correspond to the position of the reflected color in a three-dimensional system.
• the L* value is a measure of brightness (luminosity) and a* and b* are chromaticity coordinates along green-red and yellow- blue axes, respectively.
• the coordinate system is shown graphically as Figure 2. While actual color changes can most accurately be presented on a three-dimensional graph, it is common practice to report the individual L*, a* and b* values.
• the L* a* b* system is designed to mimic human color perception.
• the relatively small changes in L*, a* and b* upon addition of pigment correlates well 15 with the known difficulty in perceiving color changes at the pigment levels used m this study Conside ⁇ ng the lack of hnea ⁇ ty of response within the CIE t ⁇ angle to yolk color changes, the high degree of lmea ⁇ ty in the a* value response for untreated feed ( Figure 3) with increasing pigment levels observed in this study should be noted
• the correlation coefficient (R 2 ) from untreated feed is 0 999, it is 0 973 from lysolecithm-treated feed
• the ORO GLO liquid used for the trial had a pigment content of 5.32 g/lb.
• Target application rates for ORO GLO, Lysoprin brand lysolecithin and bulk soy lecithin are shown in Table 6.
• For the treatments with Lysoprin and lecithin 55, 1 10 or 165 grams of either Lysoprin or lecithin were blended with 550 grams of ORO GLO liquid. The mixture was blended at high speed in a Waring blender for 7.5 minutes and at low speed for 5 minutes. The ORO GLO used in the control treatment was not blended.
• Treatment was affected by pouring each of the above pigment mixtures onto feed in a ribbon mixer containing 210 lbs of low xanthophyll feed which had been previously treated with 150 ppm TERMOX dry and 1,000 ppm MYCO CURB liquid.
• Control feed was prepared by addition of appropriate amounts of ORO GLO liquid, as outlined in Table 2, directly to low xanthophyll feed without addition of Lysoprin or lecithin.
• Each treated feed was run through a hammer mill and mill screen to aid in pigment distribution, then weighed in lots of 35 lbs into poly- lined Kraft bags. Samples of each feed were taken and analyzed for pigment content using a method based on AOAC, Paragraph 43.018, without saponification and column chromatography.
• Prin Lysoprin brand lysolecithin.
• BCE ⁇ Carotene Equivalents a-d Ent ⁇ es in column with no common supersc ⁇ pts differ significantly (P ⁇ 0 05)
• a feeding t ⁇ al was done for 28 days Two hundred and twenty-four white Leghorn Hyline W36 cross birds were used, 47 weeks of age at the start of the study 20
• House temperatures ranged from 58° to 72°
• the ORO GLO liquid used for the trial had a pigment content of 5.4 g/lb.
• Target application rates for ORO GLO, LYSOFORTE and Lysoprin lysolecithin supplements are shown in Table 9. Since LYSOFORTE is a 10% suspension of Lysoprin on an inert carrier, application rates for LYSOFORTE were set at 10 times those for Lysoprin. Thus, a 2.2 lb/ton treatment of LYSOFORTE is designed to parallel a 0.22 lb/ton application of Lysoprin.
• Lysoprin For the treatments with Lysoprin, 10.5, 42 and 105 grams of Lysoprin were blended with 352 grams of ORO GLO liquid. The mixture was blended at high speed in a Waring blender for 15 minutes. Treatment was affected by pouring each of the above pigment mixtures onto feed in a ribbon mixer containing 210 lbs of low xanthophyll feed which had been previously treated with 150 ppm TERMOX dry and 1 ,000 ppm MYCO CURB liquid. LYSOFORTE treatments were prepared by adding 105 and 420 grams of LYSOFORTE ( 10% Lysoprin) to the mixer after adding pigment.
• Control feed was prepared by addition of appropriate amounts of ORO GLO liquid, as outlined in Table 2, directly to low xanthophyll feed without addition of Lysoprin or LYSOFORTE. Samples of each feed were taken and analyzed for pigment content using a method based on AOAC, Paragraph 43.018, without saponification and column chromatography. 21
• Inco ⁇ oration of Lysoprin lysolecithin into pigment-supplemented poultry diets for active layers yields statistically valid increases in the pigment content of egg yolks. Based on the above data, a pigment level of 40 g ORO GLO liquid/ton of feed, with addition of lysolecithin at the application rates used in this study, will show yolk color performance between that of 40 g/ton and 60 g/ton untreated feeds. Conversely, addition of LYSOFORTE dry lysolecithin supplement did not improve pigment inco ⁇ oration.
• the experimental diets were prepared in the following manner. Five samples of 210 lbs of the low-xanthophyll diet of Table 1 were supplemented with one of the following: 2.2 lbs/ton of Lysoprin , an alternative commercial product (Blendmax 322D liquid), 11 lbs/ton LYSOFORTE, and an untreated control. In addition each sample was treated with 8 lbs/ton ORO GLO® brand liquid and 2 lbs/ton MYCO CURB® brand liquid. The liquid lysolecithin products were mixed with the ORO GLO liquid in a Waring Blender for one minute before treating the feed.
