MXPA01001228A - Method for the conversion of xanthophylls in plant material - Google Patents

Abstract

This invention relates to the in situ process for converting non free form xanthophylls to free xanthophylls in the biological material of the plant. The method more particularly relates to a method that would liberate xanthophylls by transesterification of acyl-xanthophylls in plant materials.

Description

METHOD OF COISTVERSION OF XANTOPHILES IN MATERIAL FOR PLANTS DESCRIPTION OF THE INVENTION This invention refers to the in situ process to sonvert forms of non-free xanthophylls to free xanthophylls in the biological material of the plant. The method relates more particularly to a method that could liberate xanthophylls by transesification of acyl xanthophylls in plant materials. Carotenoids are a group of red and yellow pigments that are found in plants and fruits. Sarotenoids include carotenoids and hydroxylated carotenoids designated as xanthophylls. Xanthophylls include lutein, zeaxanthin, capsorubin, oapsanthin, astaxanthin, and santaxanthin. The animal feed industry and the food industry and the pharmaceutical industry have all had a strong interest in xanthophylls. The sarne industry receives a benefit by adding xanthophylls to increase the color of the yolk of the eggs. Pharmaceutical companies have found that xanthophylls are useful in certain tumor treatments and as an antioxidant. The food industry has found that consumers are looking for food dyes that occur naturally. Annatto, saffron and paprika are some natural carotenoids that have traditionally been used for food coloring. Intense red and yellow coloration and xanthophylls that occur naturally in edible plants such as vegetables and fruits such as biocolli, green beans and Brussels pea and suns, cabbage, curly sun, spinach, iwi and honeydew have attracted many uses including a pigmentation additive for animal feed. In certain plants xanthophylls are in the non-esterified free form. However, the large amounts of sorophylls in vegetables have difisil concentration or extraction of xanthophylls. A number of xanthophylls is also present in yellow solor fruits and vegetables such as mango, peaches, plums, papaya, papaya and oranges. These are less corollas, but often xanthophylls exist in esterified form with fatty acids such as myristic, lauric, and palmitic acids. Unfortunately, to be metabolized into a feed additive, the xanthophyll ester must undergo conversion to free xanthophylls. The free xanthophyll can then be metabolized by the body. Various plant materials contain xanthophylls; The desired xantofile's will operate the selection of the material used. For example, it is well known to use the petals of the calendula flower. Tagetes erecta for the lutein extraction of xanthophylls. Calendulas are easily cultivated and have been used as a source of pigment for meats. Lutein occurs in the marigold flower diacylated with pamymitic and myristic acids, in large fatty acid esters typically as diesters in the syromoplasts. The animal feed industry has taken two different approaches to provide xanthophylls in animal feed, particularly lutein to sarne feed. The industry has used dried marigold food as a food additive thus providing lutein in the less used acylated form. This form of xanthophylls requires more consumption of calendula food to achieve the desired pigment. Alternatively, the industry has used a number of processes, starting with the extraction of the xanthophylls from the plant material and the oleoresin formation. The industry subsequently adds and prospers the oleoresins to sonvert the xanthophils from the asylated form to the free form by a number of different processes including transesterification for some oleoresin processing of paprika, although for lutein the process is mainly by saponifissation . The oleoresin is converted, requires less consumption by the animal to achieve the desired xanthophyll. Nevertheless, the oleoresins' formation and the prosesamiento of the same ones by saponification is both the consumption and aid in the elaboration costs for the alimentary product. La = aponifisasión is the conversion of the fatty acid into a soap by treating it with an alkali. The saponification number is the number of milligrams of potassium hydroxide required to saponify one gram of the ester. After saponification, the industry has often used solvents to crystallize lutein from oleoresin. This has made xanthophylls more pure and available for the body that consumes lutein, but has added tierrypo and labor to the process of supplying xanthophylls to the food mixture. Some of the following patents indicate the processes for recovering various compounds such as xanthophylls from oleoresins. U.S. Patent 5,602,286 describes a process for the recovery of xanthophylls from corn gluten. The patent has The steps of: adding ethanol as an extra stage, filtering, then peeling to form crude xanthophyll and then using ethanol, KOH as the saponification step, then washing and filtering, then purifying the refined xanthophylls. Three Japanese references also show the use of similar oleoresin extractions. No. 82,133,160, from Japan 182, shows a production of red pepper pigment using an oleoresin of red pepper, in either water or alcohol mixtures, treated with KOH, NaOH, CaCO3, and then treated with acids such as HCl, H2r S04, H3P04 HOAc, lactic acids and citrus. The pigment solutions are removed with organic solvents such as MeOH, EtOH PrOH and acetone. Patent No. 81,180,663 (1982), show the paprika, food coloring agents that are extracted as an oleoresin. The oleoresin was heated with basic alkali metal compounds such as KOH, NaOH, K2C02, Na2Co3, and sodium allooholate, and one or more hydroxides and carbonates or alkaline earth metals such as Ca (OH) 2 are mixed. The precipitates were extracted with organic solvents and produced an odorless oleoresin pigment. Patent No. 1.3,173,164 shows paprika pigments that can be prepared by treating paprika oleoresin with alkali at temperatures below 50 ° C in the presence of halogen, sulfate, bicarbonate, carbonate, phosphate, and aliphatic carboxy ions, and then treated with an organic solvent and finally extracted with acetone. U.S. Patent 5,382,714 discloses a method for producing substantially pure lutein. In this patent the starting material was calendula petals. The process of saponification of the petals is briefly described in column 5, example 1. The petals of the flower were tested by herbicides and pesticides and then the xanthophylls containing the material were subjected to saponification with aqueous potassium hydroxide. This was achieved by continuous mixing by heat (65-70 degrees C) of potassium hydroxide at 45% feed grade. This achieved the 98% conversion of letein in a form that was free of fatty acids and is presented as a yellow oil. This material could then be used as a food or food additive. The present invention provides a method of in situ conversion of xanthophylls into the free form for the liberation of xanthophylls by transesterification. Thus, the present invention avoids the need for the formation of oleoresin. This oleoresin conversion requires extractions of organic solvent from the material from the plant material. Hexane is used frequently. The present invention provides free-form xanthophylls in situ by transesterifissation of the material in situ in this manner eliminating the need for an oleoresin or saponification of the material. The present invention allows the calendula food to be subjected to transesterification and then used without the extraction of xanthophylls from food. It is evident that there is a need to convert acylated xanthophylls in plant material to the free form. The object of this invention is to fulfill that need. Another object of this invention is to provide a marigold food that has a high content of free lutein. Yet another object of this invention is to provide a marigold food as a feed additive that provides additional pigment to the eggs when compared to the same marigold food as a feed additive that has not undergone the treatment of the present invention. Another additional object of this invention is to provide a plant material containing sustansialmen; e a higher percentage of the free form of xanthophylls, then contained in the original plant material before release. Broadly, then the present invention includes an improved plant material made from a natural plant material. This plant material contains at least part of the xanthophylls in a non-free form, comprising plant materials that contain less xanthophylls in a non-free form, in situ, and more free-form xanthophylls in which the free form of xanthophylls in situ In the improved plant material it has increased beyond the amount in the natural plant material. The invention may have a number of different plant materials including flowers and flower petals.

