CN104292091A - Extraction method of capsorubin - Google Patents
Extraction method of capsorubin Download PDFInfo
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- CN104292091A CN104292091A CN201410472563.0A CN201410472563A CN104292091A CN 104292091 A CN104292091 A CN 104292091A CN 201410472563 A CN201410472563 A CN 201410472563A CN 104292091 A CN104292091 A CN 104292091A
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- 235000012658 paprika extract Nutrition 0.000 title claims abstract description 56
- 239000001688 paprika extract Substances 0.000 title claims abstract description 56
- GVOIABOMXKDDGU-LOFNIBRQSA-N (3S,3'S,5R,5'R)-3,3'-dihydroxy-kappa,kappa-carotene-6,6'-dione Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C(=O)C1(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC(=O)C2(C)CC(O)CC2(C)C GVOIABOMXKDDGU-LOFNIBRQSA-N 0.000 title claims abstract description 55
- GVOIABOMXKDDGU-SUKXYCKUSA-N Capsorubin Natural products O=C(/C=C/C(=C\C=C\C(=C/C=C/C=C(\C=C\C=C(/C=C/C(=O)[C@@]1(C)C(C)(C)C[C@H](O)C1)\C)/C)\C)/C)[C@@]1(C)C(C)(C)C[C@H](O)C1 GVOIABOMXKDDGU-SUKXYCKUSA-N 0.000 title claims abstract description 55
- 235000009132 capsorubin Nutrition 0.000 title claims abstract description 55
- GVOIABOMXKDDGU-XRODXAHISA-N (3S,3'S,5R,5'R)-3,3'-dihydroxy-kappa,kappa-carotene-6,6'-dione Chemical compound O=C([C@@]1(C)C(C[C@H](O)C1)(C)C)/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC(=O)[C@]1(C)C[C@@H](O)CC1(C)C GVOIABOMXKDDGU-XRODXAHISA-N 0.000 title claims abstract description 47
- 238000000605 extraction Methods 0.000 title claims abstract description 14
- 239000012071 phase Substances 0.000 claims abstract description 67
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims abstract description 60
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 45
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 27
- 238000010828 elution Methods 0.000 claims abstract description 25
- 238000010829 isocratic elution Methods 0.000 claims abstract description 16
- 239000007791 liquid phase Substances 0.000 claims abstract description 15
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical group CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000000284 extract Substances 0.000 claims abstract description 9
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 238000002390 rotary evaporation Methods 0.000 claims description 26
- 238000004811 liquid chromatography Methods 0.000 claims description 21
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 20
- 229960004756 ethanol Drugs 0.000 claims description 17
- 238000004780 2D liquid chromatography Methods 0.000 claims description 16
- 238000000926 separation method Methods 0.000 claims description 11
- 240000004160 Capsicum annuum Species 0.000 claims description 10
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 claims description 10
- 235000007862 Capsicum baccatum Nutrition 0.000 claims description 10
- 239000001728 capsicum frutescens Substances 0.000 claims description 10
- 239000000377 silicon dioxide Substances 0.000 claims description 10
- 239000002893 slag Substances 0.000 claims description 8
- 150000001408 amides Chemical class 0.000 claims description 6
- 150000002009 diols Chemical class 0.000 claims description 6
- 238000000862 absorption spectrum Methods 0.000 claims description 4
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 4
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000002994 raw material Substances 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 238000004440 column chromatography Methods 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 abstract description 11
- 238000001514 detection method Methods 0.000 abstract description 2
- 238000013375 chromatographic separation Methods 0.000 abstract 2
- JMMWKPVZQRWMSS-UHFFFAOYSA-N isopropanol acetate Natural products CC(C)OC(C)=O JMMWKPVZQRWMSS-UHFFFAOYSA-N 0.000 abstract 2
- 238000002360 preparation method Methods 0.000 description 28
- 239000012141 concentrate Substances 0.000 description 19
- 239000000706 filtrate Substances 0.000 description 18
- 239000006096 absorbing agent Substances 0.000 description 12
- 239000012043 crude product Substances 0.000 description 12
- 239000000047 product Substances 0.000 description 9
- 235000002566 Capsicum Nutrition 0.000 description 8
- 241000208293 Capsicum Species 0.000 description 8
