CN1272059A - Carotenoid formulations - Google Patents

Carotenoid formulations Download PDF

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CN1272059A
CN1272059A CN98807515A CN98807515A CN1272059A CN 1272059 A CN1272059 A CN 1272059A CN 98807515 A CN98807515 A CN 98807515A CN 98807515 A CN98807515 A CN 98807515A CN 1272059 A CN1272059 A CN 1272059A
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lycopene
compositions
carotenoid
food
concentration
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L·施利帕利刘斯
M·斯特拉汉
D·X·赫莱
E·G·斯佐克
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Henkel AG and Co KGaA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/01—Hydrocarbons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/01—Hydrocarbons
    • A61K31/015—Hydrocarbons carbocyclic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles

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  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
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Abstract

The present invention provides compositions of lycopene dispersed in medium chain triglyceride oil obtained from the esterification of fatty acids and glycerol.

Description

Carotenoid formulation
Technical field of the present invention
The present invention relates to carotenoid compositions or carotenoid is dispersed in by the preparation in the oil of fatty acid and glycerine esterification gained.More particularly, the present invention relates to carotenoid compositions or the carotenoid preparation in medium chain triglyceride oil.
Background technology of the present invention
In recent years, no matter be research worker or enterprise, the research interest of this compounds of being called carotenoid is all being increased.Carotenoid is the natural materials that extensively is present in the plant and animal, and colored compound normally, and they make fruits and vegetables be yellow to red mostly.The Crustaceans for example lacteous of the redness of Lobster and butter also is because due to the carotenoid.Epidemiological study shows that carotenoid lycopene (the main diet source of lycopene is a Fructus Lycopersici esculenti) has antitumaous effect in human body.Yet, up to the present, without any the Research on ability that in the animal model of having set up, suppresses tumor development about consumption of lycopene.
Because carotenoid also has the high strength color at low concentration, and their cost of extraction is very high from natural goods, so in the middle of this century, the organic chemist has successfully synthesized some and had the carotenoid of important economic worth.Beta-carotene and canthaxanthin and some derivants have become commercially available food coloring, food additive and animal pigment precursor, particularly the commodity of being sold by Europeancompanies Hoffman-LaRoche AG and BASF AG.
Carotenoid has unique conjugation carbon-to-carbon double bond chemical constitution.In main commercially available carotenoid, 9 two keys are arranged, they make molecule have special physics and chemical property.Especially, the reactivity of carotenoid and oxygen and free radical is very strong, so carotenoid can be classified as antioxidant commonly used.Carotenoid is present in people, the animal and plant tissue.For the reaction that the emergency rating of environment for example is higher than normal temperature and highlight strength, also can produce the carotenoid of higher concentration in some life entities.The dissolubility of carotenoid in water is very low usually, so it is positioned at the film and the lipid aggregation of living cells.In life entity, the concentration of carotenoid is quite low usually, is approximately 10-100ppm.
In the commercially available carotenoid product of the high concentration process that preparation can extract now and can make by chemical synthesis process, need stabilization of carotenoids so that it is not by airborne oxygen oxidation from natural goods.The carotenoid molecule also can be degraded owing to enzyme or microbial action, but this situation takes place under low concentration (for example being lower than 0.1%) and water environment usually.The chemical constitution of oxidation rate and carotenoid has much relations.The reactivity of some carotenoid is very strong, and the initiation reaction automatically in air of its homodisperse high concentration crystal form.Lycopene is wherein a kind of common carotenoid of the easiest oxidation, and in the process of preparation commercially available prod, stablizes lycopene or contain the compositions of lycopene extremely important for entire product.The oxidation that can not prevent some carotenoid has brought the commercialization difficulty to it.
It is confirmed that stablizing of carotenoid is very difficult usually.Wherein a kind of to help the means of anti-oxidation be to preserve pure crystal class carotene material in noble gas, yet this only is of practical significance on laboratory scale, not commercial value.Also known carotenoid compositions is suspended in the triglyceride oil (for example soybean oil) to help prevent oxidation.Though (carbon chain length is mainly C to use the soybean oil that extracts with solvent and Semen sojae atricolor according to proof 16-C 18Oil) very effective to some carotenoid such as beta-carotene, but it can not stop the degraded of other carotenoid such as lycopene on commercial size.
The invention summary
Therefore, the method that the purpose of this invention is to provide anti-carotenoid oxidation.Surprisingly, have been found that carotenoid compositions or carotenoid are in the oxidation that can alleviate carotenoid by suspension in the oil (not being the oil that extracts with solvent) of fatty acid and glycerine esterification gained or dispersion liquid from natural goods.In addition, find that carotenoid compositions or carotenoid are C in chain length 6-C 12Oil (hereinafter referred to as " medium chain triglyceride oil ") in suspension or the dispersion liquid oxidation that makes carotenoid significantly descend.
Brief description of drawings
Accompanying drawing 1 illustrates the stability (respectively testing the meansigma methods of bottle) of two kinds of compositionss described in the embodiment 1.
Accompanying drawing 2 has been described the timeline of embodiment 2 described 10 weeks tests to determine the maximum tolerated dose (MTD) of lycopene in male rat (colon models) and female rats (breast model).
The HPLC data of accompanying drawing 3 expression echinenones and lycopene.
Accompanying drawing 4 expression all 5 kinds of standard class carotene, i.e. the HPLC data of cryptoxanthin, canthaxanthin, echinenone, lycopene and beta-carotene.
The accompanying drawing 5 expressions standard curve of the lycopene of crystalline lycopene and echinenone generation.
The HPLC analysis result of accompanying drawing 6 expression lycopene suspensions.
The chemical constitution of the carotenoid that exists in the accompanying drawing 7 expression lycopene suspensions.
Accompanying drawing 8-11 represents the influence that lycopene increases the weight of animals.
The HPLC-MS analysis result of the saponification organic extract that accompanying drawing 12 expression is extracted from liver, this result shows and wherein contains a large amount of lycopenes, phytofluene and phytoene and their geometric isomer.
The detailed description of the preferred embodiment of the invention
On the one hand, the invention provides and comprise carotene compositions or carotenoid at the preparation by the suspension/dispersion in the oil of aliphatic acid and glycerine esterification gained. Use the oil of esterification so basically to reduce to occur in the preparation possibility of the impurity of impurity-namely in the oil that from natural goods such as soybean, extracts with solvent, exist. Preferably carry out esterification with basically pure aliphatic acid and glycerine. Preferably come purification of fatty acid by distillation. Used oil is preferably medium chain triglyceride oil. Triglycerides is the ester that aliphatic acid becomes with glycerine. Key component-the triglycerides of fat and oil has general formula CH2(OOCR 1)CH(OOCR 2)CH 2(OOCR 3), R wherein1、R 2And R3Can have different chain length. In context of the present invention, the chain length of preferred triglycerides is C6-C 12, more preferably C8 -C 10。
On the other hand, the invention provides the preparation that comprises carotenoid compositions or the suspension/dispersion of carotenoid in medium chain triglyceride oil.Again preferably, the chain length of medium chain triglyceride oil is C 8-C 10Described medium chain triglyceride oil preferably makes with fatty acid and glycerine esterification.Equally, preferred purified basically fatty acid and the glycerol of using carries out esterification, wherein preferably comes purification of fatty acid by distillation.