A flower of particular utility is the Tagetes erecta. This type of flower is useful if the desired xanthophylls are lutein.

The volume of xanthophylls in marigolds exists in nature as non-free xanthophylls, such as fatty acid esters. The present invention is adapted to convert non-free xanthophylls into the free form of xanthophylls. When the calendula flowers are used, this conversion by transesterifisation produced a non-acylated lutein. The present invention can improve the plant material so that it contains at least 5% more free-form xanthophylls than the natural plant material in situ. Another embodiment of the present invention is a composition d = improved animal feed comprising vitamins and minerals, together with a source of carbohydrates selected from the group consisting of soy, casahuate, corn, alfalfa, wheat, barley and, improved plant material containing xanthophylls in situ more freely, then the natural in situ amount of xanthophylls in the plant material that the improved plant material was formed from, wherein the xanthids are more bioavailable for an animal feed to feed the animal. This food may contain improved plant material that is flowers. If the desired xanthophyll is lutein, then the flowers are marigolds. If the desired xanthophyll is sapsanthin, then the paprika can be used. This animal feed is designed for the nutritional requirements of poultry, where animal products such as eggs and meat may have xanthophyll which acts as a pigment. It is particularly believed that the present invention is useful for chickens. Certain xanthophylls could also be useful in other animal feeds to color meat or other animal by-products. An increase for poultry is preferably characterized by having feed for animals which makes evident the thioavailability of the free form xanthophylls having increased pigmentation from the consumption of free form xanthophylls, especially when the feed xanthophylls are lutein. The present invention encompasses a product and also the method of forming free form xanthophylls. Thus, broadly, the present invention covers a method of the improved plant material made from a natural plant material containing at least some non-free xanthophylls by the following steps: treating the natural plant material with a solvent in situ; add a base capable of transesterification of xanthophylls in a non-free form to free form xanthophylls; neutralizing the reaction where the improved plant material that has more free-form xanthophylls is formed, then the natural plant material. The method can also include the step of drying the improved plant materials to remove any solvent. The method of the present invention includes using p Lanta material such as flowers. If desired, the product is lutein, then flowers such as Tagetes erecta can be used. The method uses a solvent in the reaction. The solvent can be widely a alsohol and more preferred an alsanol or alkenol. If a alcohol is also used, the alcohol preferably has one to one carbon atom. The alcohol is selected from the group consisting of ethanol, ethanol, isopropyl alcohol, butanol and the like. The method of the present invention uses a base.

