WO2011082560A1 - 一种以植物油树脂为原料制备含较高玉米黄质含量的叶黄素晶体的方法 - Google Patents

一种以植物油树脂为原料制备含较高玉米黄质含量的叶黄素晶体的方法 Download PDF

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WO2011082560A1
WO2011082560A1 PCT/CN2010/071061 CN2010071061W WO2011082560A1 WO 2011082560 A1 WO2011082560 A1 WO 2011082560A1 CN 2010071061 W CN2010071061 W CN 2010071061W WO 2011082560 A1 WO2011082560 A1 WO 2011082560A1
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lutein
zeaxanthin
crystals
solvent
oleoresin
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PCT/CN2010/071061
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English (en)
French (fr)
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许新德
邵斌
晁红娟
劳学军
孙晓霞
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浙江医药股份有限公司新昌制药厂
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Priority to EP10841903.7A priority Critical patent/EP2522655B1/en
Priority to US13/521,044 priority patent/US8871984B2/en
Publication of WO2011082560A1 publication Critical patent/WO2011082560A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C403/00—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone
    • C07C403/24—Derivatives of cyclohexane or of a cyclohexene or of cyclohexadiene, having a side-chain containing an acyclic unsaturated part of at least four carbon atoms, this part being directly attached to the cyclohexane or cyclohexene or cyclohexadiene rings, e.g. vitamin A, beta-carotene, beta-ionone having side-chains substituted by six-membered non-aromatic rings, e.g. beta-carotene
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
    • C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/16—Systems containing only non-condensed rings with a six-membered ring the ring being unsaturated

Definitions

  • the invention relates to the preparation of carotenoids, in particular to a method for preparing lutein crystals containing higher zeaxanthin content by using vegetable oleoresin (especially marigold oleoresin) as raw material.
  • Carotenoids are a class of yellow to red substances that are widely found in nature. Different kinds of carotenoids exist in different fruits and vegetables, such as ⁇ -carotene in carrots, lutein in marigold flowers, zeaxanthin in strawberries, lycopene in tomatoes, and capsanthin in peppers. And the content of capsicum pigment is higher. In some dark fruits and vegetables, egg yolks, some fish shells, birds, seaweeds and bacteria, there are more carotenoids, of which the content of lutein is also higher.
  • carotenoids may be divided into two sub-categories, namely, lutein or oxygenated carotenoids with relatively strong polarity, such as lutein, zeaxanthin, astaxanthin, etc., and non-polar Sexual hydrocarbon carotenoids, such as beta-carotene, lycopene, and the like.
  • lutein or oxygenated carotenoids with relatively strong polarity, such as lutein, zeaxanthin, astaxanthin, etc.
  • non-polar Sexual hydrocarbon carotenoids such as beta-carotene, lycopene, and the like.
  • These two subclasses of carotenoids contain at least nine conjugated double bonds. These conjugated double bonds not only impart color characteristics to carotenoids, but also have strong antioxidant activity in disease control. They can prevent or prevent diseases like cancer, arteriosclerosis, cataracts, pigmentation and other diseases.
  • carotenoids can limit the oxidative destruction of free radicals due to their high ability to sca
  • Lutein and zeaxanthin are the only carotenoids found in the spotted area of the human retina (the macula), which is closely related to human visual acuity (Bone Et Al. Invest. Ophthamal. Vis. Sci. 34: 2033-2040, 1993). Frequent consumption of fruits and vegetables rich in lutein and zeaxanthin can reduce the risk of age-related degraded disease by 43% (Seddon) Et al.J. Am. Med. Assoc.
  • the structural formulas of lutein and zeaxanthin are as follows, they are isomers, the only difference between zeaxanthin and lutein is the position of the double bond on one (instead of two) end rings, the former on both end rings
  • the position of the double bond is symmetrical, while the latter is asymmetric, that is, the entire linear part of each molecule of lutein and zeaxanthin is a conjugated structure with alternating double bonds and single bonds in corn
  • the conjugated structure continues to the first bond on the two terminal rings, but lutein has a lower degree of conjugation because the double bond on one of its end rings does not form a fully conjugated structure.
  • Correct alignment is precisely the structural difference of this molecule, which causes a certain difference in functional properties between lutein and zeaxanthin.
  • Lutein (Molecular formula: C 40 H 56 O 2 molecular weight: 568.85 )
  • lutein and zeaxanthin are widely distributed in nature. They are mainly found in higher plants, algae, fish, shells and bacteria, and in organisms they often exist as esterified forms.
  • marigold is a good source of lutein and zeaxanthin
  • 100 g marigold flowers often contain about 2g of lutein, the main one is lutein, accounting for 90% Above, the rest are zeaxanthin and a small amount of other carotenoids.
  • lutein accounts for a larger proportion than zeaxanthin, but in corn, the amount of zeaxanthin is greater than that of lutein.
  • a method for obtaining a large amount of lutein crystals which is economical and easy to mass-produce is obtained by extracting, separating and purifying from natural resources.
  • Plant xanthophylls are often in the form of a monoester or diester with a 12 -C 18 long-chain fatty acids such as esterified some C myristic acid, oleic acid, linoleic acid and palmitic acid, or the like is present.
  • Lutein esters are generally extracted from plants using organic solvents, preferably from marigold flowers, marigolds and other dark green vegetables, and these organic solvents are relatively easy to separate themselves.
  • the marigold flower bead extract (marigold oleoresin) is a good source of lutein ester, and the other carotenoids are relatively low in content. After hydrolysis under alkaline conditions, the lutein crystals are released, and the lutein crystals can be further purified after washing off the fatty acid salt obtained by soaping.
  • zeaxanthin due to the presence of a conjugated double bond in the molecular structure, zeaxanthin has stronger antioxidant activity than lutein, which may play a more important role in human eye health.
  • Upper, 80 Some studies in the middle and late chronological studies have also proved that the zeaxanthin is mainly in the cell at the center of the human eye macular, and concentrically leaves the concave portion and approaches the outer periphery of the macula, the amount of zeaxanthin gradually decreases, and lutein The amount is gradually increased, and lutein is the main yellow pigment on the periphery of the macula.
  • the ratio of lutein to zeaxanthin in the raw material of lutein is 10-12:1. In the human blood, the ratio is about 3-5:1. In the periphery of the retinal macula, the ratio of lutein to zeaxanthin is 3:1. In the center of the macula, this value is completely reversed, and the ratio of lutein to zeaxanthin becomes 1:3.