In the preferred form of the present invention, carotenoid compositions mainly comprises carotenoid.Carotenoid is preferably selected from lycopene, beta-carotene, sigma carotene and phytofluene or their mixture.Lycopene is preferred carotenoid.Carotenoid compositions and carotenoid preferably derive from natural goods.
In the preferred embodiment of the invention, medium chain triglyceride oil is commercially available Delios V TM, this is the medium chain triglyceride oil of being produced by Grunan Lebensmitteltechnologie.Another preferred medium chain triglyceride oil is registered as CAS 73398-61-5, and this is the mixed triglyceride made from sad and capric acid.This medium chain triglyceride oil can its trade name Neobee From Stepan Co., Northfield, lllinois buys.The acid monomers of medium chain triglyceride oil is preferably selected from C 6Caproic acid, C 8Sad, C 10Capric acid and C 12Lauric acid or their mixture.The preferred fatty acid and the glycerol of purification of using prepares medium chain triglyceride oil, because use these materials can reduce to contain in the oil probability that also can cause the impurity of carotenoid oxidation.In preferred embodiment, medium chain triglyceride oil derives from natural goods.
In another embodiment preferred of the present invention, carotenoid account for weight of formulation up to 10%.Carotenoid preferably accounts for the 4%-5% of weight of formulation.In another embodiment preferred of the present invention, oil account for weight of formulation up to 98%, more preferably 20%-80%, 30%-70% most preferably.
Compositions preferably contains oil-soluble inhibitor.Add antioxidant and further help to prevent the carotenoid oxidation.Antioxidant is preferably selected from tocopherol, fourth hydroxyanisol, butylated hydroxytoluene, propyl gallate, ethoxy quinoline and ascorbyl palmitate and other Natural antioxidant extract that can obtain from medical herbs, preferred natural tocopherol, and composition thereof.Tocopherol be preferably selected from β-, γ-and Delta-Tocopherol or their mixture.The present invention other preferred embodiment in, antioxidant account for weight of formulation up to 15%, preferred 5%-10%.
For oral administration, the present composition also optionally also contains the pharmaceutically suitable carrier that is suitable for oral administration.Orally administered composition of the present invention can make with the known conventional mixed method of pharmaceutical field, is about to active substance and is suitable for being administered systemically and commonly used in peroral dosage form, edible pharmaceutically acceptable nontoxic inertia, solid-state or liquid carrier and/or mixed with excipients.Combination of oral medication can be made tablet, lozenge, chewing gum and the capsule that comprises slow releasing tablet.Perle dosage form most preferably.Those skilled in the art can prepare these dosage forms according to technology known in the art, for example in Remington ' s Pharmaceutical Sciences, the 18th edition (1990), MackPublishing Co., Easton, the technology of describing among the PA.
The method that the present invention also provides biology can utilize antioxidant comprises effective dose suspended or being dispersed in lycopene in the medium chain triglyceride to host's oral administration.In context of the present invention, be to be understood that term " host " not only comprises the people, also comprises non-human animal, especially mammal.This method comprises, with the compositions of the oil suspension/dispersion liquid that comprises carotenoid of effective dose, for example with the dosed administration of about 1-1000 μ g/kg carotenoid in patient's body.Preferred dosage is about 10-100 μ g/kg carotenoid.In implementing the inventive method process, but the present composition is administered once every day or administration every day repeatedly.Therapeutic scheme may be during prolonging in administration.Each dosage or total dosage will be determined the factors such as toleration of this chemical compound according to the type of for example disease and the order of severity, patient's age and health status and patient by the doctor.
The present invention also provides and treated or prevented method for cancer in the patient of needs treatment.In context of the present invention, be to be understood that term " patient " not only comprises the people, also comprises non-human animal, especially mammal.This method comprises, with the compositions of the oil suspension/dispersion liquid that comprises carotenoid of effective dose, for example with the dosed administration of about 1-1000 μ g/kg carotenoid in patient's body.Preferred dosage is about 10-100 μ g/kg carotenoid.In implementing the inventive method process, but the present composition is administered once every day or administration every day repeatedly.Therapeutic scheme may be during prolonging in administration.Each dosage or total dosage will be determined the factors such as toleration of this chemical compound according to the type of for example disease and the order of severity, patient's age and health status and patient by the doctor.
Can by with the present composition itself or with its with other active component that comprises other antioxidant and/or therapeutic agent administration in pharmaceutical composition implement this method.Other is applicable to that but therapeutic agent of the present invention is the arbitrary compatibe drug that can realize the object of the invention by identical or other mechanism effectively, or its active activity with preparation of the present invention or compositions has additional or synergistic medicine.
Employed chemical compound of therapeutic alliance or activating agent can with present composition administration simultaneously in different or same preparation, or, so just reached the effect of therapeutic alliance in different time administration, for example administration successively.Dosage and scheme will be regulated by the doctor, preferred its standard dose that reduces earlier, and definite then result who is obtained determines best dosage and scheme.
In the detailed description of the preferred embodiment of the invention and the back claims, further explained feature of the present invention below, it is clearer that above and other objects of the present invention, advantage and feature will become.
Embodiment
Embodiment 1: preparation and stability test
For the improvement that confirms esterised oil antioxidation, carry out two comparative tests with containing the carotenoid compositions (" lycopene compositions ") of lycopene as carotenoid.One of them test comprises the suspension/dispersion of lycopene compositions in the soybean oil that extracts with solvent from Semen sojae atricolor, another test comprises that the lycopene compositions is by the suspension/dispersion in the medium chain triglyceride oil of fatty acid and glycerine esterification gained.Comparative test result then.
Material
Testing used lycopene compositions extracts from Fructus Lycopersici esculenti.
Used medium chain triglyceride oil is by the fatty acid fractional distillation with Oleum Cocois, then selected fraction and high-purity glycerine esterification with fatty acid of 8 and 10 carbon atoms is made.Form oil by this way and can reduce the impurity that in the oil that directly from animals and plants, extracts, is occurred.Soybean oil is the standard oil that extracts from Semen sojae atricolor.
Two batches of 10 kilograms of lycopene compositionss are dispersed in respectively in each 15 kilograms of soybean oil (" compositions 1 ") and medium chain triglyceride oil (" compositions 2 ") as diluent or suspension/dispersed oil, it are mixed forming continuous phase with effective mixed method.The sample of respectively getting 1 each compositions of gram is placed in the vial with test.