The base can be selected from the group consisting of potassium hydroxide, potassium sorbate, NaOMe, animal liver lipase, yeast lipase, NaOEt, KOMe, KOEt, Na2C03, K2C03 and the like. The method also includes the stage of stopping reaction by neutralizing the reassessment of Lewis' acid. The preferred Lewis agarido is a rough phosphoroid. However, the Lewis acid may be sequestered from the group consisting of HCL NH4CL, acid sulfur, ALCL and the like. The method does not require an extrastor such as petroleum ether or hexane or a number of other extraneous salts to remove xanthophyll from the plant material.

The method can have a plant material such as flowers, roots or fruits, but preferably without chlorophyll in the product. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a calendula food chromatogram. Figure 2 is a paprika chromatogram. Figure 3 shows the chromatogram of conversion to the free form of capsanthin from treated paprika. Figure 4 shows the chromatogram of conversion to the free form of lutein from the treated marigold food. Figure 5 shows the Roche fan scale for days for the color of the egg yolk of the present invention at two mg / kg levels and the comersial product. Figure 6 shows the mg / kg of xanthophylls in the egg yolks during the days of the three treatments. This invention refers to the in situ process to convert non-free xanthophylls to free xanthophylls in the biological material of the plant. The most particular method relates to a method that could liberate xanthophylls by transesterification of acyl xanthophylls into plant materials. And then the improved plant material that has xanthophyll more freely, the original plant material from which it is made. Thus the present invention provides a method for releasing xanthophylls in situ in plant or vegetable material or fruit. A xanthophyll in the free form such as lutein, zeaxanthin or capsanthin is formed from the xanthophyll diester in the plant material. The plant material should have relatively concentrated amounts of the desired xanthophyll in the non-free form that is usual and preferably in the form of a fatty acid ester. Calendulas are: an excellent source of lutein in the form of diesters. Also currently known in the wild are the white berry fruit (Lycium barbarum) an excellent source of xeaxanthin diesters, and the pepper plant (Capsicum annuum) an excellent source of capsanthin as diesters. Other plants and fruits and vegetables have high concentrations of desired xanthophylls can also be used. The term "xanthophyll" in a non-free form refers to a xanthophyll which is in a form that can be transesterifylated to the free form of xanthophyll. The term "free form xanthophyll" refers to a xanthophyll which is not in an esterified form. The term bioavailability refers to the extent to which xanthophyll is available to the body of the body that consumes it.

The term "base" refers to potassium hydroxide, potassium sorbate, NaOMe, animal liver lipase, NaOEt, KOMe, KOEt, Na2C03, K2C03, and such non-nucleophilic / non-nucleophilic deprotonation material, sual does not provide deprotonation kinetics in the alpha position of the carbonyls and highly conjugated double bond systems, for example, LDA - or BuLi could be excluded from the base definition because these chemicals cause the deprotonation in the alpha position of the carbonyls and the systems of double bond highly conjugated. The term Base Ratio to plant material (Food) by Weight - refers to the amount of base used in the reaction compared to the weight of the plant material. This amount is determined by the pH of the reaction and the desired withdrawal times. The preferred pH is between 11-14: at a lower pH that can be used, but which increases the reaction time. The term "solvent" refers to a chemical in which the transesterification of xanthophyll can be carried out, the chemical preferably having a hydroxyl group. The solvent is preferably a alsohol with 1-4 carbons. More preferably, the solvent is selected to have a boiling point which allows the 7th transesterification reaction temperature to be maintained at 75-85 degrees C, such as MeOH.