  • the lutein can be partially heterologous to form zeaxanthin during the preparation of lutein, it can be ensured that the obtained product contains both lutein and zeaxanthin, and the required ratio is maintained, so that the product is When applied, it is possible to add only one carotenoid dosage form containing a certain proportion of lutein and zeaxanthin, which will bring a lot of convenience to the subsequent application process.
  • the marigold oleoresin after saponification is washed with water at a lower temperature in U.S. Pat. No. 5,382,714, and crystallized at a low temperature in a mixed solvent to separate and purify lutein.
  • This purification process is not only time consuming, but also uses a chlorine-containing organic solvent, and the obtained product is not suitable for use in foods and medicines.
  • the proportion of lutein and zeaxanthin in the product did not change significantly compared to the raw material.
  • U.S.Pat.No. 5,648,564 A method for separating lutein crystals is disclosed by first dissolving lutein diester in a propylene glycol solution of marigold oleoresin and recrystallizing. This process also has several disadvantages: First, due to the high viscosity of propylene glycol, the temperature is required to be high during saponification and subsequent processing.
  • U.S.Pat.No. 6,262,284 It describes the extraction and solubilization of carotenoids from dried flowers of marigold with tetrahydrofuran. In the process, a large amount of organic solvents are used, which are unfavorable to the stability of lutein, which may lead to product peroxidation. And caused by degradation.
  • U.S.Pat.No.6.329,557 A method for extracting lutein crystals from an industrial scale of marigold oleoresin is disclosed.
  • the disadvantage of this process is the use of a large amount of organic solvents such as n-hexane, ketones, etc., which are not suitable for use in foods.
  • USPat.No. 7,485,738 uses lutein as a raw material, and isomerized to obtain high-purity meso-zeaxanthin under strong organic base catalysis.
  • the zeaxanthin crystal obtained in this process contains no or very A small amount of lutein.
  • the methods described in the above patents have the following disadvantages: 1) Some toxic organic solvents are used in the process. These solvents are difficult or impossible to remove completely, resulting in the product lutein or zeaxanthin crystals not suitable for human consumption; 2) Or because the viscosity of the organic solvent used is large, the operation in the separation and purification process is difficult, in order to obtain a high content of crystal, a multi-step crystallization process is required, which is not suitable for industrial production; 3) Or the yield of carotenoid crystals is low, because the process involves multiple steps, resulting in low product yield, only 50% Left and right, especially using marigold flower as raw material, through the soapy reaction and separation and purification to obtain lutein crystals, and then transforming lutein crystals by isomerization reaction to obtain a mixture of zeaxanthin and lutein, the yield is Lower; 4) Or the concentration of a single lutein or zeaxanthin in the product is relatively high, and the reaction process cannot be controlled as
  • the present invention provides a method for preparing a lutein crystal containing a high zeaxanthin content by using a vegetable oleoresin as a raw material, and which can conveniently and effectively adjust the lutein crystals.
  • the purpose of zeaxanthin content is a method for preparing a lutein crystal containing a high zeaxanthin content by using a vegetable oleoresin as a raw material, and which can conveniently and effectively adjust the lutein crystals.
  • the purpose of zeaxanthin content is zeaxanthin content.
  • the vegetable oleoresin is mixed with a food-grade alcohol solvent to form a uniform free-flowing mixture by stirring, and then the mixture is lyophilized in an alkaline environment at 40-85 ° C for 3-5 hours;
  • the resulting reaction solution reduces the volume concentration of the solute in the solution to 10-50%, the amount of deionized water and alcohol solvent is 2-10 times and 0.5-2 times of the weight of the raw vegetable oleoresin, then the temperature is raised to 60-75 ° C, slow stirring 0.5-2.0 In an hour, the carotenoid forms a granular crystal and precipitates;
  • the filter cake is rinsed with anhydrous ethanol once before drying, and then dried under vacuum or freeze-dried to obtain lutein crystals, so that the final loss on drying is less than 5%.
  • the vegetable oleoresin used in the present invention is an extract of marigold flower, marigold flower, spinach, strawberry, broccoli, corn, and cabbage, and among these raw materials, the content of lutein in the marigold flower extract is relatively high, which is a priority.
  • this oleoresin contains 5-30% lutein diester and a small amount of other carotenoids, such as all-trans zeaxanthin, ⁇ - and ⁇ -cryptus yellow. Quality and beta-carotene.
  • UV-visible spectrophotometric analysis and HPLC analysis showed that the final product contains 75-95% carotenoids, including 15%-95% all-trans lutein, 85%-5% zeaxanthin, and possibly There are also 0.1-1.0% of its geometric isomers, and less than 1.0% of other carotenoids. These traces of other carotenoids are completely unnecessary to be harmful, as they are themselves a dietary source and are found to be much higher in human serum than lutein.
  • the final crystalline product produced by this process does not contain toxic organic solvent residues and other toxic compounds, and is suitable for human consumption.
  • step a) of the present invention Generally, one part by weight of vegetable oil resin is dissolved in 0.5-2.0 volumes of food-grade alcohol solvent, and by stirring, lutein ester and other impurities such as wax, resin, other carotenoids and pigments are dissolved or dispersed. A uniform solution is formed in the solvent; an alkali solution (such as NaOH, KOH, sodium methoxide or sodium ethoxide) is slowly added dropwise to the uniformly dispersed solution for saponification, and the amount of alkali required is lutein diester in the oleoresin.
  • an alkali solution such as NaOH, KOH, sodium methoxide or sodium ethoxide
  • lutein, zeaxanthin and other carotenoids are freed, while fatty acids in vegetable oleoresin (such as myristic acid, palmitic acid, stearic acid, etc.) and sodium Or potassium ions form sodium soap or potassium soap, respectively, and the food-grade alcohol solvent used is methanol, ethanol, isopropanol, propanol or the like, preferably ethanol.
  • an organic solvent is added to increase the solubility of the free carotenoid therein, and the organic solvent is used in an amount of 1.0 to 3.0 times by weight of the vegetable oleoresin , and the organic solvent may be used.
  • It may be an alcohol solvent such as ethanol, isopropanol, propanol, propylene glycol or the like, and may also be an ester solvent such as ethyl acetate, ethyl isobutylate or the like, or other organic solvents such as dimethyl sulfoxide or dichloride.
  • the emulsifier used may be selected from Tween series emulsifiers such as Tween-60, or Span series.
  • Emulsifiers, such as Span-40, etc. the amount of emulsifier is 0.4-1.0 times the mass of lutein crystals in vegetable oil resin, so that a certain amount of strong base is added dropwise (the amount of strong base is lutein diester in vegetable oleoresin) After 0.1-1.0 times the mass, no additional steps are required to precipitate the lutein crystals to directly form part of the lutein epimer to form zeaxanthin.