Test process and result
Measure the concentration of lycopene in compositions 1 and the compositions 2 respectively with spectrophotometer, to determine concentration at following time lycopene:
0 month (be after the preparation of compositions soon);
1 month;
2 months; With
3 months.
(a) analytical method
Each sample is dissolved in the chloroform, and is diluted to debita spissitudo with cyclohexane extraction.At 20 ℃, certain wave strong point mensuration absorbance, by known extinction coefficient calculating concentration.
Use following reagent and instrument:
The spectrophotometer of being furnished with the 10mm glass cell
Analytical balance
The 100ml volumetric flask
The 50ml volumetric flask
2.0ml bulb pipette
Chloroform, the AR level
Cyclohexane extraction
When testing, the medicine lycopene is slowly degraded under illumination, therefore uses low flashed glass.For each sample, all carry out following operation.The lycopene oil suspension of each 0.5-1.0 gram 5% is weighed accurately to respectively in the 100ml volumetric flask.Add about 5ml chloroform, reagent is fully mixed so that the sample dissolving.
Observe each sample facing to light, dissolve fully to guarantee it.
Dissolve fully in order to ensure each sample, it was placed 5 minutes, add the 5ml chloroform again, and insulation (if necessary) under hot tap-water.
With cyclohexane extraction solution dilution to volumetric flask volume is also fully mixed.This is a solution A.The suction of 2ml solution A is measured in the 50ml volumetric flask, be diluted to volumetric flask volume and fully mixing with cyclohexane extraction.This is a solution B.
The suction of 2ml solution B is measured in the 50ml volumetric flask, be diluted to volumetric flask volume and fully mixing with cyclohexane extraction; This is a solution C.
Spectrophotometer (having the 2nm slit width) being returned to zero in two cells with cyclohexane extraction, is blank with the cyclohexane give, measures the absorbance of solution C at 472nm wavelength place.This analysis repeats twice.
In the result of calculation process, use following standard:
The A=solution C is at the absorbance at 472nm wavelength place
The m=sample weight
E=is at the extinction coefficient E1%/1cm=3450 at 472nm wavelength place
The concentration of C=lycopene (%)
C=(A * dilution factor)/(E * m)
In the interval of test, two kinds of compositionss are all stored under 25 ℃ of Celsius temperatures.
Find that at test period lycopene is tending towards forming megacryst after the storage several months, crystal can reach 2-3mm sometimes.Because the megacryst that forms may have a strong impact on analysis result owing to error of sampling,, and under 25 ℃ of temperature, store so this sample of material of the gram of 1 in the vial is the sample of taking immediately after the preparation.Every interval one month is got bottle and is detected, and detects whole content and overcome the problem of sampling that crystallization brings.
(b) result
Shown in accompanying drawing 1 and table 1, that the stability test result of two kinds of compositionss in each test bottle is average:
Table 1
Compositions 1: lycopene is in soybean oil Under 25 ℃ of temperature, store
The preparation back time 0 month 1 month 2 months 3 months
The content % (weight ratio) of lycopene ???5.0 ??4.3 ??3.5 ??2.9
Compositions 2: lycopene is in medium chain triglyceride oil Under 25 ℃ of temperature, store
The preparation back time 0 month 1 month 2 months 3 months
The content % (weight ratio) of lycopene ???4.3 ??3.9 ??4.4 ??4.3
Discussion of results
Above-mentioned result of the test shows, during 3 months in, the constant concentration of lycopene in medium chain triglyceride oil (compositions 2), in fact, average percentage concentration is identical with instant concentration after compositions 2 prepares after 3 months.During 3 months, lycopene is the concentration determination result in soybean oil (compositions 1) show as, and the % weight concentration of lycopene descends steadily.
These results confirm, have the Stabilization that prevents the lycopene oxidation by the oil (being medium chain triglyceride oil in this test) of fatty acid and glycerine esterification gained.
Embodiment 2: the clinical preceding toxicity and the stability of lycopene
The purpose of present embodiment test is that the biology of determining lycopene in the food can receive dosage range, picked-up and tissue distribution.
Specific purposes
The lycopene of determining administration in AIN-76A food in male and female Fischer 344 rats maximum tolerated dose and estimate picked-up and the distribution of lycopene in blood, feces and Different Organs such as mammary gland, prostate, colon and lung in the food.
Method
Inbred male (n=70) and female (n=70) Fischer 344 rats derive from Taconic Farms, Germantown, and NY. is long by about 35 days big hour rat, and all rats are transferred to the laboratory from isolation area.With conventional randomized rat is distributed into the test group then, comprises rat with identical average weight to guarantee each group.
Shown in table 2 and accompanying drawing 2, carry out the test of 10 weeks, measure the maximum tolerated dose (MTD) of lycopene in male rat (colon models) and female rats (breast model).The selection of dosage range is the value according to disclosed beta-carotene.The blended carotenoid compositions that will contain 5.7% carotenoid, wherein comprises 3.7% pure lycopene is suspended in (said composition is called " lycopene suspension " hereinafter) in the medium chain triglyceride.This lycopene suspension is mixed in the AIN-76A food.
Table 2:MTD test
The experimental animal number The amount of lycopene a
Organize male female (mM/kg food) estimates (% food) (mM/kg food) (ppm)
1 10 10 2 10 10 3 10 10 4 10 10 5 10 10 6 20 20 total numbers 70 70 2.5????????????1280???????????0.128 1.0????????????512????????????0.0512 0.5????????????256????????????0.0256 0.25???????????128????????????0.0128 0.10???????????51?????????????0.0051 0??????????????0??????????????0
aBased on lycopene MW=536
Under the low light level, prepare food, and be stored in-4 ℃ of sealed containers under the temperature with 4 kilograms of batches.The composition of AIN-76A food is as shown in table 3.
Table 3: standard A IN-76A food
Component content (g/100g)
Junket egg 20 cornstarch 52 glucose 13 corns 5 DL-egg ammonia 0.3 choline bitartrate 0.2 Alphacel 5 AIN-76 vitamin mixtures 1.0 AIN-76 machine thing mixtures 3.5
Total amount 100
Energy value (kilocalorie/g) 3.89
By in diet, replenishing the Neobee of appropriate amount (Stepan Co., Maywood NJ) control the difference that lipid that each treatment group absorbed contains to medium chain triglyceride, comprise that like this all rats of matched group have just absorbed the edible fat of same amount.
Before the off-test, 3 rats in each group are placed metabolic cage, gather urine in 24 hours and feces to carry out lycopene analysis subsequently.Gather blood by the cardiocentesis under narcotism and carry out the lycopene analysis, pass through CO 2All rats are put to death in anesthesia.At-20 ℃ of stored bloods and feces.When necropsy, all major organs systems are carried out Rough Inspection.The tissue of arbitrary display abnormality is placed formalin, and preserve to carry out later histological examination.