Plant material refers to plants that contain xanthophylls in the non-free form of xanthophyll. The contemplated plant sources contain xanthophylls in the esterified form as a long chain of mono- or di-C? 2-Ci8 of fatty acids such as lauric acids, mirísticos, oleicos, linolenisos, and palmitic. Sallendulas are an excellent source of lutein in the form of diesters now known in nature, the white berry fruit (Lycium barbarum) is an excellent source of zeaxanthin diesters, and the paprika plant (Capsicum annuum) has capsanthin in the form of diesters. Other plants and fruits and vegetables having high concentrations of desired xanthophylls may be employed. It is well known in the art to extract carotenoids from plants. It is equally well known in the art that the food of calendula and certain other plant materials can be animal feed as a feed additive to cause the pigmentation of the product, animals such as eggs or meat. The calendula meal has been supplied to the chicken egg pigment for a long period of time. Lutein has been extracted from marigold for the properties of pigmentation. What the previous teasin has taught and suggested is that there are two ways to supply lutein. One is the plant material that naturally wears with lutein in the non-free form. Alternatively, extract the lutein by forming an oleoresin and saponifying the oleoresin and supplying the saponified material having the iodine form of xanthophyll. The present invention provides a new and better alternative. The present invention is a method of delivering an increased amount of free formed xanthophylls, in situ, into the plant material, without having to form the oleoresin. The prior art teaches that organic extractors have been used to extract carotenoids from the plant material. Such extractors include hexane, acetone, petroleum ether, methanol, ethyl acetate, diethyl, heptanes, chloroform, and tetrahydrofuran. These extractors result in what is called an oleoresin which contains diés. The prior art teaches then the use of a saponification reaction that cleaves the fatty acids from diesters of xanthophyll. There are a number of known methods for saponification. These produce free-form xanthophylls together with fatty acid soaps. The soaps are made with alkaline solutions such as potassium hydroxide and sodium hydroxide in aqueous solutions. Although similar chemistries can be used in the saponifisation process, the present invention does not require the step of extractions or the saponification step to provide an improved plant material. The release of xanthophylls in plant material in situ proceeds by the following chemical equation: Clearly, diacyl xanthophylls are converted by transestei stasis in free xanthophylls in situ (in the plant material srudo). This process avoids the cost and potentially hazardous processes and chemicals associated with the production of an oleoresin. Additionally, the plant material can act as the carrier material increasing the efesia of the process. Thus, the steps of the present method include colosating the mat, which is the non-free form of xanthophylls in the solvent and base. The base ratio to p.anta (Food) material by weight is the amount of base used in the reaction compared to the weight of the plant material. This amount is determined by the pH of the reaction and the times of; desired reacción. The preferred pH is between 11-14: the lower pH invests the reaction time. During approximately every 32 grams of xanthophyll activity in the pure plant material about 770 g of base such as KOH was dissolved in 11 liters of the solvent. The solution containing the plant material was maintained at a pH of about 13, between 11-14 with acceptable levels being 13 being the preferred level. The reaction solution was stirred until full conversion was observed. The operating time was approximately 10 hours at 69 ° C and was verified by HPLC. The reaction solution was neutralized to a pH of 7, which is highly phosphoric. Any number of neutralisers and agents could be employed. But phosphoric acid is preferred. The solvent, preferably an alcohol and more preferably MeOH was removed with 16 hours of distillation at 69 ° C. The distillation temperature can be as high or as low as in the previous to the boiling point of MeOH and the distillation is carried out in a reasonably reasonable time. Other carriers may be added to the plant residue material such as almond food, silicates and the like. The residue was dried after distillation of the solvent, either at room temperature to an ATM or by vacuum drying in an oven at < 50 ° C temperature. The method of scouring is a matter of drying time and is not critical. The plant residue material that now has free-form xanthophylls in situ herein is mixed and a fine powder with a xanthophyll activity of 10-14 g / kg is achieved. The final product is preferably stored at room temperature under nitrogen. The following bases may also be useful in the present invention. Their selection can be based on the economy of the process, the speed of the process and the acceptability of the trace quantities of the components. The present invention having a free-form xanthophyll plant material can be marigold food that has been converted. The converted marigold food was fed to the sorral birds along with a control (the same marigold food from the same batch of plant materials), the same type and age of the egg-laying hens. The data indicate that the use of xanthophylls in the converted food was increased. In other words, there was more pigmentation of the egg yolks when the same amount of the converted plant material was consumed compared to the non-converted plant material. Thus, the process will provide xanthophyll in a greater bioavailable form from the animal consuming the product. Generally, this test was run with marigold food which was processed by the stages listed below to provide free-form xanthophylls in situ.