- -1 Capsorubin compound Chemical class 0.000 description 8
- 239000001390 capsicum minimum Substances 0.000 description 8
- 238000004090 dissolution Methods 0.000 description 7
- 238000004587 chromatography analysis Methods 0.000 description 6
- 238000005374 membrane filtration Methods 0.000 description 6
- 238000000967 suction filtration Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 5
- 241001251200 Agelas Species 0.000 description 4
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 235000021466 carotenoid Nutrition 0.000 description 2
- 150000001747 carotenoids Chemical class 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229930014626 natural product Natural products 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- VYIRVAXUEZSDNC-TXDLOWMYSA-N (3R,3'S,5'R)-3,3'-dihydroxy-beta-kappa-caroten-6'-one Chemical compound C([C@H](O)CC=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC(=O)[C@]1(C)C[C@@H](O)CC1(C)C VYIRVAXUEZSDNC-TXDLOWMYSA-N 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- VYIRVAXUEZSDNC-LOFNIBRQSA-N Capsanthyn Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC(=O)C2(C)CC(O)CC2(C)C VYIRVAXUEZSDNC-LOFNIBRQSA-N 0.000 description 1
- PLVBBQBJTBWTDY-XGNSBGGRSA-N Capsochrome Chemical compound O1C2(C)CC(O)CC(C)(C)C2=CC1C(\C)=C/C=C/C=C(\C)/C=C\C=C(/C)\C=C/C=C(/C)\C=C\C(=O)C1(C)CC(O)CC1(C)C PLVBBQBJTBWTDY-XGNSBGGRSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- WRANYHFEXGNSND-LOFNIBRQSA-N capsanthin Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CC(O)CC1(C)C)C=CC=C(/C)C=CC(=O)C2(C)CCC(O)C2(C)C WRANYHFEXGNSND-LOFNIBRQSA-N 0.000 description 1
- 235000018889 capsanthin Nutrition 0.000 description 1
- PLVBBQBJTBWTDY-XMPHPJJSSA-N capsochrome Natural products CC(=C/C=C/C=C(C)/C1OC2(C)CC(O)CC(C)(C)C2=C1)C=CC=C(/C)C=CC=C(/C)C=CC(=O)C3(C)CC(O)CC3(C)C PLVBBQBJTBWTDY-XMPHPJJSSA-N 0.000 description 1
- 229950005499 carbon tetrachloride Drugs 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000021268 hot food Nutrition 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 238000004237 preparative chromatography Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000004223 radioprotective effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 235000021259 spicy food Nutrition 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003815 supercritical carbon dioxide extraction Methods 0.000 description 1
- 238000000194 supercritical-fluid extraction Methods 0.000 description 1
- VZGDMQKNWNREIO-UHFFFAOYSA-N tetrachloromethane Chemical compound ClC(Cl)(Cl)Cl VZGDMQKNWNREIO-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
- C07C45/78—Separation; Purification; Stabilisation; Use of additives
-
- 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
- C07C2601/06—Systems containing only non-condensed rings with a five-membered ring
- C07C2601/08—Systems containing only non-condensed rings with a five-membered ring the ring being saturated
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicines Containing Plant Substances (AREA)
- Cosmetics (AREA)
Abstract
The invention provides an extraction method of capsorubin. The extraction method comprises the following steps: primarily extracting pimiento residues by using anhydrous ethanol and dissolving the primary extract to 30-200mg/mL by using normal hexane; carrying out one-dimensional liquid phase chromatographic separation, namely carrying out isocratic elution as a moving phase A is 0-100% of mixed moving phase in any volume fractions or carrying out gradient elution when the volume fraction of the moving phase A is reduced from 100% to 0%, wherein the phase A is normal hexane and the phase B is isopropanol or ethyl acetate or ethanol; and carrying out two-dimensional liquid phase chromatographic separation on the obtained one-dimensional liquid phase component, namely carrying out isocratic elution when the volume concentration of the moving phase A is 50-100% or carrying out gradient elution when the volume concentration of the moving phase A is reduced from 100% to 50%, so as to obtain the target product capsorubin, wherein the phase A is normal hexane and the phase B is isopropanol or ethyl acetate or ethanol. The method is used for extracting high-purity capsorubin by adopting a two-dimensional liquid chromatographic technology for the first time, and according to detection, the purity of the obtained capsorubin maximally reaches 99%.