  • the ratio of lutein to zeaxanthin in the reaction product can be monitored by high performance liquid chromatography after sampling. According to the ratio of lutein to zeaxanthin, the reaction time and temperature can be adjusted. The reaction time is selected from 0.5 to 6.0 hr, and the temperature is selected from 60 to 90 °C.
  • step e) of the present invention by a conventional separation process such as centrifugation, filtration, pressure filtration, etc.
  • the resulting crystals are separated, and the solution may be diluted with an appropriate amount of hot water in order to further reduce the concentration of the crystals before the separation and crystallization to facilitate the subsequent separation operation.
  • impurities such as fatty acids, salts, soaps, water-soluble chlorophyll and flavonoids are transferred to the mother liquor, and crystals such as lutein and zeaxanthin are left in the filter cake.
  • the purity of the final product, the ratio of lutein and zeaxanthin, and the yield of carotenoid crystals depend not only on the amount of solvent added during the soaping process, but also the amount of the alcohol solvent added during the dilution, and the isomerism.
  • the solubility in the solution will be much lower, but to convert it to zeaxanthin, it must be dissolved to participate in the reaction. Therefore, after the completion of the soaping reaction, a certain amount of organic solvent is added before the start of the isomerization reaction.
  • the concentration of the oleoresin before the soapy reaction makes the carotenoid oleoresin complete in a short period of time, improves the carotenoid content in the final product, and on the other hand ensures that the reactants are free during the isomerization reaction.
  • the fully dissolved carotenoid crystals make the isomerization reaction controllable.
  • the free carotenoid crystals are obtained by dissolving the vegetable oleoresin, but before the crystals are dissolved in the organic solvent to cause the isomerization reaction, the concentration of the alkali is not too high, otherwise some of the carotenoid oleoresin and the formed carotenoid crystals will It is lost by high concentration of alkali oxidation, and even carbonization occurs in severe cases. This phenomenon can be avoided by adding different amounts of alkali in stages. The same is true for the purpose of adding a strong base in the late isomerization of lutein to produce zeaxanthin.
  • the process of refining the aqueous alcohol solution before separating the final carotenoid crystal also has an effect on the content and yield of the product. Excessive addition of alcohol during the dilution process will reduce the yield of the product, and too little addition will increase the difficulty of operation. The time for precipitation crystallization should not be too short, otherwise the yield will be lowered.
  • the advantages of the invention are as follows: 1) using a vegetable oil resin as a raw material, through a two-step reaction, that is, a soaping reaction and an isomerization reaction, a crystal containing lutein and zeaxanthin is obtained in one step, and the ratio of lutein to zeaxanthin in the crystal is determined according to Need to be adjusted by controlling the reaction conditions, which facilitates subsequent product application, effectively avoids the inconvenience of purchasing, storage and operation caused by mixing lutein and zeaxanthin respectively; 2) passing a series of conditions Optimized to obtain a mixture of lutein and zeaxanthin crystals in a high yield, without additional manipulation to purify lutein crystals during the reaction, the isomerization reaction is directly carried out, which is higher than the current process.
  • Marigold oleoresin (total lutein content 15.2%) is mixed with 2000mL of isopropanol, heated to 40 ° C, stirred until a uniform flow of the solution is formed, and 405mL of 45% NaOH solution is slowly added dropwise while stirring, the dropping time is After 60 minutes, the solution was hydrolyzed at this temperature for 5 hours.
  • reaction solution was heated to 60 ° C, 2400 mL of an isopropanol solution was added, and 80 g of an emulsifier Tween-80 was stirred for 0.5 hr to make it uniformly mixed.
  • 240 mL of 37% CH 3 ONa solution was slowly added dropwise, and the dropping time was 1.0 hr, while the temperature was raised to 70 ° C, and the reaction was stirred for 0.5 hr.
  • the product does not contain toxic organic solvents and is suitable for use in the form of nutritional supplements and food additives.
  • the application form of the crystal may be an oil suspension (mixed with vegetable oil), beads (microcapsules obtained by spray condensation), dry powder (microcapsules obtained by spray drying), and the like.
  • Marigold oleoresin 500g Marigold oleoresin (total lutein content 14.5%) is mixed with 250mL of propanol, heated to 85 ° C, stirred until a uniform flow of the solution is formed, and 210mL of 50% KOH solution is slowly added dropwise with stirring, the dropping time is 60 In minutes, the solution was hydrolyzed at this temperature for 3 hours.
  • reaction solution was cooled to 60 ° C, and 1500 mL of ethyl acetate solution was added to emulsifier Span-40. After 65 g, the mixture was stirred for 0.5 hr to make it uniformly mixed. 100 mL of a 50% sodium ethoxide solution was slowly added dropwise, and the dropping time was 1.0 hr, while the temperature was raised to 80 ° C, and the reaction was stirred for 6.0 hr.
  • the product does not contain toxic organic solvents and is suitable for use in the form of nutritional supplements and food additives.
  • the application form of the crystal may be an oil suspension (mixed with vegetable oil), beads (microcapsules obtained by spray condensation), dry powder (microcapsules obtained by spray drying), and the like.
  • Marigold oleoresin (total lutein content 14.5%) is mixed with 2500mL of ethanol, heated to 75 ° C, stirred until a uniform flow of the solution is formed, and 380mL of 50% KOH solution is slowly added dropwise with stirring, the dropping time is 60 minutes. The solution was hydrolyzed at this temperature for 4 hours.
  • reaction solution was cooled to 60 ° C, and 1200 mL of propanol solution was added to emulsifier Tween-60. After 60 g, the mixture was stirred for 0.5 hr to make it uniformly mixed. 100 mL of a 37% sodium methoxide solution was slowly added dropwise, and the dropping time was 1.0 hr, while the temperature was raised to 80 ° C, and the reaction was stirred for 3.0 hr.