Experiment 1 Lutein release by transesterification Step 1. Marigold meal (2 kg with a total xanthophyll activity of 16 g / kg) in MeOH / KOH (11 L / 770 g, pH = L3) was stirred until the complete conversion was observed (approximately 10 hours) at 69 ° C. The reaction production was checked by HPLC. See Figure 1 and Figure 3. Step 2. The reaction mixture was neutralized with phosphoric acid (pH = 7) and the MeOH was distilled off at 69 ° C (distillation time = about 16 hours). Step 3. The residue was then dried in a well-ventilated room at room temperature and at 1 ATM or by vacuum drying in an oven (temperature <50 ° C). Step 4. After mixing and intermixing, a fine polymer with a total xanthophyll activity of 10 to 14 kg / kg was obtained (depending on the duration of the reaction and the drying process). The final product is stored at room temperature under nitrogen. The process described above may use a number of different bases including, but not limited to potassium hydroxide, potassium sorbate, NaOMe, animal liver lipase, NaOEt, KOMe, KOEt, Na2C03, and K2C03 and other deprotonation material non-nucleophilic and kinetically without force which does not cause deprotonation in the alpha position of carbonyls and highly conjugated double bond systems. The preferred base is KOH because of its availability at a non-expensive price and it is effective in avoiding deprotonation emissions. Holiness for the use of other bases is easily determined. As will be observed, the process described in the following experiments does not involve the use of an aqueous solution. Instead, the solvent is an alcohol solvent that, besides, is the process itself, which can be removed by distillation. MeOH is preferred as a solvent, but other types of alcohol, such as: isopropyl alcohol, ethanol and butanol and the like can be employed without undue experimentation by the person of ordinary skill in the art. The solvent is clearly a chemical in which the transesterification of xanthophyll can be carried out. The solvent preferably has a hydroxyl group and is not an aqueous solution. The neutralizing agent is preferably phosphoric acid. But as a neutralizing acid, other Lewis acids may be used, such as: HCL, Sulfuric acid, A1C13, NH4CL, acetic acid and the like. The plant material refers to plants that contain xanthophyll in xanthophyll in a non-free manner. The contemplated plant sources contain xanthophylls in the esterified form as a long chain of mono- or di-C? 2-C_8 fatty acids such as lauric acids, myristyses, oleisos, linolens and palmitic. Xanthophylls are found in a number of different plant materials. The calendula has lutein and the paprika has capsanthin. This process works to release a number of xanthophylls in a non-free manner in plant materials. The next experiment is the reactive procedure for the release of capsanthin in paprika meal. Experiment 2 Release by means of the transesterification of paprika to form free capsanthin The material of paprika plant having a total xanthophyll activity determined per kg was placed in MeOH (11 L, without ethoxyquin) and stirred for 8 hours at 69 ° C and for 12 hours at room temperature with KOH (770 g, pH = 13). Reaction production was verified by HPLC. The reaction mixture was neutralized with phosphoric acid (pH = 7). The MeOH (6 L) was distilled off at 69 ° C (distillation time = 16 hours at 69 ° C with a 48 hour pause at room temperature). The residue was then dried on standing at room temperature at 1 atmosphere, and was followed by vacuum drying in an oven (50 ° C, 100 Torr) for 2 hours. After mixing, a fine powder with a total xanthophyll activity was obtained which is determined per kg (depending on the duration of the drying process and the eficasia of mixing). Analysis: It can be done with the following parameters: - Verify the transesterifisation: Chrompack chromsep 100 * 46 mm (L * ID) 150! .8 misrospheres C18, number sag. 28076 flow 1 ml / min. eluent: CH2C12 / CH3CN 30/70 exp.oration at 450 nm The total xanthophyll activity was measured by means of a spectrophotometer. Experiment 3 Comparison of calendula food chromatography with transesterifed marigold food to evaluate free lutein The transesterification process described above in experiment one was used in one of the marigold food samples, the other sample was not treated by the esterification process. The two materials of the plant material that have been treated and the untreated material were analyzed and the results are shown in Figures 1 and 3. Figure 1 shows the untreated marigold food in and Figure 3 shows the marigold food treated. Figure 3 was subjected to the following conditions: Column Pressure (PSI): 1866 Column Temperature (C): N / A Noise (microAÜ): 3e + 001 Trend (microAU / min): 3e + 001. the untreated marigold meal had the following parameters and condiions: Column Pressure (PSI): 2391 Column Temperature (C): N / A Noise (microAU): 4e + 001 Trend (icroAU / min): le + 001 La graph in Figure 1 shows that the lutein of calendula somes has a higher ester activity in the range of 28-33 and a lutein level in the range of 7-8. The transesterified marigold food shows that the lutein peak is still high and the esters in the area of 31-33 no longer exist. The following data was collected from the calendula meal and Figure 1.