Description
Technical field
The present invention relates to food processing technology field, be specifically related to a kind of extracting method of Capsorubin.
Background technology
Capsorubin is a kind of compound in capsicum, belongs to the one of carotenoid.Capsorubin is bright-colored, tinting strength is strong, have and well protect chromatic effect, Capsorubin is as one of haematochrome edible in GB, be widely used in jelly and soy sauce, to the effect of human non-toxic's evil, the compound of the carotenoid in human body can also be increased, there is certain nutritive value, have broad prospects in food applications.
Capsorubin as a kind of antioxidant, to preventing and treat arteriosclerosis and malignant development has certain effect.Meanwhile, the colour developing in various liquid drug, all be unable to do without Capsorubin, and Capsorubin is as a kind of spice, and also have certain effect to pre-radioprotective, research shows, the probability of some national suffers from cancer of liking hot and spicy food is significantly less than other countries.
At present, the extraction about Capsorubin mainly contains solvent extration, oily molten method is extracted and the method for supercritical extraction.Also remain the haematochrome of a great deal of in residue after solvent-extraction process extracts, the product foreign matter content of gained is high, and refine costly, the utilizability of residue is poor, brings difficulty to production; When the molten method of oil extracts capsanthin, oil is more difficult with pigment separated, makes capsicum extraction yield low, is difficult to obtain the high product of look valency; Supercritical carbon dioxide extraction method, this technical matters is simple, energy consumption is low, and extraction solvent easily reclaims, but the product obtained also only just a kind of there is complicated chemical composition extract or thick component (CN 1587321 A, CN 101693785 A, CN 103450702 A).
The Capsorubin purity of producing according to these technology is low, and the quality of product is uncontrollable, so can only be used for food and feed additive industry, cannot be used for high-grade medicine and makeup.
Summary of the invention
Technical problem to be solved by this invention is for above the deficiencies in the prior art, a kind of method extracting Capsorubin from red pepper slag charge is provided, the method adopts two-dimensional liquid chromatography technology to extract high purity Capsorubin first, and its purity of the Capsorubin obtained after testing reaches as high as 99%.
The technical solution adopted in the present invention is:
An extracting method for Capsorubin, the method comprises the following steps:
(1) with red pepper slag charge for raw material, take dehydrated alcohol as Extraction solvent, carry out just carrying 1-5 time, obtain primary extract, it is 30-200mg/mL that primary extract normal hexane carries out being dissolved to concentration;
(2) carry out the separation of one dimension liquid chromatography: the moving phase of employing is two end number mixing moving phase, and wherein A phase is normal hexane, B phase is any one in Virahol, ethyl acetate, ethanol; Type of elution adopts: be that mixed flow equality wash-out or the mobile phase A volume fraction of the arbitrary volume mark of 0-100% is reduced to 0% gradient elution by 100% according to mobile phase A; Sample size is 0.5-5mL/ pin, and flow rate of mobile phase is 10-100mL/min; Detector is UV-detector, determined wavelength 200-400nm, and collect object component according to ultra-violet absorption spectrum, rotary evaporated to dryness, obtains one dimension liquid phase component;
(3) dissolve one dimension liquid phase component by the moving phase of 70-100% normal hexane in step (2), being dissolved to concentration is 10-100mg/mL;
(4) carry out two-dimensional HPLC separation to one dimension liquid phase component: the moving phase of employing is two end number mixing moving phase, and wherein A phase is normal hexane, B phase is any one in Virahol, ethyl acetate, ethanol; Type of elution adopts: mobile phase A volumetric concentration 50-100% isocratic elution or mobile phase A are reduced to 50% gradient elution by 100%; Sample size is 0.5-5mL/ pin, and flow rate of mobile phase is 3-30mL/min; Detector is UV-detector, determined wavelength 200-400nm, and collect object component according to ultra-violet absorption spectrum, rotary evaporation is concentrated into dry, obtains object product Capsorubin.
As preferably, just carry in described step (1) and refer to red pepper slag charge is fully mixed according to solid-liquid ratio/mass volume ratio 1 ︰ (1-20) with dehydrated alcohol, then at the temperature of 30-75 DEG C, extraction 1-3 hour.