Write down rat body weight weekly one time, estimate rat appearance (barment tag) and food elusive behavior.By concentrating, maximum tolerated dose is the dosage that causes average weight decline＞10%.
Table 4: the HPLC method that detects lycopene
Pump Waters Model 510
Detector Shimadzu, SPD-10A UV-Vis detector
Integrator Waters 745 Data Model
The automatic gradient controller of controller Waters
Post Rainin Microsorb 5 μ mC18,25cm * 4.6mm
Wavelength 470nm
Speed 0.7ml/ minute
AUFS?????0.032
Drawing speed 0.25cm/ minute
Sample size 20-50 μ l
Solvent orange 2 A 90% acetonitrile 10% methanol
Solvent B
45% hexane 45% dichloromethane 10% methanol
Every 500ml solvent B adds the 0.5ml diisopropylethylamine
Program
Time solvent orange 2 A % solvent B% 0 95 5
?????10?????????95????????????5
?????40?????????55????????????45
?????45?????????95????????????5
?????65?????????95????????????5
The sample introduction solvent mixture
40% acetonitrile
20% methanol
20% hexane
20% dichloromethane
By HPLC quantitative assay lycopene
HPLC device, solvent system and solvent programming are listed in the table 4., comprising as standard with the carotenoid crystallization: alpha-carotene, beta-carotene, lycopene, cryptoxanthin, kryptoxanthin and echinenone (Hoffmann-LaRoche).The ketone derivatives echinenone that uses beta-carotene is as interior mark.With sample introduction solvent (table 4) reference material is prepared into the stock solution that concentration is 30 μ g/ml.
(1) lycopene (RO 01-9251-00)
(2) cryptoxanthin (RO 01-9371-000)
(3) kryptoxanthin (RO 04-0763-000)
(4) 9-carotene (RO 01-8300-000)
(5) echinenone (RO 04-2847-000)
By with 5 days time ratios from the variability and the reliability (table 5) of this system of various original detection of the standard solution sampling of lycopene and echinenone.The retention time of lycopene changed to the highest 35.95 minutes from minimum 32.58 minutes; The retention time of echinenone changed to the highest 30.64 minutes from minimum 28.54 minutes.The ratio of lycopene/echinenone changes to the highest by 1.20 from minimum 1.12.When comparison curves lower integral area, found difference (table 5) in the similar low-level sample.
By using crystal of lycopene and echinenone, produced the standard curve (accompanying drawing 5) of lycopene.Test value reads from standard curve, and represents with ng/ml or ng/ μ g/g tissue wet.
Table 5: carotenoid HPLC test: variability and reliability
Test program RT lycopene
Lycopene echinenone ratio (lycopene/ratio (lycopene/
Echinenone) lycopene echinenone echinenone)
First day
1?????????33.17????29.15??????1.14???????????586,132????720,121??????0.81
2?????????32.91????2?8.89?????1.14???????????567,339????755,926??????0.75
3?????????33.49????29.86??????1.12???????????527,365????737,088??????0.72
4?????????34.48????30.47??????1.13???????????532,491????765,646??????0.70
5?????????34.47????30.45??????1.13???????????527,024????772,265??????0.68
6?????????34.64????30.64??????1.13???????????496,268????768,335??????0.65
The 3rd day
7?????????34.05????28.30??????1.20???????????511,111????781,883??????0.65
8?????????33.04????29.03??????1.14???????????551,826????736,851??????0.75
The 5th day
9?????????32.99????28.78??????1.15???????????624,986????803,354??????0.78
10????????32.58????28.32??????1.15???????????573,498????793,606??????0.72
11????????32.81????28.54??????1.15???????????562,057????823,087??????0.68
12????????35.95????30.11??????1.19???????????612,266????859,039??????0.71
13????????34.50????29.86??????1.16???????????543,684????869,292??????0.63
X?????????33.78????29.42??????1.15???????????555,085????783,576??????0.71
S?????????0.99?????0.85???????0.023??????????37,924?????45,566???????0.053
C.V. (%) 2.9 2.9 2.0 6.8 5.8 7.5RT=retention times (minute) integral area under the AREA=curve
Food
The lycopene that extracts from food as described below: the sample that will contain lycopene is suspended in the sample introduction solvent (table 4), and homogenate in multitube (polytron) homogenizer.Add echinenone as interior mark, with mixture under 4 ℃, 3000rpm rotating speed centrifugal 15 minutes.Inclining supernatant, preserves, and repeats to extract.Then with the extract that merges under nitrogen in 60 ℃ of dryings, residue is dissolved in the 0.5ml sample introduction solvent, carry out HPLC then and analyze.Same method is carried out minimum variation, is applicable to feces and lung tissue.The efficient of extracting lycopene is shown in table 6A and B.Under all test concentrations, extraction efficiency is not from 75% 100% waiting under the minimum lycopene concentration of maximum concentration.
Serum
The extraction lycopene is performed such from serum: 1ml serum is added in 0.1ml echinenone reference material and the alcoholic acid mixture of 0.9ml.Mixture vortex vibration 20 seconds, add the 2.0ml hexane then, acutely mixed 1 minute.Then that this is muddy mixture under 4 ℃, 3000rpm rotating speed centrifugal 15 minutes, inclining clarifying hexane layer.Above-mentioned steps is repeated 3-5 time.Merge the hexane part, then under nitrogen in 60 ℃ of dryings, residue is dissolved in the 0.5ml HPLC solvent, carry out HPLC and analyze.
Tissue
Breast, liver and prostata tissue contain the lipid that can disturb this analysis.Therefore, the saponification step of tissue need be carried out before organ extracts.To be organized in and add echinenone as homogenate in the interior target 20ml methanol.For the breast tissue that comprises 85% adipose cell, add the NaOH of 8.0ml 50% and the sodium ascorbate of 4.0ml 25%, with mixture homogenate.For prostate that contains less lipid and liver organization, add the NaOH of 4.0ml 50% and the sodium ascorbate of 2.0ml 25%, with mixture homogenate.Saponification is in the 50ml pipe saturated with nitrogen atmosphere overnight, carries out in 30 ℃ in temperature control electromagnetic shaker is bathed.After the saponification, in each pipe, add the 10ml hexane, vortex vibration 1 minute.With mixture under 4 ℃, 2000rpm rotating speed centrifugal 10 minutes, inclining hexane layer and preserves.This hexane extraction step is repeated 5-7 time.Merge hexane extract, then under nitrogen in 60 ℃ of dryings, residue is dissolved in the sample introduction solvent, carry out HPLC and analyze.Except not carrying out saponification, from lung and colon, extract lycopene by same way as.