Component T? Area Height Area% Tino of DÍCO UnidentOOOl 2.513 11 10 194 O.05 Resolved Unident0002 4.220 4408 1331 0.19 Cast UnidentOOOS 4.365 2763 670 0.12 Cast Unident0004 4.484 17210 2068 0.76 Fade CAPSANTHIN 6.602 8032 758 0.35 Fade Unident0007 6.805 6003 891 0.26 Cast UnidenlOOOß 7.054 794979 90124 34.96 Cast LUTEIN K1 7.862 85277 3973 3.75 Fade ZEAXANTHIN 8.110 123244 7773 5.42 Fade RUBD ANTUC 8.797 15407 1288 0.68 Cast CANTHAXANTMN 9.900 0 0 0 NF CITRAXANTHGN 1 1.470 3141 223 0.14 Solved1 B CRYPTOXANTHIN 12,626 8967 478 0.39 Resolved LYCOPENE 17.959 27086 689 1.19 Cast B CAROTENE 19,839 41511 2125 1.83 Cast Unident00I7 20,163 17368 1290 0.76 Fade UnidentOOlß 21.777 12620 719 0.55 Fade U? Üdent001 22.138 6278 502 0.2S Fault Unident0020 28.596 16349 1191 0.72 Resolved Unident0021 30.268 92474 6473 4.07 Fade Unident0022 31.082 9061 802 0.40 Fade U? Üdent0023 31.445 2991 321 0.13 Fade! Unident0024 31.816 228688 16904 10.06 Fade Ur_.den.J025 32.225 13487 1254 0.59 Fade Unident0026 32.604 45440 2610 2.00 Fade U? Üdent0027 32.847 10055 1418 0.44 Fade Unidenl0028 33.184 408509 31639 17.96 Fade Unident0029 33.564 31417 2652 1.38 Fade Unident0030 33.940 79024 4915 3.48 Fade U? üdent0031 34,533 161146 8099 7.09 Fundid Tdíáls 2274045 193374 100.00 Comoonente RT (min) Area Altura Area%. TÍDO de Pico Unident0002 4,328 57885 4754 0.54 Resolved: CAPSANTHIN 6,454 17237 901 0.16 Fade Unident0004 7.130 94538 8757 0.88 Cast LUTEIN K1 7.459 7571877 942410 70.17 Fade ZEAXANT? IN 7.959 1247667 54701 11.56 Cast RUBIXANTHIN 8.634 1533918 92877 14.21 Cast CANTHAXANTUIN 9,311 72773 5608 0.67 Cast CITRAXANTHIM 11.976 51048 1889 0.47 Cast B CRYPTOXANGHIN 12.475 9046 880 0.08 Cast UnidentOOH 13.231 87645 3194 0.81 Fade LYCOPEEN 16,350 0 0 0 NF B CAROTEEN 19,830 28395 1187 0.26 Resolved Unident0014 20,776 5077 454 0.05 Resolved Un¡dßnt0015 31,145 2061 238 0.02 Resolved Unident0016 32,573 3773 477 0.03 Resolved Unident0017 33.801 8411 754 0.08 Resolved Totals 10791351 1119081 100.00 Experiment 4 Comparison of paprika chromatography with transesti fi ed paprika to evaluate 1 free capsule The transesterification process described above in experiment 3 was used in one of the paprika samples, the other sample was not treated by the transesterification process. The two materials of the plant material: 1, which has been treated and 2, the untreated material were analyzed and the results were shown in Figure 2 and. Figure 2 shows the untreated paprika and Figure 4 shows the paprika treated. Figure 4 was subjected to the following condi tions and gave the following data.