  • Example 4-7 Project / step Example 4
  • Example 5 Example 6
  • Example 7 The amount of raw materials (g) and the content of lutein 300g/14.5% 1500g/13.6% 480g/15.4% 600g/14.6% Solvent type and quantity in soapy reaction Methanol / 600ml Ethanol / 1200ml N-propanol / 500ml Ethanol / 800ml Type and amount of base catalyst in soapy reaction 37% sodium methoxide solution / 118ml 50% sodium ethoxide solution / 816ml 50% KOH ethanol solution / 1780ml 45% NaOH solution / 400ml Type and quantity of solvent added before isomerization Ethyl acetate / 300ml Ethanol / 1500ml Isopropyl alcohol / 1200ml Isobutyl acetate / 1800ml Type and quantity of emulsifier added before isomerization Tween-60/ 17.5g Span-40/ 204g T

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Description

一种以植物油树脂为原料制备含较高玉米黄质含量的叶黄素晶体的方法 技术领域
本发明涉及类胡萝卜素的制备，具体地说是一种以植物油树脂（特别是万寿菊油树脂）为原料制备含较高玉米黄质含量的叶黄素晶体的方法。
背景技术
类胡萝卜素是自然界中广泛存在的一类黄色到红色的物质。在不同的水果和蔬菜中，存在着不同种类的类胡萝卜素，比如胡萝卜中β-胡萝卜素，万寿菊花中叶黄素，草莓中玉米黄质，番茄中番茄红素，辣椒类植物中辣椒红色素和辣椒色素的含量都较高。而在一些深色水果和蔬菜、蛋黄、一些鱼类甲壳、鸟类、海藻和细菌中存在较多的类胡萝卜素，其中叶黄素类的含量也较高。
近年来的一些动物和人体试验表明了类胡萝卜素的有益功效。总体上类胡萝卜素可能分为两个亚类，即极性相对较强一点的叶黄素类或含氧类胡萝卜素类，如叶黄素、玉米黄质、虾青素等，以及非极性的碳氢类胡萝卜素类，比如β-胡萝卜素、番茄红素等。这两个亚类的类胡萝卜素中都包含着至少九个共轭双键，这些共轭双键不仅赋予了类胡萝卜素的颜色特征，也使其在疾病防治中具有较强的抗氧化功能，它们能阻止或预防像癌症、动脉硬化、白内障、色斑退化及其它类疾病。而且由于类胡萝卜素的清除高效活性氧自由基及防止自由基产生的能力，它们能限制自由基的氧化破坏作用。
在所有的类胡萝卜素中，由于其潜在的预防一种与年龄相关的色斑退化疾病（ARMD）的能力，叶黄素和玉米黄质越来越引起科学家和公众的关注。叶黄素和玉米黄质是唯一存在于人类视网膜中斑点区域(视黄斑)的类胡萝卜素，而此区域与人的视觉敏锐性密切相关（Bone et al.Invest.Ophthamal.Vis.Sci.34:2033-2040,1993）。经常食用富含叶黄素和玉米黄质的水果和蔬菜能使患老年色斑退化疾病的风险减少43%（Seddon et al.J.Am.Med.Assoc.272:1413-1420,1994），而且，目前这些化合物在预防老年色斑退化疾病上的代谢途径已经明了。美国食品和药品监督管理局也认为叶黄素和玉米黄质“一般认为安全”（GARS）。因此，这些类胡萝卜素类能单独或与其它物质一起作为营养补充剂和食品着色剂使用，也可应用于临床预防老年色斑退化和癌症等。
叶黄素和玉米黄质的结构式分别如下，它们是同分异构体，玉米黄质和叶黄素唯一的差别在于一个（而不是两个）端环上双键的位置，前者两个端环上双键的位置是对称的，而后者是不对称的，也就是说，叶黄素和玉米黄质每一个分子的整个直链部份都是共轭结构，具有交替的双键和单键，在玉米黄质分子中，共轭结构延续至两个端环上的第一个键，但叶黄素的共轭程度更低，因为它的端环之一上的双键未形成完全共轭结构的正确排列，正是这一分子结构上的差异，造成了叶黄素和玉米黄质在功能性质上有一定的差别。
Figure PCTCN2010071061-appb-C000001
叶黄素（分子式： C40H56O2 分子量： 568.85 ）
Figure PCTCN2010071061-appb-C000002
玉米黄质（分子式： C40H56O2 分子量： 568.85 ）
作为一种天然色素，叶黄素和玉米黄质在自然界中分布很广，它们主要存在于高等植物、藻类、鱼类、壳类和细菌中，并且在生物体内它们往往以酯化物形式存在，在这些物质中，万寿菊花是一种良好的叶黄素和玉米黄质的来源，100 g 万寿菊鲜花中往往存在 2g 左右的叶黄素类物质，其中主要的是叶黄素，占 90% 以上，其余为玉米黄质和少量的其它类胡萝卜素。和万寿菊花中一样，在其它种高等植物及藻类来源中，相对于玉米黄质，叶黄素占的比例较大，只是在玉米中，玉米黄质的量要比叶黄素的量大。另外，从分子结构中也可看出，叶黄素和玉米黄质中存在立体异构现象，来源不同，其立体异构构成也不一样，比如在植物原料中 (3R,3'R,6'R)- 叶黄素或 (3R,3'R)- 玉米黄质是主要的，但在动物来源中，如在鱼类和壳类，叶黄素是以 (3R,3'R,6'R)- ， (3R,3'R,6'S)- ，和 (3R,3'S,6'S)- 的形式。