Table 6A﹠amp; B: the efficient of from AIN-76A food, extracting lycopene
6A
The group number lycopene
Concentration extract sample size in food extracts the concentration of back lycopene
Sample weight
Volume
(μg/g)??????(g)??????(ml)????????(μl)????????????(μg/g)
1???????????????1,280?????0.5??????100?????????20??????975?????(76％)
2???????????????512???????0.5??????100?????????20??????321?????(63％)X
403?????(79％)
3???????????????256???????0.5??????50??????????20??????192?????(75％)
4???????????????128???????0.5??????50??????????50??????122?????(95％)X
101?????(86％)
5???????????????51????????1.0??????50??????????50??????62??????(100％)X
52??????(100％)
6B
Numbering echinenone lycopene ratio (lycopene/concentration
Echinenone) (μ g/g)
Area under curve
Standard 2,590,806 2,191,332 0.85
2,754,253????2,505,114????0.91
1????????????2,891,211????4,493,499????1.55?????????975
2????????????2,980,758????1,527,316????0.51?????????321X
2,618,266????1,663,187????0.64?????????403
3????????????2,804,003????1,705,160????0.61?????????192
4????????????2,236,710????2,179,322????0.97?????????122X
2,636,044????2,087,983????0.80?????????101
5????????????2,647,478????2,620,900????0.99?????????62X
2,613,385????2,171,047????0.83?????????52
I. the analysis of lycopene suspension
As lycopene suspension (Henkel Corporation, La Grange, IL) be by organic extraction method extract the time, main carotenoid is lycopene, but also has many other known carotenoid that exist in Fructus Lycopersici esculenti, α-and beta-carotene, phytofluene and phytoene (accompanying drawing 6 and 7).With the above-mentioned HPLC system that photodiode array detector is installed the carotenoid in the lycopene suspension is carried out quantitative analysis.6 kinds of key components are as shown in table 7.
Table 7: the carotenoid quantitative analysis of lycopene suspension
μ g/gm % (g/100g) accounts for the % of carotenoid total amount
1. lycopene 37,504 3.7504 66
2. beta-carotene 12,443 1.2443 22
3. phytofluene 3,349 0.3349 6
4. phytoene 2,794 0.2794 5
5. sigma carotene 440 0.0440 0.7
6. 2,6-encircles lycopene-1,5-glycol 471 0.0440 0.7
Carotenoid total amount 57,001 5.7 100
The content of lycopene is 3.7%, yet the total content of carotenoid is about 5.7%.Therefore, when representing with the % that accounts for the carotenoid total amount, lycopene accounts for 67%, and beta-carotene accounts for 20%; Precursor-the phytofluene of lycopene and phytoene account for 5% respectively, sigma carotene and 2, and 6-encircles lycopene-1, and the 5-glycol accounts for 0.7% respectively.Carotenoid total content according to 5.72% (rather than 4%) recomputates the concentration of lycopene in food, and the result has obtained the concentration of following each treatment group: group 1,1280; Group 2,512; Group 3,256; Group 4,128; With group 5,51 (mg/kg foods).Therefore, the lycopene suspension is not pure lycopene formulations, this suspension contain 5.9% lycopene, α-and beta-carotene, phytofluene and phytoene (precursor of lycopene) be suspended in the suspension in the medium chain triglyceride.Study this for chemoprophylaxis and be actually favourable, because composition that this carotenoid composition of lycopene suspension and content approach carotenoid in the commercially available Fructus Lycopersici esculenti and content are (referring to Khachik, F, Beecher, G.R. dietary factor international conference (International Conference on Food Factors): chemistry and cancer prevention.With the criterion of the distribution of carotenoid in fruits and vegetables as the selection chemopreventive agent.H.Ohigashi (ed.), Springer-Verlag, Tokyo, 1996 (printings)).
II. the stability in food
Once the lycopene suspension is incorporated in the food, just measure its stability with two kinds of methods at once.First method is the food that contains this suspension to be placed for 3 weeks (show 8A, B) in 4 ℃ dark.
Table 8A ﹠amp; 8B: the stability test of lycopene in the food a
8A
Group number date # natural law (in refrigerator) lycopene concentration (μ g/g) % b
1??????5/1???????1????????????????????748??????????????101
2??????5/8???????8????????????????????728??????????????98
3??????5/14??????14???????????????????672??????????????90
4??????5/21??????21???????????????????534??????????????72
aAIN-76A food
bWith the concentration (04/40) of food, 744 μ g/g are as 100%8B group number echinenone aLycopene ratio (lycopene/echinenone) concentration (μ g/g)
TG-AUC standard 2,461,649 2,117,904 0.861 2,304,361 2,486,290 1.08 7,482 2,422,134 2,545,973 1.05 7,283 2,325,379 2,265,087 0.97 6,724 2,334,947 1,787,656 0.77 534
aInterior mark
At different time, take food sample, and extraction lycopene as described below: will about 0.5-1.0g sample be added in the extraction solvent that 15ml is made up of the dichloromethane of 40% acetonitrile, 20% hexane, 20% methanol and 20%.With mixture supersound process 2 minutes, placed then 5 minutes.Incline little red-orange clarified supernatant.Should operate and repeat 4 times, the supernatant that obtains in repetitive process is colourless.Add solvent then to 25-100ml, mixture is centrifugal, and inclining supernatant.Then all supernatant are mixed.Above-mentioned solvent plays two effects: at first, it is the most effective mixture for extracting lycopene, and secondly, it is and the solvent phase solvent together that is used for the lycopene sample is expelled to HPLC.As shown in table 8, after 3 weeks of placement, lycopene is degraded to being about 72% of the 1st day content in 4 ℃ dark; Degraded 10% approximately after 2 weeks.This shows that for this feed test, the period of storage in 2 weeks will be best.
Second method is that the food exposure that will contain lycopene according to this mode, is carried out actual feed test in air and light.Lycopene stable as shown in table 9 in being exposed to the food cup of air and light.
Lycopene stability at room temperature in the table 9:AIN-76A food
The natural law concentration (μ g/g) that date places in the food cup
4/30????????????0???????????????744
5/2?????????????2???????????????702
5/3?????????????3???????????????616
5/6?????????????6???????????????547
5/7?????????????7???????????????439
5/9?????????????9???????????????624
5/10????????????10??????????????576
5/13????????????13??????????????449
5/14????????????14??????????????419
5/15????????????15??????????????425
5/17????????????17??????????????431
5/20????????????20??????????????385
5/21????????????21??????????????376
Obviously, in 48 hours, lycopene only has a small amount of minimizing.After 72 hours, lycopene has reduced 20% approximately.After 3 weeks, the lycopene of primary quantity existing half detect less than.According to these results, food placed in the food cup be no more than 48 hours.
The food that contains the lycopene suspension after 7 months, is measured all main carotenoid (table 10) that wherein exist 20 ℃ of storages.Found that the main carotenoid of all that exist in the lycopene suspension is present in the food with identical relative scale with tomato metabolite of carotenoid.Yet after 7 months, the net loss of the lycopene of processed group 1-5 is respectively 66%, 38%, 40%, 44% and 60%; And in the identical time, compare with initial value, beta-carotene concentration has only descended 10%.