As preferably, in described step (2), B phase is ethanol; Type of elution adopts the normal hexane isocratic elution of volume fraction 80-100%.
As preferably, in described step (4), B phase is ethanol; Type of elution adopts the normal hexane isocratic elution of 95-100%.
As preferably, the chromatographic column that in described step (2), one dimension liquid chromatography adopts is positive Silica or Diol or amide chromatogram post, during use, column temperature is room temperature, the chromatographic column that in described step (4), two-dimensional liquid chromatography adopts is for being positive Silica or Diol or amide chromatogram post, and during use, column temperature is room temperature; Further, best chromatogram column combination is that one dimension liquid chromatography adopts positive Silica chromatographic column, and two-dimensional liquid chromatography adopts positive amide chromatogram post simultaneously.
The present invention adopts two-dimensional liquid chromatography technology to carry out extracting Capsorubin from discarded red pepper slag charge.Two-dimensional liquid chromatography technology is separated complex sample by the chromatographic column identical or different by two character, has the separating power and selectivity that improve chromatographic system, shorten preparation time, enriched with trace component, improves the advantages such as preparation sensitivity; Can from the various ingredients of complexity exclusive PCR material, selectively components of interest is prepared; Feasible system automatization, data are reliable, reproducible, are a kind of natural product purification techniques of novelty, more and more applied in the preparation of high-purity natural monomeric compound.Compared with one-dimensional preparative chromatography, two-dimensional liquid chromatography not only has higher peak capacity, and bidimensional has different selectivity.The component simplified through the first dimension can realize complementary separation in the second dimension according to different selectivity, the compound of structural similitude may can arrive in the second dimension and effectively be separated, thus improves efficiency and the compound purity of preparation.
The present invention uses two-dimensional liquid chromatography technology separation and purification Capsorubin from natural product red pepper first, and compared with prior art, the present invention has the following advantages and unusual effect:
(1) chromatographic column is passed through, lysate, flow equal choose reasonable, flow velocity, type of elution, collect the optimization of the processing condition such as section, and first orthogonality tieed up of the peacekeeping second and complementation that reaches is separated, make whole separation purification method practical, operability is high, can be separated from red pepper slag charge and obtain Capsorubin, and the Capsorubin purity obtained is high, 99% is reached as high as through detection, owing to eliminating impurity irrelevant in traditional capsochrome product, the tinting strength of Capsorubin monomer is stronger, anti-oxidant, nourishing function is more obvious, quality product is controlled, may be used for the painted of medicine and the cosmetics of super quality,
(2) from red pepper, extract the method for Capsorubin simple, easy and simple to handle in the present invention, and automation equipment degree is high, just can carry out under normal temperature and pressure, is applicable to the requirement of scale operation;
(3) employing two-dimensional liquid chromatography technology, collection cut batch is stablized, batch-to-batch consistency is high, reproducible, industrial production is convenient to control quality product;
(4) raw material of the present invention is discarded red pepper slag charge, and raw material sources are wide, easily obtain, and both can reach that rubbish utilizes again, the object of protection of the environment, again can the high-grade Industrial products of production high added value, is kill two birds with one stone.
Accompanying drawing explanation
Shown in Fig. 1 is one dimension liquid chromatography separation graph in the embodiment of the present invention 1;
Shown in Fig. 2 is two-dimensional HPLC separation figure in the embodiment of the present invention 1;
Shown in Fig. 3 is that the HPLC of the Capsorubin sample that the embodiment of the present invention 1 obtains tests pure figure.
Embodiment
Below in conjunction with embodiment, the present invention is further described in detail.Iting is noted that following illustrating is all exemplary, being intended to the invention provides further instruction.Except as otherwise noted, all Science and Technology terms that the present invention uses have the identical meanings usually understood with the technical field of the invention personnel.