由于化学合成叶黄素类涉及到多步反应，费时费力，成本很高。比较经济且易于大规模生产而得到大量叶黄素晶体的方法是从天然资源中提取、分离和纯化得到。
许多蔬菜和水果，比如菠菜、椰菜、甘蓝和玉米等，含有较多的叶黄素，但万寿菊花和金盏花是叶黄素最丰富来源，当然，这些植物中还存在其它类类胡萝卜素。植物中的叶黄素类往往是与一些C12-C18 长链脂肪酸如肉豆蔻酸、油酸、亚油酸和棕榈酸等酯化形成单酯或双酯的形式存在。
一般使用有机溶剂从植物，优先从万寿菊花、金盏花和其它深绿色蔬菜中提取叶黄素酯，而且这些有机溶剂本身又比较容易分离出去。万寿菊花珠粒提取物（万寿菊油树脂）是一种很好的叶黄素酯来源，并且其中其它类类胡萝卜素的含量相对不高。在碱性条件下水解后，叶黄素晶体就游离出来，将皂解得到的脂肪酸盐洗掉后，叶黄素晶体可进一步得到纯化。
同时，如前所述，由于分子结构式中多存在一个共轭双键，使得玉米黄质比叶黄素有更强的抗氧化活性，其在人眼健康中可能取到了更重要的作用， 事实上， 80 年代中后期的一些研究也证明了在人眼视黄斑正中心的小区内主要是玉米黄质，同心地离开凹部而向黄斑的外周边靠近，则玉米黄质的量逐渐减少，叶黄素的量随之渐增，在黄斑***，叶黄素是主要的黄色色素。
这一点也可从自然界和人体组织不同部位中叶黄素和玉米黄质比例的变化中看出，叶黄素的原料万寿菊花中叶黄素和玉米黄质的比例为 10-12:1 左右，在人体血液中这一比例大概为 3-5:1 ，在视网膜视黄斑***，叶黄素与玉米黄质的比例为 3:1 ，而在视黄斑中心，这一数值完全颠倒过来，叶黄素与玉米黄质的比例变成了1:3 。而且最近的研究发现，在视黄斑中心区域，玉米黄质中的一种异构体内消旋玉米黄质 (3R,3'S, meso)-zeaxanthin 占有较大的比例，越往***，这种比例会越小。越来越多的证据表明视黄斑中的内消旋玉米黄质是叶黄素通过差向异构化转位得到的，因为在自然界、人体血液及人体其它组织中几乎检测不到玉米黄质的这种异构体。
叶黄素和玉米黄质在人眼中不同部位分布比例的变化恰恰说明了玉米黄质特别是内消旋玉米黄质在人体眼睛健康中重要而又独特的作用。实际上许多人体试验也证明了叶黄素和玉米黄质复合使用时会起到更好的作用，所以目前市场上的大多数含叶黄素的护眼产品中往往额外添加了一定量的玉米黄质，也就是在应用配方中按一定的配比分别加入叶黄素和玉米黄质。但叶黄素和玉米黄质的分别加入必将会对原料的采购、生产过程操作、产品质量控制带来许多不必要的麻烦。如果能在叶黄素制备过程中同时能让部分叶黄素差向异体化形成玉米黄质，就能保证得到的产品中同时包含有叶黄素和玉米黄质，并保持需要的比例，这样在产品应用时就可以只加入一种同时含某种比例的叶黄素和玉米黄质的类胡萝卜素剂型，这将会给后续的产品应用过程带来很多方便。
目前也有一些专利和文献涉及到从万寿菊珠粒中大规模分离叶黄素晶体或者通过叶黄素差向异构化转位得到玉米黄质晶体的方法。这些文献的目的一般是得到较纯的叶黄素或玉米黄质晶体形式，并涉及到多个分离步骤。
U.S.Pat.No.5,382,714中在较低的温度下通过水洗皂解后的万寿菊油树脂，并在混合溶剂中低温下结晶，从而分离和提纯叶黄素。此纯化过程不仅费时，而且使用到含氯有机溶剂，得到的产品不适于食品和药物上使用。产品中叶黄素和玉米黄质比例与原料相比并没有发生大的变化。
U.S.Pat.No.5,648,564 公开了一种分离叶黄素结晶的方法，先在万寿菊油树脂的丙二醇溶液中皂解叶黄素二酯，再重结晶。这个过程也有几个缺点：首先，由于丙二醇的粘度较大，在皂化及随后的处理过程中要求温度较高，整个体系在 70 ℃ 以上的温度要保持10小时左右，这对叶黄素的稳定性显然不利，叶黄素的顺反异构体也会发生变化，而且，随后的分离过程如离心或过滤也比较困难；其次，叶黄素的收率较低，只有59%左右，产品中叶黄素的含量也不高。
U.S.Pat.No.6,262,284 中描述了用四氢呋喃同时从万寿菊干花中提取和皂解类胡萝卜素，在此过程中要用到大量的有机溶剂，这些溶剂对叶黄素的稳定性不利，有可能会导致产品由于过氧化而引起的降解。
U.S.Pat.No.6.329,557 揭示了一种从万寿菊油树脂中工业化规模提取叶黄素结晶的方法。这个过程的缺点是使用到大量的有机溶剂正己烷、酮类等，这些溶剂不适于在食品中使用。
U.S.Pat.No.6,380,442 报道了一种从植物中分离类胡萝卜素的方法，该方法在工业生产上也不是很有吸引力，因为生产过程中要用到大量的水（至少为原料量 30 倍以上），而且操作比较困难。
U.S.Pat.No.6,743,953 使用有机溶剂从万寿菊油树脂中分离和纯化叶黄素类。此过程中用到异丙醇、乙酸乙酯、正己烷、丙酮和甲醇等多种有机溶剂，操作繁杂，有机溶剂消耗大，收率低。所以此方法也不适于工业化生产。
U.S.Pat.No.7,271,298 公开了一种用无水乙醇为溶剂，通过简单的工艺高收率得到高含量叶黄素晶体的方法，此方法得到的晶体也没有考虑到提高其中玉米黄质的比例。
U.S.Pat.No.5,780,693(CN1,178,787A) 设计了一条由叶黄素为原料生产玉米黄质的路线，主要是以二甲基亚砜或与饱和烷烃和／或芳烃有机溶剂的混合物作溶剂，以碱金属氢氧化物作催化剂转位叶黄素生产玉米黄质，此工艺中产品的收率相当低 ( 重复试验显示，依此方法得到的产品收率不足 30%) 。而且，反应过程使用到了正己烷、正庚烷、二氯甲烷、甲醇等有机溶剂，使用这些有毒的溶剂生产食用级或药用级玉米黄质显然是不合适的。
U.S.Pat.No.7,485,738使用叶黄素为原料，在强有机碱催化下，差向异构化得到高纯度的内消旋玉米黄质，此工艺中得到的玉米黄质晶体不含或含非常少量的叶黄素。