Table 10: with the carotenoid content in the additional food of lycopene suspension a
μg/gm(ppm)
Food mM/kg β-ξ-phytofluene phytoene 2,6-
Food lycopene carotene Radix Dauci Sativae prime ring lycopene
-1, the 5-glycol
1??????2.5?????472????????284??????????1?????????58????????????70????????????2
2??????1.0?????348????????138??????????9?????????28????????????23????????????1.4
3??????0.5?????168????????69???????????4?????????14????????????0.32??????????0.76
4??????0.25????92?????????37???????????2?????????8?????????????5?????????????0.46
5??????0.10????22?????????8????????????0?????????2?????????????1?????????????0.14
6000000 0a foods were stored 7 months at-20 ℃
III. pre-feed test
With carotenoid content is that the food of 2.5mM lycopene (+other carotenoid)/kg food fed for 3 weeks for a male rat.Under this high dose, do not find that body weight or food consumption quantity have any variation.Feces presents significant redness, and the part crust also presents redness.Afterbody is brown.Find that when necropsy liver is peony, caecum and small intestinal also are like this.In stomach fat tissue or prostate and folliculus, found the variable color of little degree.
Lycopene analysis (table 11) in 24 hours in the feces group shows that lycopene nearly 55% total in the food of being taken food is secreted in the feces.This shows that remaining lycopene is absorbed in the intestinal.Also must consider the probability of degrading in the storage process of some lycopenes before extraction.Discovery is rolled into a ball lyophilization with feces before extracting lycopene be the most effectual way of extracting lycopene.This undoubtedly is because exist water and feces group to have geometry in the feces group.Once lyophilization, feces group is powdered easily, and this can more easily enter in the feces basic unit solvent.
Table 11: the analysis of lycopene in the feces
Excretion % efficient in 24 hours feces during food consumption quantity lycopene in the food voided excreta in 24 hours
The estimation of lycopene stool weight content of lycopene lycopene total amount food-intake always
(μg/g)??????(g/24h)???????(μg)????????(g)????????(μg/g)?????(μg)
Organic extraction 744 13-15 9,784 1.44 2,992 4,309 4.309 44
9,784
Lyophilization+
744??????????13-15????????9,784????????1.44????????3,754?????5,406?????????5.405??????55
Organic extraction
9,784
IV. feed test
(1) weight increase
The influence that lycopene increases the weight of animals is shown in table 12 and 13, Fig. 8-11.When estimating by analysis retest difference, be supplemented with between the food rat feeding of various dose lycopene in usefulness, weight increase is without any difference.At 10 weekends, the weight increase degree of control rats than lycopene supplementation group rat height a bit, but this species diversity is no more than 10%, general cut-off point represents that food avoids or toxicity.
Table 12: the lycopene in the food is to female F344 rat aThe influence of weight increase
Group lycopene mixture (ppm) body weight (g * SD)
0 week 8 of week
1??????????????????1280?????86±5??168±10
2??????????????????521??????86±5??169±9
3??????????????????251??????86±5??171±13
4??????????????????128??????86±5??173±8
5??????????????????51???????86±5??168±13
6 contrasts 86 ± 5 172 ± 15
aThe N=10/ group
Table 13: the lycopene in the food is to male F344 rat aThe influence of weight increase
Group lycopene mixture (ppm) body weight (gms * SD)
0 week 8 of week
1???????????????????1280?????113±6??276±30
2???????????????????512??????113±6??280±19
3???????????????????256??????113±6??279±26
4???????????????????128??????113±6??279±12
5???????????????????51???????113±6??280±22
6 contrasts 113 ± 6 285 ± 21
aThe N=10/ group
(2) tissue and serum picked-up
Liver
Compare with serum or other organ, lycopene is wanted dense 100-1000 doubly in liver.Under maximum dose level lycopene (1280ppm), lycopene levels is a 33-120 μ g/g weight in wet base (table 14) in the liver.
Table 14: with being supplemented with lycopene suspension a's
Carotenoid content in the liver of food rat feeding
Female rats
μg/gm?(ppm)
MM/kg 2, and 6-encircles lycopene
ξ-carotene phytoene
Food food lycopene beta-carotene phytofluene-1, the 5-glycol
1??????2.5????120???????????11????????17???????????106????????66?????????9
2??????1.0????64????????????6?????????8????????????48?????????35?????????5
3??????0.5????66????????????8?????????7????????????50?????????40?????????7
4??????0.25???49????????????7?????????2????????????46?????????46?????????4
5??????0.10???42????????????4?????????4????????????33?????????38?????????6
6??????0??????0?????????????0?????????0????????????0??????????0??????????0
Male rat
1??????2.5????33????????????3?????????5????????????30?????????23?????????4
2??????1.0????5?????????????1?????????1????????????5??????????4??????????.39
3??????0.5????60????????????5?????????5????????????84?????????21?????????4
4??????0.25???3?????????????1?????????1????????????-??????????-??????????.28
5??????0.10???12????????????1?????????1????????????10?????????10?????????.34
6??????0??????0?????????????0?????????0????????????0???????????0?????????0
Measure the saponification organic extract of liver by HPLC-MS, the result shows and wherein has a large amount of lycopenes, phytofluene and phytoene and geometric isomer (accompanying drawing 12) thereof.In addition, also detect low-level sigma carotene, all Trans-Beta-Carotenes, 9-suitable-beta-carotene and 13-be suitable-beta-carotene.
In liver samples also detection by quantitative to two kinds of oxidative metabolites of detected lycopene in the human serum-promptly 2 in front, 6-encircles lycopene-1,5-glycol I and II and in human serum undetected its epoxide precursors 2,6-encircles lycopene-1,5-epoxide I and II, detect aforementioned metabolite and show, 2,6-encircles lycopene-1, the 5-glycol is except originating from food, and liver also can be metabolized to lycopene these metabolite.The amount of lycopene shows non-linear dose-response curve, but does not then have in female generally than male height in the female rats liver in male rat.These results show that the carotenoid in the lycopene suspension is absorbed and is stored in the liver, the mode that lycopene is found to be similar in people metabolism in rat.
Serum
The serum lycopene levels is not wait from 80 to 370ng/ml.In the male and female rats of feeding with the food that is supplemented with the lycopene suspension, concentration and the dosage of lycopene in serum is non-linear relation (table 15).Surprisingly, in female rats, in group 3 (200ppm) and group 4 (100ppm) rather than desired group 1 (1000ppm), found maximum concentration.Similar is that in male rat, maximum concentration is in group 3.In female rats, low lycopene (40ppm) group shows minimum serum lycopene concentration, but in male rat, does not then have difference between the highest and minimum lycopene group.