Embodiment 1:
Take capsicum residue 1.03kg, be dissolved in 4L100% ethanolic soln, 72 DEG C of backflows 1 hour, suction filtration, obtained filtrate and filter residue, filtrate rotary evaporation concentrates, filtrate water after concentrated and extraction into ethyl acetate, concentrate upper solution rotary evaporation, uses 100% n-hexane dissolution after concentrated, concentration is 45mg/mL, carries out the preparation of one dimension liquid chromatography.One dimension liquid chromatography adopts Agela XBP Silica (10 μm
21.2 × 250mm), moving phase adopts normal hexane and ethanol, type of elution: 90% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, sample size is 1mL/ pin, flow rate of mobile phase 16mL/min, as shown in Figure 1, collection Fr.5 peak (retention time 23-27min) is wherein as object component for liquid chromatography separation graph, carry out rotary evaporation to concentrate, for one dimension prepares the crude product of Capsorubin.With 100% n-hexane dissolution Capsorubin crude product, concentration is 20mg/mL, through 0.45 μm of membrane filtration, carry out two-dimensional liquid chromatography preparation, chromatographic column is Acchrom Xamide (10 μm of 20mm × 250mm), moving phase is normal hexane and ethanol, adopt 99% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, sample size is 1mlL/ pin, flow rate of mobile phase 16mL/min, liquid chromatography separation graph as shown in Figure 2, collection peak-2 peak (retention time 4.7-5.5min) is wherein as object component, rotary evaporation, obtain Capsorubin compound, through liquid-phase chromatographic analysis, purity reaches 99%, as shown in Figure 3.
Embodiment 2:
Take capsicum residue 100g, be dissolved in 1L95% ethanol-water solution, 72 DEG C of backflows 1.5 hours, suction filtration, obtained filtrate and filter residue, filtrate rotary evaporation concentrates, filtrate water after concentrated and dichloromethane extraction, concentrate upper solution rotary evaporation, uses 100% n-hexane dissolution after concentrated, concentration is 100mg/mL, carries out the preparation of one dimension liquid chromatography.One dimension liquid chromatography adopts Kromasil Silica (5 μm of 21.2 × 250mm), moving phase adopts normal hexane and ethyl acetate, type of elution: 85-90% normal hexane gradient elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 2mL/ pin, flow rate of mobile phase 18mL/min, collect the cut of 18-19 minute, carry out rotary evaporation and concentrate, for one dimension prepares the crude product of Capsorubin.With 70% normal hexane and 30% dissolve with ethanol Capsorubin crude product, concentration is 30mg/mL, through 0.45 μm of membrane filtration, carry out two-dimensional liquid chromatography preparation, chromatographic column is Acchrom Xamide (10 μm of 20mm × 250mm), moving phase is normal hexane and Virahol, adopt 95-99% normal hexane gradient elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, sample size is 3mlL/ pin, flow rate of mobile phase 12mL/min, collect 5.4-6.0 minute cut, rotary evaporation concentrates, obtain Capsorubin compound, through liquid-phase chromatographic analysis, purity reaches 30%.
Embodiment 3:
Take capsicum residue 500g, be dissolved in 8L70% ethanol-water solution, 72 DEG C of backflows 2 hours, suction filtration, obtained filtrate and filter residue, filtrate rotary evaporation concentrates, filtrate water after concentrated and petroleum ether extraction, concentrate upper solution rotary evaporation, uses 100% n-hexane dissolution after concentrated, concentration is 200mg/mL, carries out the preparation of one dimension liquid chromatography.One dimension liquid chromatography adopts Agela Innoval Silica (10 μm
21.2 × 250mm), moving phase adopts normal hexane and Virahol, type of elution: 95% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 2mL/ pin, flow rate of mobile phase 14mL/min, collect the cut of 29-31 minute, carry out rotary evaporation and concentrate, for one dimension prepares the crude product of Capsorubin.With 80% normal hexane and 20% dissolve with ethanol Capsorubin crude product, concentration is 35mg/mL, through 0.45 μm of membrane filtration, carries out two-dimensional liquid chromatography preparation, and chromatographic column is Agela NH2 (10 μm
21.2 × 250mm), moving phase is normal hexane and ethyl acetate, adopts 97% normal hexane isocratic elution, adopts UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, sample size is 3mL/ pin, flow rate of mobile phase 14mL/min, collects 4.5-5 minute cut, rotary evaporation, obtain Capsorubin compound, through liquid-phase chromatographic analysis, purity reaches 45%.