总之，以上这些专利描述的方法存在以下几个缺点： 1) 过程中使用到一些有毒的有机溶剂，这些溶剂比较困难或者不可能完全去除，导致产品叶黄素或玉米黄质结晶不适于人类食用； 2) 或者由于使用到的有机溶剂粘度较大，分离和纯化过程中的操作比较困难，为了得到高含量的晶体，需要多步的结晶过程，不适于工业化生产； 3) 或者类胡萝卜素晶体的收率较低，由于过程中涉及到多个步骤，导致产品收率低，只有 50% 左右，特别是利用万寿菊花为原料，通过皂解反应并分离精制得到叶黄素晶体后，再通过异构化反应部分转化叶黄素晶体得到玉米黄质和叶黄素混合物时，收率就更低； 4) 或者产品中单一的叶黄素或者玉米黄质的浓度较高，而不能根据需要对反应过程进行控制从而达到调节产品中叶黄素和玉米黄质浓度的目的。
因此，有必要找到一种适于工业化规模生产高纯度叶黄素和玉米黄质的方法，在此过程中用到尽量少的毒性有机溶剂和操作步骤，但有较高的收率，并且能根据需要对反应参数进行控制，从而达到调节产品中叶黄素和玉米黄质浓度的目的。
发明内容
为克服上述现有方法存在的缺陷，本发明提供一种以植物油树脂为原料制备含较高玉米黄质含量的叶黄素晶体的方法，并且其能达到便利且有效地调节叶黄素晶体中玉米黄质含量的目的。
为此，本发明采用的步骤如下：
a ) 采用 含叶黄素二酯 的植物油树脂与食品级醇类溶剂混合，通过搅拌，形成均一的能自由流动的混合液，再在碱性环境下、40-85℃下皂解该混合液3-5小时；
b) 向 a) 所得混合液补加有机溶剂和加入乳化剂，搅拌均匀；
c) 向 b) 所得混合液中滴加强碱溶液，使得到的部分叶黄素晶体差向异构化转化成玉米黄质，根据要求的叶黄素与玉米黄质比例，对反应时间和温度进行调节 ；
d) 用去离子水和醇类溶剂组成的混合溶剂稀释 c) 所得的反应液，使溶液中溶质的体 积浓度降低至 10-50% ，去离子水与醇类溶剂的用量分别为原料植物油树脂重量的 2-10 倍与 0.5-2 倍，然后升温至 60-75℃ ，慢速搅拌 0.5-2.0 小时，使类胡萝卜素 形成颗粒状结晶 析出；
e) 采用 离心或过滤方法得到 d) 中 生成的结晶 ；
f) 用 60-85℃的热水漂洗e) 中得到的结晶2-3次，直至洗出的母液接近无色；
g) 为了使最终产品易于干燥，干燥之前结晶用无水乙醇淋洗滤饼一次，然后采用真空干燥或冷冻干燥得到叶黄素晶体，使其最终的干燥失重小于5%。
本发明所用的植物油树脂为万寿菊花、金盏花、菠菜、草莓、椰菜、玉米、甘蓝的提取物，在这些原料中，万寿菊花提取物中叶黄素类含量相对较高，是一种优先考虑的原料。基于品种、种植条件、收割时期及提取方法的不同，此油树脂含5-30%的叶黄素二酯和少量其它类胡萝卜素，如全反式玉米黄质、α-和β-隐黄质以及β-胡萝卜素等。
通过紫外-可见光分光光度分析和HPLC分析得知最终产品含75-95%类胡萝卜素类，其中包括15%-95%全反式叶黄素，85%-5%的玉米黄质，另外可能还存在0.1-1.0%其几何异构体，及少于1.0%的其它类胡萝卜素。这些微量的其它类类胡萝卜素完全不用担心有害，因为其本身也是膳食来源，并且发现人体血清中相对于叶黄素，它们的浓度要高得多。
通过这个工艺生产的最终结晶产品中不存在毒性有机溶剂残留和其它有毒化合物，可适于人类食用。
在本发明 a ) 步骤 中， 一般用一份重量的植物油树脂溶解于0.5-2.0倍体积的食品级醇类溶剂中，通过搅拌，叶黄素酯及其它类杂质如蜡、树脂、其它类胡萝卜素和色素等溶解或分散于此溶剂中形成一种均一的溶液；向此均匀分散的溶液中缓慢滴加碱溶液（如NaOH、KOH、甲醇钠或乙醇钠）进行皂化，所需的碱量为油树脂中叶黄素二酯质量的0.5-6.0倍，在此过程中，叶黄素、玉米黄质及其它类胡萝卜素类游离出来，同时植物油树脂中的脂肪酸（如肉豆蔻酸、棕榈酸、硬脂酸等）与钠或钾离子分别形成钠皂或钾皂，用到的食品级醇类溶剂为甲醇、乙醇、异丙醇、丙醇等，优选乙醇。
在本发明 b ) 和 c) 步骤 中， 补加一定量的有机溶剂以加大游离出的类胡萝卜素在其中的溶解度，有机溶剂的用量为植物油树脂重量的1.0-3.0倍体积，有机溶剂可以为醇类溶剂如乙醇、异丙醇、丙醇、丙二醇等，也可为酯类溶剂如乙酸乙酯、乙酯异丁酯等，也可以为其它有机溶剂如二甲基亚砜、二氯甲烷等；加入一定量的乳化剂使类胡萝卜素能更好地溶解，用到的乳化剂可以选自吐温(Tween)系列乳化剂如Tween-60等，也可为司班(Span)系列乳化剂，如Span-40等，乳化剂的用量为植物油树脂中叶黄素晶体质量的0.4-1.0倍，这样在补充滴加一定量的强碱（强碱的用量为植物油树脂中叶黄素二酯质量的0.1-1.0倍）后，不需要额外的步骤将叶黄素晶体析出就可以直接使部分叶黄素差向异构体生成玉米黄质。反应产物中叶黄素和玉米黄质的比例，可以通过取样后用高效液相色谱法进行监控。根据要求的叶黄素与玉米黄质比例，可以对反应时间和温度进行调节 ， 反应时间的选用范围为0.5-6.0hr，温度的选用范围为60-90℃。
在本发明 e) 步骤 中， 通过传统的分离过程如离心、过滤、压滤等将 d) 中 生成的结晶分离出来，在分离结晶之前，为了进一步减少结晶的浓度以使随后的分离操作过程易于进行，可用适量的热水稀释溶液。经过分离操作后，脂肪酸、盐、皂、水溶性叶绿素和黄酮等杂质转移到母液中，叶黄素和玉米黄质等结晶留在滤饼中。
需要说明的是，最终产品的纯度、叶黄素和玉米黄质的比例、类胡萝卜素晶体的收率不仅取决于皂解过程中溶剂的加入量和稀释过程中醇类溶剂的加入量，以及异构体反应过程中碱的滴加量、反应时间和反应温度，而且还取决于水洗前结晶的时间。
皂解反应时溶剂比例过少，会使溶液的粘度较大，不利于反应及随后的分离操作；溶剂的比例太大，会使溶液中叶黄素酯类的浓度变稀，从而不利于皂解完全，减少产品的收率，降低结晶含量。这也就是一部分有机溶剂需要在异构化反应前补加的原因，因为油树脂在所用溶剂中的溶解度相对较大，而皂解反应完成后得到的是类胡萝卜素晶体，这些晶体在有机溶剂中的溶解度会降低很多，但要使其转化为玉米黄质就必须先让其溶解参与反应，所以在皂解反应完成后异构化反应开始前补充一定量的有机溶剂，一方面不会稀释皂解反应前油树脂的浓度，使类胡萝卜素油树脂在较短时间内皂解完成，提高终产品中类胡萝卜素含量，另一方面又能保证异构化反应时反应物也就是游离的类胡萝卜素晶体的充分溶解，使异构化反应可控。