Table 15: the concentration of lycopene (ng/ml) in the rat blood serum a
Group # x ± SD median value range
Female rats
1??????????187±043????205?????123--232
2??????????169±042????180?????109--211
3??????????245±083????210?????174--366
4??????????313±047????308?????262--369
5??????????145±053????152?????081--207
6??????????0???????????0???????0
Male rat
1??????????168±036????160?????134--230
2??????????227±068????225?????148--326
3??????????278±066????285?????174--372
4??????????231±064????215?????153--328
5??????????171±049????177?????100--238
6??????????0???????????0???????0
a??N＝6
These results show that the serum lycopene levels is regulated by Homeostatic mechanism, and this mechanism comprises liver storage mechanism and discharges in the mode that is similar to vitamin A.The serum lycopene levels is also in the people's of the Fructus Lycopersici esculenti of the edible normal level of being reported or tomato products scope in the rat mentioned herein, i.e. 0.1-5 μ g/ml serum (100-5000ng/ml).
Mammary gland
Lycopene concentration in the female rats breast buccal pad is not wait (table 16) from 139 to 460ng/g weight in wet bases.Different with serum, finding has conventional dose-response effect (table 4) between breast tissue and diet lycopene intake.
Table 16: the content of lycopene in the rat mammary gland (ng/g) a
Group # x ± SD median value range
1??????????309±131????235????232--460
2??????????200±030????197????172--231
3??????????215±062????220????139--282
4??????????229±054????217????181--288
5??????????174±057????143????139--239
6??????????0???????????0??????0
1?N＝3
Prostate
Lycopene concentration in the male rat prostate is not wait (table 17) from 32 to 147ng/g weight in wet bases.The mean concentration of lycopene in prostata tissue is than the low order of magnitude of the concentration in breast tissue.The dosage associative mode of picked-up clearly (table 17) in prostate.
Table 17: the content of lycopene in the rat prostate (ng/g) a
Group # x ± SD median value range
1??????????97±17????99????79-112
2??????????95±48????83????54--147
3??????????50±37????35????23--93
4??????????52±26????52????26--77
5??????????47±16????47????32--63
6??????????0
a?N＝3
Lung
Lycopene concentration in the male and female rats lung is not wait (table 18 and 19) from 124 to 424ng/g weight in wet bases.In female rats, between group 1-4, high top effect is arranged, minimum group (group 5) clear and definite reduction is arranged then.In male rat, obtained the analog result except not organizing the increase in 4.
Table 18: the lycopene concentration in the male rat lung (ng/g) a
Lycopene in the food (ngs/gm)
The group # rat is numbered single concentration
The concentration mean concentration
1????????1280??????????????????36-1????????????193
190
36-2????????????187
2????????512???????????????????41-1????????????170
214
41-2????????????257
3????????256???????????????????46-1????????????325
375
46-2????????????424
4????????128???????????????????51-1????????????239
201
51-2????????????162
5????????51????????????????????56-1????????????17
151
56-2????????????135
6????????0?????????????????????61-1????????????0
0
61-2????????????0
Table 19: the lycopene concentration in the female rats lung (ng/g) a
Lycopene in the thing (ngs/gm)
The group # rat is numbered single concentration
The concentration mean concentration
1?????????????1280????????1-1????????????184
227
1-2????????????270
2?????????????512?????????6-1????????????280
246
6-2????????????211
3?????????????256?????????11-1???????????243
193
11-2???????????142
4?????????????128?????????16-1???????????208
211
16-2???????????214
5?????????????51??????????21-1???????????144
134
21-2???????????124
6?????????????0???????????26-1???????????0
0
26-2???????????0
Colon
For colon, can not obtain exact value, because carotenoid enters the little gap of mucosa, this makes can not estimate (absorption) carotenoid and intracavity (unabsorbed) carotenoid in the cell.The colon value is general similar to the value of lung and breast.
Glutathion is analyzed
The principle of measuring glutathion is that the reduction form of these three peptide thiols-glutathion (Glu-Cys-Gly) is participated in several important cell internal reactions directly, comprises the protective effect that prevents free radical damage.Fat carotenoid mutually is different with being arranged in, glutathion is positioned at the aqueous phase of cell, clearly, the antioxidant of fat phase and aqueous phase can interact each other in the following manner: the consumption of other antioxidant can " be saved " or limit to high-caliber antioxidant.In analyzing blood, liver and the kidney all (reduction and oxidation) glutathion (table 20-22).
Table 20: glutathion measurement result-summary in the whole blood
Glutathion (μ mol/g) *Lycopene (ppm) female rats male rat
0????????????1.010±0.0627?????0.936±0.0888
51???????????0.832±0.158 +????1.030±0.0387
128??????????0.890±0.202??????0.920±0.169
256??????????0.848±0.0988?????0.910±0.106
512??????????0.930±0.123??????0.990±0.141
1,280 1.080 ± 0.104 0.948 ± 0.294 *Numerical value is meansigma methods ± S.D., n=5 +With the significant difference of contrast, p＜0.05
Table 21: glutathion measurement result-summary in the liver
Glutathion (μ mol/g) *Lycopene (ppm) female rats male rat
0?????????????5.45±0.366?????6.13±0.516
51????????????6.01±0.309?????6.77±1.53
128???????????6.54±1.09 +????5.38±0.363
256???????????4.87±1.14??????4.59±1.37
512???????????5.88±0.492?????6.90±1.098
1,280 7.38 ± 0.705 +7.16 ± 1.95 + *Numerical value is meansigma methods ± S.D., n=5 +With the significant difference of contrast, p＜0.05
Table 22: glutathion measurement result-summary in the kidney
Glutathion (μ mol/g) *Lycopene (ppm) female rats male rat
0?????????????0.774±0.154??????0.592±0.234
51????????????0.900±0.101??????0.688±0.316
128???????????0.950±0.0758?????0.686±0.170
256???????????1.030±0.414 +????0.624±0.104
512???????????1.090±0.163 +????0.854±0.055 +
1,280 0.089 ± 0.045 +0.828 ± 0.128 + *Numerical value is meansigma methods ± S.D., n=5 +With the significant difference of contrast, p＜0.05
The mensuration of glutathion be by the HPLC with dual Electrochemical Detection carry out (referring to K1einman, W.A.﹠amp; Richie, J.P., " chromatograph magazine " be B672:73-80 (J.Chromatogr), and 1995).The remarkable increase of glutathione level is only arranged, and this situation occurs over just in the liver under high lycopene levels situation.In liver, kidney or whole blood, glutathion does not show the variation relevant with dosage.
Vitamin A and vitamin E analysis in the liver
It is the absorption of retinol and alpha-tocopherol and the influence of storage to two kinds of important fatsoluble vitamiies that research replenishes tomato carotenoids.Use the HPLC method of having set up, find that the concentration of the content of lycopene and retinol and alpha-tocopherol all has very strong linear dose-response relationship (table 23) in the liver in female R-344 rat.