Embodiment 4:
Take capsicum residue 200g, be dissolved in 2L75% ethanol-water solution, 72 DEG C of backflows 2.5 hours, suction filtration, obtained filtrate and filter residue, filtrate rotary evaporation concentrates, filtrate water after concentrated and extracted with diethyl ether, concentrate upper solution rotary evaporation, uses 100% n-hexane dissolution after concentrated, concentration is 145mg/mL, carries out the preparation of one dimension liquid chromatography.One dimension liquid chromatography adopts YMC Pack SIL (10 μm of 21.2 × 250mm), moving phase adopts normal hexane and ethyl acetate, type of elution: 95%-100% normal hexane gradient elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 2.5mL/ pin, flow rate of mobile phase 18mL/min, collect the cut of 32-33 minute, carry out rotary evaporation and concentrate, for one dimension prepares the crude product of Capsorubin.With 90% normal hexane and 10% dissolve with ethanol Capsorubin crude product, concentration is 15mg/mL, through 0.45 μm of membrane filtration, carries out two-dimensional liquid chromatography preparation, and chromatographic column is Agela Innoval Silica (10 μm
21.2 × 250mm), moving phase is normal hexane and Virahol, adopts 95% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 1mlL/ pin, flow rate of mobile phase 13mL/min, collect 4.5-4.7 minute cut, rotary evaporation, obtains Capsorubin compound, through liquid-phase chromatographic analysis, purity reaches 25%, namely obtains Capsorubin compound.
Embodiment 5:
Take capsicum residue 350g, be dissolved in 6L85% ethanol-water solution, 72 DEG C of backflows 3 hours, suction filtration, obtained filtrate and filter residue, filtrate rotary evaporation concentrates, filtrate water after concentrated and tetrachloromethane extraction, concentrate upper solution rotary evaporation, use 100% n-hexane dissolution after concentrated, concentration is 200mg/mL, carries out the preparation of one dimension liquid chromatography.One dimension liquid chromatography adopts YMC-Pack NH2 (10 μm of 21.2 × 250mm), moving phase adopts normal hexane and Virahol, type of elution: 70%-90% normal hexane gradient elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 1.5mL/ pin, flow rate of mobile phase 13mL/min, collect the cut of 15-17 minute, carry out rotary evaporation and concentrate, for one dimension prepares the crude product of Capsorubin.With 95% normal hexane and 5% dissolve with ethanol Capsorubin crude product, concentration is 40mg/mL, through 0.45 μm of membrane filtration, carry out two-dimensional liquid chromatography preparation, chromatographic column is Kromasil DIol (10 μm of 21.2 × 250mm), moving phase is normal hexane and ethanol, adopt 100% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, sample size is 2mlL/ pin, flow rate of mobile phase 16mL/min, collect 7.5-8 minute cut, rotary evaporation, obtain Capsorubin compound, through liquid-phase chromatographic analysis, purity reaches 70%, namely Capsorubin compound is obtained.
Embodiment 6:
Take capsicum residue 50g, be dissolved in 2L80% ethanol-water solution, 72 DEG C of backflows 1 hour, suction filtration, obtained filtrate and filter residue, filtrate rotary evaporation concentrates, filtrate water after concentrated and n-hexane extraction, concentrate upper solution rotary evaporation, uses 100% n-hexane dissolution after concentrated, concentration is 30mg/mL, carries out the preparation of one dimension liquid chromatography.One dimension liquid chromatography adopts YMC-Pack NH2 (10 μm of 21.2 × 250mm), moving phase adopts normal hexane and ethanol, type of elution: 80% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, and sample size is 3mL/ pin, flow rate of mobile phase 16mL/min, collect the cut of 13-14 minute, carry out rotary evaporation and concentrate, for one dimension prepares the crude product of Capsorubin.With 85% normal hexane and 15% dissolve with ethanol Capsorubin crude product, concentration is 22mg/mL, through 0.45 μm of membrane filtration, carry out two-dimensional liquid chromatography preparation, chromatographic column is Kromasil DIol (10 μm of 21.2 × 250mm), moving phase is normal hexane and Virahol, adopt 95%-100% normal hexane isocratic elution, adopt UV6000 UV-detector 400nm Selective absorber wavelength, preparation temperature is room temperature, sample size is 3mL/ pin, flow rate of mobile phase 12mL/min, collect 6.3-6.7 minute cut, rotary evaporation, obtain Capsorubin compound, through liquid-phase chromatographic analysis, purity reaches 40%.