Transesterified paprika Acquisition Trunk Column Pressure (PSI) 1799 N / A Noise (microAU) 5e + O01 Component TA (min) Area AKHGÍI Area% Peak type Un_dent0002 3,350 5402 305 0.46 Cast U? Udent0003 4.212 75241 6091 6.47 Cast Unidem0004 5.939 167992 10409 14.44 Cast Unident? 0O5 6.189 293468 17946 25.22 Cast CAPSANTHIN 6.455 89525 8942 7.69 Fundidc Unident0007 6.640 93922 9488 8.07 Cast Unidenl0008 7.028 163104 13380 14.02 Cast LUTE1N K.1 7.936 4794 380 0.41 Fade ^ EAXANTHIN 8.088 17197 921 1.48 Cast RUBDCANTHIN 8.550 0 0 0 NF CANTHAXANTHIN 9.900 0 0 0 NF CITRAXANTH1N 11,143 7630 481 0.66 Resolved B CRYPTOXANTHIN 12.759 100344 5656 8.62 Resolved LYCOPENE 16,350 0 0 0 NF ünidcntOOló 18,783 129299 6123 11.1 1 Cast B CAftßTENE 19,418 15731 1033 1.35 Fade Figure 2 was subjected to the following conditions gave the following data Non-transesterified paprika Component TA (min)? Area Height - "Type of picid UpidenlOOOl 4,198 12917 1238 - Unident0002 4,466 31213 2498 UnidenlOOOS 6,086 36925 2042 Umdent0004 6,192 43531 3157 CAPSANTHIN 6,640 26476 1523 UnidentOOOó 6,919 13869 2208 Unident0007 7,025 47360 4396 LUTEIN K1 7,900 0 0 ZEAXANTHIN 8,000 0 0 RUBDCANTHIN 8,550 0 0 CANTHAXANTHIN 9.900 0 0, CITRAXANTHIN 11,000 0 0, B CRYPTOXANTHIN 12,792 48661 2698; Unid_nt0014 13,559 21569 873! • UnidenlOOlS 15,054 74757 2979 LYCOPENE 16,096 46150 1463 - • • Unident0017 16.647 37968 1994 UnidentOOIß 17.747 28329 1296 1 • " Unident0019 18 362 21985 1169 1 '•' B CAROTENE 18.876 183451 8834 1. ' «° Unident0021 19,526 21641 1410 1 »' Umdent0022 20,283 6424 585 t ' Unident0023 23.517 45665 2343 3.11 Fade Unident (K) 24 24.462 20179 669 1.37 Fade Unident0025 25.368 126787 5500 8.63 Fundidc Unident0026 26.136 66975 2185 4.56 Fade Unident0027 27.122 145853 6723 9.93 Fade Unident0028 27.882 70164 2310 4.78 Fade Unidem0029 28.833 100228 4103 6.82 Fade U ?? ident0030 29.594 40246 1614 2.74 Fade Urudent0031 30.509 32888 1624 2.24 Fade Urudent0032 30.732 35873 2197 2.44 Fade Unident0033 31.248 18710 891 1.27 Fade U? Üdent0034 31.833 19004 1321 1.29 Fade Unident0035 32.239 17231 1315 1.17 Fade Un_dent0036 32.814 4583 399 0.31 Fade Unident0037 33.190 17009 1279 1.16 Fade Unident0038 33.604 5286 525 0.36 Cast Totais 1468907 75361 100.00 Clearly, untreated paprika has a large number of esters in the 23-25 area that is not present in the treated material. In both treated and untreated plant materials, the 'capsantin retains the high peak in the range of 6-7 Experiment 5 Chicken test of the pigmentation effect of the calendula meal prosed in situ by the transesterifisation Warren SEX's young hens SEX -SAL-LINK were divided into three identical groups. There are groups of 7 cages that contain 3 hens each. Chickens have access to food and water all the time. All three groups were given the carrier food (pigment free feed) for 3 weeks as a control for the experiment. Group A was fed with ORO G1OTM a commercially available free form of lutein available from Industrias Kemin, Inc. Deis Moines, IA. Group B will be fed with the new invention made in accordance with the process of experiment one (7.5 mg of lutein activity per kg of feed). Group C will be fed with the new invention made according to the process of experiment one (15.0 mg of lutein activity per kg of feed). The experiment was run for 28 days. The feeding of the material requires 3 to 5 days before the effects on the solor of the yolk are visible. The color of the yolk is uniform and consistent after approximately 21 days of ingestion. Ten (10) randomly chosen eggs placed by sada group on day 0, 2, 4, 8, 16 and day 28 were broken and the color of the egg yolk was measured and the amount totaJ. of the admitted food was determined on day 28. On days 0, 16 and day 28 the yolk of the egg of 3 eggs chosen at hoisting placed by each group had to be mixed and weighed, followed by extractions and analysis of the yolk of the egg of sada group. The relationship between the color of the egg yolk and the xantofi mg / kg of feed was calculated and the ratio between the amount of xanthophylls in the egg yolk and the xanthophyll food. The result of this study was demonstrated in Figure 5, 6. Figure 5 shows the Roche abate scale for days for the color of the egg yolk of the present invention and the commercial product. Figure 6 shows the mg / kg of xanthophylls in the egg yolks during the days of the three treatments. The results show that the plant material will provide more pigment and color at the 15 mg level, then it will be the commercial product. The 15 mg level seems to be a more effective level, than the 7.5 mg level. The lower level will provide less egg yolk solder even though the egg appears, have more xanthophylls from the plant material in the yolk than the comersial product.