• the lysolecithin and/or pigment products were added to the feed at the indicated levels by slowly pouring the additive onto feed during mixing in a ribbon mixer. After running through a hammer mill with a mill screen, 35 lbs of each treatment was weighed into poly-lined Kraft bags. 25
• the quality assurance test were carried out in the following manner. 1.6 g of liquid Lysoprin and 8 g of dry LYSOFORTE are added to separate 100 ml samples of canola oil. These solutions were then mixed thoroughly, after which the dry LYSOFORTE was filtered to remove all carrier material. At the same time a 15% NaCl solution was prepared. Fifty ml of the lysolecithin solutions and 50 ml of the 15% NaCl solution were added to a 150 ml beaker. The materials were mixed with a 10,000 rpm bio-homogenizer for 15 seconds. The emulsion was immediately transferred to a 100 ml graduated cylinder. The volume of the lower phase (the free 15% NaCl solution) of the emulsion was observed at time 0 and then every five minutes over a 60 minute period. A blank of canola oil and the 15% NaCl was also run for comparison.
• the yolk fan scores were conducted first. Ten grams of the composite egg yolk material from each treatment was placed in a plastic 60 x 15 mm Falcon 1007 petri dish. Six individuals in the research facility were then asked to determine the color fan score of each yolk composite. The fan scores were then analyzed at the 95% confidence level using Duncan's multiple comparison procedure (Statgraphics Plus, Manugistics, Inc., 1995).
• the intensity (L), red (a), and yellow (b) pigmentation was measured using a CR-300 Minolta Colorimeter. Again, 10 g of the composites egg yolk from each treatment was placed in a plastic 60 x 15 mm Falcon 1007 petri dish. To measure the L*a*b of each treatment the sample was placed on the top of the measuring head with 26 an open o ⁇ fice for color head Measurements of L*a*b were replicated twice for each sample The L, a and b scores were then analyzed at the 95% confidence level using Duncan's multiple comparison procedure
• Lysoprin (lot 200795) 80.61 80.48 80.54 e 15.6
• Lysoprin (lot 250795) 74.06 73.12 73.59° 5.6
• the results of this trial also indicate that there is a positive, although limited, correlation between the results of the quick assay and the results of the layer trial.
• the three liquid Lysoprin lots that performed well in the layer trial also performed well in the quick assay while the dry LYSOFORTE product failed in both, the quick assay as well as the layer trial.
• the quick assay failed to predict the order of performance that was seen in the layer trial (e.g., Blendmax 322D performed best in the quick assay but was inferior in the trial).
• Blendmax 322D performed best in the quick assay but was inferior in the trial.
• the emulsifying properties of the lysolecithin products are consistently seen only in canola oil, not however in corn oil or mineral oil.
• the Blendmax 322D product showed a good effect also in mineral oil. 29 Experiment 5
• Temperature in the house during the trial ranged from 20° C to 32° C.
• MYCO CURB® brand liquid was also added to each treatment at 0.916 kg/ton. Mixing of the feed samples was done in a ribbon mixer at low speed. Each treatment was weighed into poly-lined Kraft bags, after running through a hammer 30 mill with a mill screen Samples were taken and analyzed for pigment content using QC method QPM-10 without saponification or chromatography
• Pigment conversion (%) 100% x (BCE) x (yolk weight) x feggs/da /hen) (Pigment) x (feed/hen/day)
• Tables 16 displays the results of the quantitative determination of the pigments and pigment conversion in the egg yolks Addition of ORO GLO to the low pigment
• the diet resulted m a 35 0% better pigment conversion as compared to the low pigment diet
• lecithin or Lysop ⁇ n up to a final concentration of 0 916 kg/ton feed were added with the ORO GLO dry to the low fat or the high fat diet
• lecithin showed the greatest increase in pigmentation and pigment conversion (22.2% over the ORO GLO control) for the low fat diet
• Lysoprin resulted in the best increase for the high fat diet (23.3% over the ORO GLO/high fat control).
• surfactant treatments of the low fat diet were not statistically different among each other.
• the egg productivity of the laying hens was also monitored for the duration of the trial. The results are displayed in Table 17. No significant differences in egg productivity were seen for the various treatments.
• a high fat diet was observed to lead to higher pigment utilization and pigment conversion as compared to a low or no fat diet. It further appears that while lecithin performs better in a low fat diet, Lysoprin performs better in a high fat diet as measured by pigment conversion.
• the first observation can be explained reasonably by the fact that uptake by the bird and deposition of the hydrophobic carotenoid molecules in the egg yolk is more effective when dispersed in fat in the animal's gut.
• the second observation seems to reflect a more specific effect with much higher efficacy for the lecithin and Lysoprin as compared to the soybean oil/poultry fat blend.
• Lysoprin can display full efficacy as an emulsifying agent only in a diet supplemented with a certain amount of fat.

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• 1999
  • 1999-03-17 EP EP99912665A patent/EP1063898A4/de not_active Withdrawn
  • 1999-03-17 WO PCT/US1999/005925 patent/WO1999047001A1/en not_active Application Discontinuation
  • 1999-03-17 MX MXPA00009116A patent/MXPA00009116A/es not_active Application Discontinuation
  • 1999-03-17 AU AU30988/99A patent/AU745973B2/en not_active Ceased
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