Table 23: retinol and the alpha-tocopherol concentration in rat liver
Lycopene liver in the food
The plain phenol (μ g/gm) of group # mM/kg food mg/kg food retinol (μ g/gm) α-Sheng
Female rats
1??????2.5???????????1280???????1302?????????????47
2??????1.0???????????512????????768??????????????38
3??????0.5???????????256????????858??????????????39
4??????0.25??????????128????????888??????????????46
5??????0.10??????????51?????????555??????????????32
6??????0?????????????0??????????120??????????????13
Male rat
7??????2.5???????????1280???????388??????????????28
8??????1.0???????????512????????119??????????????8
9??????0.5???????????256????????584??????????????23
10?????0.25??????????128????????-????????????????-
11?????0.10??????????51?????????454??????????????14
Matched group 00 16 0
For retinol, between matched group and the high hycopene dosage group 10 times difference is arranged; For alpha-tocopherol, between matched group and the high hycopene dosage group 4 times difference is arranged.The data of male rat also are to be similar to this.Because fill-in contains seldom or do not contain retinol and vitamin E, so all retinols of finding in liver and vitamin must be from AIN-76A food.Therefore, the carotenoid fill-in has changed picked-up and the storage of liver to vitamin A and vitamin E.
Sum up
(1) key component in the lycopene suspension that is made by Fructus Lycopersici esculenti is lycopene (70%), also contains some and comprises α-and other carotenoid of beta-carotene, sigma carotene, lutein, phytoene and phytofluene.
(2) in male or female rats, lycopene does not have adverse effect at used dosage range to food intake or weight increase.
(3) under light at room temperature shone, lycopene can stably be preserved 7 days in food; Under 4 ℃, dark condition, can stably preserve 21 days.
(4) lycopene of the estimator of eating every day nearly 55% is secreted in the feces.
(5) lycopene is easy to absorb in rat liver and store.Also metabolism in liver of lycopene, and metabolic pathway is similar to the approach of finding in the people.Lycopene levels is not linear with food intake in the serum, and this shows that the serum lycopene levels has quite strict homoiostasis control.Lycopene is low 2 orders of magnitude of concentration ratio concentration in liver in serum.In lung, prostate, colon and breast tissue, lycopene is to be detected with ng quantity.Absorb normally relevantly with dosage, least concentration generally is in feeding male and female group of minimum lycopene, especially finds in the liver lycopene concentration.
(6) except that from the liver result who records with high lycopene levels rat feeding, in male and female rats, the glutathione concentrations in whole blood, liver and the kidney is all similar to matched group.
(7) behind the picked-up tomato carotenoids fill-in, the change relevant with dosage taken place with vitamin A concentration in vitamin E in the liver.Do not contain vitamin A because neither contain vitamin E in the fill-in, so the selectivity picked-up from food of vitamin E that the level of finding increases and vitamin A in liver yet.
At last, in the F-344 rat, the whole carotenoid that clearly is incorporated in the lycopene suspension in the AIN-76A half purification food is easy to absorb, enter in the circulation, and is deposited in all test organizations.Lycopene major part in the food is absorbed and is stored in the liver, and lycopene is found the mode metabolism to be similar in the people in liver.Evidence does not show that lycopene has toxicity in used dosage range then.
Though describe and illustrated the present invention by above-mentioned concrete material, operation and embodiment, should be appreciated that the present invention is not limited to the combination and the selected specific operation of enforcement the present invention of these specific materials, material.Numerous variations of these detailed descriptions can imply, and can be understood by those skilled in the art.

Claims (16)

1. carotenoid compositions wherein contains and suspends or be dispersed in lycopene in the medium chain triglyceride.
2. the compositions of claim 1, medium chain triglyceride shown in it are by medium-chain fatty acid and glycerine esterification and obtain.
3. the compositions of claim 2, wherein said medium-chain fatty acid is C 6-C 12Fatty acid.
4. the compositions of claim 3, wherein said medium-chain fatty acid is C 8-C 10Fatty acid.
5. the compositions of claim 2, that wherein said medium-chain fatty acid is selected from is sad, capric acid, lauric acid and their mixture.
6. the compositions of claim 1 wherein contains and accounts for composition weight up to 10% lycopene.
7. the compositions of claim 6 wherein contains the lycopene that accounts for composition weight 4%-5%.
8. the compositions of claim 6 wherein contains and accounts for composition weight up to 98% medium chain triglyceride.
9. the compositions of claim 8 wherein contains the medium chain triglyceride that accounts for composition weight 20%-80%.
10. the compositions of claim 9 wherein contains the medium chain triglyceride that accounts for composition weight 30%-70%.
11. the compositions of claim 8 wherein also contains oil-soluble inhibitor.
12. the compositions of claim 11, wherein said oil-soluble inhibitor are selected from tocopherol, fourth hydroxyanisol, butylated hydroxytoluene, propyl gallate, ethoxy quinoline and ascorbyl palmitate, natural tocopherol, reach their mixture.
13. the compositions of claim 12, wherein said tocopherol be selected from β-, γ-and Delta-Tocopherol and their mixture.
14. the compositions of claim 11, wherein said antioxidant account for composition weight up to 15%.
15. the compositions of claim 14, wherein said antioxidant account for composition weight up to 5%-10%.
16. the method that provides biology can utilize antioxidant comprises effective dose suspended or being dispersed in lycopene in the medium chain triglyceride to host's oral administration.
CN98807515A 1997-05-22 1998-05-20 Carotenoid formulations Pending CN1272059A (en)

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CN102793637A (en) * 2011-05-25 2012-11-28 财团法人纺织产业综合研究所 Skin care product with prompt function and use method thereof
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ATE407693T1 (en) 2004-03-22 2008-09-15 Solvay Pharm Gmbh ORAL PHARMACEUTICAL COMPOSITIONS OF PRODUCTS CONTAINING LIPASE, IN PARTICULAR PANCREATIN, WITH SURFACTANTS
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CN100408030C (en) * 2001-11-12 2008-08-06 利库德天然产品工业有限公司 Method and pharmaceutical preparations for reducing the activity of cells
CN102341003A (en) * 2009-03-05 2012-02-01 巴斯夫欧洲公司 Formulation of astaxanthin derivatives and use thereof II
CN102793637A (en) * 2011-05-25 2012-11-28 财团法人纺织产业综合研究所 Skin care product with prompt function and use method thereof
CN105682684A (en) * 2013-12-11 2016-06-15 健永生技股份有限公司 Vehicular open/close member control device and control method
CN112569212A (en) * 2019-09-30 2021-03-30 富士胶片株式会社 Oily composition, process for producing the same, and soft capsule

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AUPO693397A0 (en) 1997-06-12
JP2001527573A (en) 2001-12-25
ZA984230B (en) 1999-04-20
IL132989A0 (en) 2001-03-19
EP1005339A4 (en) 2004-06-09
EP1005339A1 (en) 2000-06-07
WO1998052561A1 (en) 1998-11-26

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