Claims (27)

1. CLAIMS 1. An improved plant material made from a natural plant material containing at least some non-free-living xanthophylls, characterized in that it comprises: plant materials containing in situ less than free-form xanthophylls, and more than free form xanthophylls wherein the free form of xanthophylls in situ in the improved plant material has increased beyond the amount in the material; natural plant.
2. 2. The improved plant material according to claim 1, sarasterized because the plant material is flowers.
3. 3.: _1 improved material of sonformidad plant are the reivindisasión 1, sarasterizado because the plant material is petals.
4. 4.: Improved plant material of sonification with claim 1, characterized in that the flowers are Tagetes erectéi.
5. 5. The improved plant material of conformity is claim 1, characterized in that the xanthophyll is lutein.
6. 6. The improved plant material according to claim 1, characterized in that the non-free form of xanthophyll is the fatty acid esters.
7. 7. The improved plant material according to claim 6, characterized in that the free form of Xantofila is non-acylated lutein.
8. 8. The improved sonicity plant material is reivir.disasión 1, characterized in that the improved plant material contains at least 5% more free-form xanthophylls, then performs the natural plant material in situ.
9. 9. An improved animal feed composition comprising: vitamins and minerals; a source of carbohydrates; and, improved plant material containing in situ, plus free form xanthophylls, then the natural in situ amount of xanthophylls in the plant material that the improved plant material was formed from, wherein the xanthophylls are more bioavailable to an animal fed with the animal's food.
10. 10. A feed according to claim 9, sarasterized because the improved plant material is flowers.
11. 11. A food according to claim 10, characterized in that the flowers are marigolds.
12. 12. A feed according to claim 9, characterized in that the animal feed is designed by the nutritional requirements of poultry.
13. 13. A feed according to claim 12, characterized in that the poultry are chickens.
14. 14. A food according to claim 12, characterized in that the poultry, fed with the animal feed, makes evident the bioavailability Ldad having increased pigmentation from the free form consumption of the xanthophylls.
15. 15. A food according to claim 9, characterized in that the xanthophylls are lutein.
16. 16. An improved plant material method, made from a natural plant material containing at least some xanthophylls in a non-free form, because of the steps of: treating the natural plant material in situ with a solvent; add a base sapaz of transesterifisasión of xantofilas of non-free form to the xantofilas of free form; neu.ralize the reaction wherein the formation of the improved plant material has more free-form xanthophylls, than the natural plant material.
17. 17. The method of compliance is claim 16, characterized in that it includes the step of drying the improved plant materials to remove any solvent.
18. 18. ' The method of compliance with the claim 16, characterized in that the plant material is flowers.
19. 19. The method according to claim 16, characterized in that the flowers are Tagetes erecta.
20. 20. The method according to claim 16, characterized in that the solvent is an alcohol.
21. 21. The method according to claim 20, characterized in that the alcohol has one to four carbons.
22. 22. The method according to claim 16, characterized in that the solvent is an alcohol.
23. 23. The method according to claim 16, characterized in that the alcohol is selected from the group consisting of methanol, ethanol, isopropyl alcohol.
24. 24. The method of compliance with the claim 16, characterized in that the base is selected from the group consisting of potassium hydroxide, potassium sorbate, NaOMe, animal liver lipase, NaOEt, KOMe, KOEt, Na2C03, K2C03.
25. 25. The method according to claim 16, characterized in that the step of neutralizing includes the use of a Lewis acid.
26. 26. The method according to claim 25, characterized in that the Lewis acid is Phosphoric acid.
27. 27. The method of sonification is claim 25, characterized in that the Lewis acid is selected from the group consisting of HCL, NH4CL, sulfuric acid, acetic acid, ALCL3.