在不同的反应阶段分批加入碱也是非常重要的。在皂解植物油树脂得到游离类胡萝卜素晶体但这些晶体还没来得及溶解于有机溶剂以发生异构化反应之前，碱的浓度不能太高，否则部分类胡萝卜素油树脂和生成的类胡萝卜素晶体会被高浓度的碱氧化损失，严重时甚至发生炭化。而通过分阶段添加不同量的碱就能避免这一现象的发生。在后期异构化叶黄素生成玉米黄质过程中强碱需要以滴加的方式加入的目的也是如此。
在异构化反应前加入一定量的乳化剂对异构化反应的发生也很有好处。因为在皂解反应和异构化反应过程中需要加入一定量的碱溶液作为反应的催化剂，不可避免地带入少量的水，这些水的存在必定会极大地降低类胡萝卜素晶体在有机溶剂中的溶解度，从而减少了异构化反应的机率，而通过加入一定量的乳化剂就能将这些少量水对类胡萝卜素晶体在有机溶剂中溶解度的影响减少到最小，能加大类胡萝卜素的溶解度，从而有利于反应的发生。而且，少量乳化剂的存在也能增加碱催化剂与反应底物类胡萝卜素晶体接触的机会，使异构化反应更易进行。
另外分离最终类胡萝卜素晶体前的醇水溶液精制过程对产品的含量和收率也有影响。稀释过程中醇类加入过多，会降低产品的收率，加入过少，也会增大操作难度。沉淀结晶的时间也不能过短，否则会使收率降低。
 有益效果
本发明的优点如下：1)以植物油树脂为原料，通过连续两步反应即皂解反应和异构化反应，一次性得到含叶黄素和玉米黄质的晶体，晶体中叶黄素和玉米黄质的比例根据需要可通过控制反应条件而进行调节，方便了后续的产品应用，有效地避免产品应用时需要将叶黄素和玉米黄质分别混合带来的采购、贮存及操作上的不便；2)通过一系列条件的优化，能以较高的收率得到叶黄素和玉米黄质晶体混合物，反应过程中不需要额外的操作来纯化叶黄素晶体就直接进行异构化反应，这比目前的工艺中先得到高纯度的叶黄素晶体，再将此叶黄素晶体部分异构化以得到叶黄素和玉米黄质晶体混合物时的收率高很多；3)通过分批加入有机溶剂和碱催化剂，既能保护反应的正常发生，又能使反应时间尽量缩短且能保证反应完全，而且能减少强碱性对反应产物和底物的破坏作用；4)在差向异构化反应发生阶段通过加入一定量的乳化剂，增大类胡萝卜素晶体在有机溶剂中的溶解度，而且能有效地增加碱催化剂与反应底物的接触机会，使反应更加完全；5)由于过程中使用到的溶剂为水和低级醇类或酯类，它们粘度较低，则下步的分离操作过程相对比较容易，也没有用到额外的有机溶剂重结晶精制过程。总之，本发明所述的方法很经济并适于大规模商业化生产。
本发明的实施方式
下面结合具体实施方式对本发明作详细说明。
实施例 1
1000g 万寿菊油树脂(总叶黄素含量15.2%)与2000mL异丙醇混合，升温至40℃,搅拌直至形成一种均一流动的溶液，边搅拌边缓慢滴加405mL45%NaOH溶液，滴加时间为60分钟，在此温度下皂解反应5小时。
反应液升温到60℃,加入异丙醇溶液2400mL，乳化剂Tween-80 80g后搅拌0.5hr使其混合均匀。缓慢滴加37%的CH3ONa溶液240mL，滴加时间为1.0hr，同时升温到70℃,搅拌反应0.5hr。
反应完成后加入2000mL去离子水和200mL乙醇，稀释过程中溶液温度保持在60℃，慢速搅拌30分钟。用离心方法分离所得到的结晶，此分离过程比较容易，10分钟内操作完成。收集到的结晶用60℃热水洗2-3次直至洗出液几乎无色，最后用无水乙醇淋洗，再在40℃下进行真空干燥直至其干燥失重小于5%。
最后得到144.6g成品，其中含87.2%总类胡萝卜素（用紫外-可见光光度仪分析）,这些类胡萝卜素中含95.1%全反式叶黄素，4.8%全反式玉米黄质(用HPLC分析)，其余的为微量其它类类胡萝卜素。总类胡萝卜素收率为82.95%。
产品中不含有毒的有机溶剂，适合于以营养补充剂和食品添加剂的形式使用。此结晶的应用形式可为油悬液（与植物油混合乳化）、珠粒(通过喷雾冷凝得到的微胶囊)、干粉(通过喷雾干燥得到的微胶囊)等。
实施例 2
500g 万寿菊油树脂(总叶黄素含量14.5%)与250mL丙醇混合，升温至85℃,搅拌直至形成一种均一流动的溶液，边搅拌边缓慢滴加210mL50%KOH溶液，滴加时间为60分钟，在此温度下皂解反应3小时。
反应液降温到60℃,加入乙酸乙酯溶液1500mL，乳化剂Span-40 65g后搅拌0.5hr使其混合均匀。缓慢滴加50%的乙醇钠溶液100mL，滴加时间为1.0hr，同时升温到80℃,搅拌反应6.0hr。
反应完成后加入1000mL去离子水和250mL乙醇，稀释过程中溶液温度保持在60℃，慢速搅拌120分钟。用过滤方法分离所得到的结晶，此分离过程比较容易，30分钟内操作完成。收集到的结晶用85℃热水洗2-3次直至洗出液几乎无色，最后用无水乙醇淋洗，再在40℃下进行真空干燥直至其干燥失重小于5%。
最后得到61.0g成品，其中含85.6%总类胡萝卜素（用紫外-可见光光度仪分析）,这些类胡萝卜素中含15.2%全反式叶黄素，84.8%全反式玉米黄质(用HPLC分析)，其余的为微量其它类类胡萝卜素。总类胡萝卜素收率为72.02%。
产品中不含有毒的有机溶剂，适合于以营养补充剂和食品添加剂的形式使用。此结晶的应用形式可为油悬液（与植物油混合乳化）、珠粒(通过喷雾冷凝得到的微胶囊)、干粉(通过喷雾干燥得到的微胶囊)等。
实施例 3
1000g 万寿菊油树脂(总叶黄素含量14.5%)与2500mL乙醇混合，升温至75℃,搅拌直至形成一种均一流动的溶液，边搅拌边缓慢滴加380mL50%KOH溶液，滴加时间为60分钟，在此温度下皂解反应4小时。