The material that the embodiment of the present invention relates to, reagent and experimental installation, if no special instructions, be the common commercially available prod meeting food-processing.
The above, be only the preferred embodiments of the present invention, should be understood that; for those skilled in the art; under the prerequisite not departing from core technology of the present invention, can also make improvements and modifications, these improvements and modifications also should belong to scope of patent protection of the present invention.Any change in the implication suitable with claims of the present invention and scope, all should think to be included in the scope of claims.
Claims (7)
1. an extracting method for Capsorubin, is characterized in that comprising the following steps:
(1) with red pepper slag charge for raw material, take dehydrated alcohol as Extraction solvent, carry out just carrying 1-5 time, obtain primary extract, it is 30-200mg/mL that primary extract normal hexane carries out being dissolved to concentration;
(2) carry out the separation of one dimension liquid chromatography: the moving phase of employing is two end number mixing moving phase, and wherein A phase is normal hexane, B phase is any one in Virahol, ethyl acetate, ethanol three kinds; Type of elution adopts: be that mixed flow equality wash-out or the mobile phase A volume fraction of the arbitrary volume mark of 0-100% is reduced to 0% gradient elution by 100% according to mobile phase A; Sample size is 0.5-5mL/ pin, and flow rate of mobile phase is 10-100mL/min; Detector is UV-detector, determined wavelength 200-400nm, and collect object component according to ultra-violet absorption spectrum, rotary evaporated to dryness, obtains one dimension liquid phase component;
(3) dissolve one dimension liquid phase component by the moving phase of 70-100% normal hexane in step (2), being dissolved to concentration is 10-100mg/mL;
(4) carry out two-dimensional HPLC separation to one dimension liquid phase component: the moving phase of employing is two end number mixing moving phase, and wherein A phase is normal hexane, B phase is any one in Virahol, ethyl acetate, ethanol three kinds; Type of elution adopts: mobile phase A volumetric concentration 50-100% isocratic elution or mobile phase A are reduced to 50% gradient elution by 100%; Sample size is 0.5-5mL/ pin, and flow rate of mobile phase is 3-30mL/min; Detector is UV-detector, determined wavelength 200-400nm, and collect object component according to ultra-violet absorption spectrum, rotary evaporation is concentrated into dry, obtains object product Capsorubin.
2. the method for the extracting method of a kind of Capsorubin according to claim 1, it is characterized in that: just carry in described step (1) and refer to red pepper slag charge is fully mixed according to solid-liquid ratio 1 ︰ (1-20) with dehydrated alcohol, then, at the temperature of 30-75 DEG C, 1-3 hour is extracted.
3. the method for the extracting method of a kind of Capsorubin according to claim 1, is characterized in that: in described step (2), B phase is ethanol; Type of elution adopts the normal hexane isocratic elution of volume fraction 80-100%.
4. the method for the extracting method of a kind of Capsorubin according to claim 1, is characterized in that: in described step (4), B phase is ethanol; Type of elution adopts the normal hexane isocratic elution of 95-100%.
5. the method for the extracting method of a kind of Capsorubin according to claim 1, is characterized in that: the chromatographic column that in described step (2), one dimension liquid chromatography adopts is positive Silica or Diol or amide chromatogram post, and during use, column temperature is room temperature.
6. the method for the extracting method of a kind of Capsorubin according to claim 1, is characterized in that: the chromatographic column that in described step (4), two-dimensional liquid chromatography adopts is for being positive Silica or Diol or amide chromatogram post, and during use, column temperature is room temperature.
7. the method for the extracting method of a kind of Capsorubin according to claim 5 or 6, is characterized in that: described one dimension liquid chromatography adopts positive Silica chromatographic column, and two-dimensional liquid chromatography adopts positive amide chromatogram post simultaneously.
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| CN105693583A (en) * | 2015-12-30 | 2016-06-22 | 晨光生物科技集团股份有限公司 | Method for removing harmful ingredients in carotenoid |
| CN105693583B (en) * | 2015-12-30 | 2018-02-09 | 晨光生物科技集团股份有限公司 | The method of composition is endangered in a kind of removal carotenoid |
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