反应液降温到60℃,加入丙醇溶液1200mL，乳化剂Tween-60 60g后搅拌0.5hr使其混合均匀。缓慢滴加37%的甲醇钠溶液100mL，滴加时间为1.0hr，同时升温到80℃,搅拌反应3.0hr。
反应完成后加入1000mL去离子水和250mL乙醇，稀释过程中溶液温度保持在60℃，慢速搅拌120分钟。用压滤方法分离所得到的结晶，此分离过程比较容易，30分钟内操作完成。收集到的结晶用85℃热水洗2-3次直至洗出液几乎无色，最后用无水乙醇淋洗，再在40℃下进行真空干燥直至其干燥失重小于5%。
最后得到141.6g成品，其中含84.7%总类胡萝卜素（用紫外-可见光光度仪分析）,这些类胡萝卜素中含87.1%全反式叶黄素，12.8%全反式玉米黄质(用HPLC分析)，其余的为微量其它类类胡萝卜素。总叶黄素收率为82.71%。
实施例4-7
项目/步骤 实施例4 实施例5 实施例6 实施例7
原料数量(g)和其中叶黄素含量 300g/14.5% 1500g/13.6% 480g/15.4% 600g/14.6%
皂解反应时溶剂种类及数量 甲醇/600ml 乙醇/1200ml 正丙醇/500ml 乙醇/800ml
皂解反应时碱催化剂种类及用量 37%甲醇钠溶液/118ml 50%乙醇钠溶液/816ml 50%KOH乙醇溶液/1780ml 45%NaOH溶液/400ml
异构化反应前补加溶剂种类及数量 乙酸乙酯/300ml 乙醇/1500ml 异丙醇/1200ml 乙酸异丁酯/1800ml
异构化反应前补加乳化剂种类及数量 Tween-60/ 17.5g Span-40/ 204g Tween-80/ 52.0g Span-20/ 88.0g
异构化反应前滴加碱液种类及数量 50%乙醇钠溶液/ 87ml 50%KOH乙醇溶液/820ml 45%NaOH溶液/ 330ml 37%甲醇钠溶液/ 50ml
异构化反应温度 90℃ 60℃ 70℃ 85℃
异构化反应时间 3.0hr 1.0hr 4.5hr 6.0hr
终产品数量(g) 42.9g 178.8g 71.5g 72.5g
终产品中总类胡萝卜素含量(%, UV) 76.4% 82.3% 74.8% 88.4%
终产品中叶黄素和玉米黄质的比例(HPLC) 94.6%/5.2% 85.2%/14.5% 30.4%/69.4% 15.2%/84.7%
比较实施例
1000g 万寿菊油树脂(总叶黄素含量14.5%)按US Patent 7,271,298揭示的工艺条件皂解并通过醇水溶液去除相关杂质后得到类胡萝卜素晶体129.6g，含量为93.2%，产品收率83.3%，其中全反式叶黄素占93.1%，全反式玉米黄质占6.8%。
取上述叶黄素晶体120g按US Patent 7,485,738公开的方法进行差向异构化反应，通过HPLC色谱法监控反应进行程度，1.5hr后停止反应，经再次醇水溶液去除相关杂质，最终得类胡萝卜素晶体71.3g，含量为85.6%，全反式叶黄素比例为88.1%，全反式玉米黄质比例为11.8%，差向异构化反应收率为54.6%。
上述两步反应的总收率只有45.5%，远低于本发明得到的产品收率。

Claims (9)

  1. 一种以植物油树脂为原料制备含较高玉米黄质含量的叶黄素晶体的方法，具体步骤如下：
    a)采用含叶黄素二酯的植物油树脂与食品级醇类溶剂混合，通过搅拌，形成均一的能自由流动的混合液，再在碱性环境下、40-85℃下皂解该混合液3-5小时；
    b)向a)所得混合液补加有机溶剂和加入乳化剂，搅拌均匀；
    c)向b)所得混合液中滴加强碱溶液，使得到的部分叶黄素晶体差向异构化转化成玉米黄质；
    d)用去离子水和醇类溶剂组成的混合溶剂稀释c)所得的反应液，使溶液中溶质的体积浓度降低至10-50%，去离子水与醇类溶剂的用量分别为原料植物油树脂重量的2-10倍与0.5-2倍，然后升温至60-75℃，慢速搅拌0.5-2.0小时，使类胡萝卜素形成颗粒状结晶析出；
    e)采用离心或过滤方法得到d)中生成的结晶；
    f)用60-85℃的热水漂洗e)中得到的结晶2-3次，直至洗出的母液接近无色；
    g)干燥之前结晶用无水乙醇淋洗滤饼一次，然后采用真空干燥或冷冻干燥得到叶黄素晶体，使其最终的干燥失重小于5%。
  2. 根据权利要求1所述的方法，其特征在于植物油树脂选用万寿菊花、金盏花、菠菜、草莓、椰菜、甘蓝或玉米的提取物。
  3. 根据权利要求1或2所述的方法，其特征在于步骤a)中，食品级醇类溶剂的加入量为植物油树脂重量的0.5-2.0倍体积。
  4. 据权利要求3所述的方法，其特征在于用到的强碱为NaOH、KOH、甲醇钠或乙醇钠，皂解时所需的碱量为植物油树脂中叶黄素二酯质量的0.5-6.0倍，异构化反应时，碱的用量为植物油树脂中叶黄素二酯质量的0.1-1.0倍。
  5. 根据权利要求1所述的方法，其特征在于补加的有机溶剂选用醇类溶剂或酯类溶剂，其用量为植物油树脂重量的1.0-3.0倍体积。
  6. 根据权利要求5所述的方法，其特征在于补加的有机溶剂选用乙醇、异丙醇、丙醇、乙酸乙酯或乙酯异丁酯。
  7. 根据权利要求1所述的方法，其特征在于所述的乳化剂选用吐温系列乳化剂或司班系列乳化剂，其用量为植物油树脂中叶黄素晶体质量的0.4-1.0倍。
  8. 根据权利要求1所述的方法，其特征在于异构化反应时，反应时间的选用范围为0.5-6.0hr，温度的选用范围为60-90℃。
  9. 根据权利要求1所述的方法，其特征在于所述的醇类溶剂为甲醇、乙醇、异丙醇或丙醇。
PCT/CN2010/071061 2010-01-07 2010-03-18 一种以植物油树脂为原料制备含较高玉米黄质含量的叶黄素晶体的方法 WO2011082560A1 (zh)

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