CN101845236A - Method for extracting melanin from black waxy corn - Google Patents
Method for extracting melanin from black waxy corn Download PDFInfo
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Abstract
The invention belongs to a method for extracting melanin, in particular to a method for extracting melanin from black waxy corn, which is characterized in that black waxy corn on the cob is adopted as a raw material, supercritical carbon dioxide is used as an extracting agent, and less entrainer is used. Compared with the prior art, the extraction rate can reach 14.32 percent. Test results show that the preferred process is that: extracting pressure is 30MPa, temperature is 50 DEG C, time is 90 minutes, flow rate of carbon dioxide is 120 L/h, the entrainer is of absolute ethyl alcohol, and the granularity of the raw material is 110 meshes. After ethanol in the obtained extracting liquid is volatilize, water is added to dissolve the obtained extracting liquid, after being purified by macroporous absorption resin and being washed by distilled water to be colorless, ethanol solution with the concentration of 50 to 70 percent is used as eluant to elute, the obtained ethanol eluant liquid is decompressed and concentrated to obtain extract, and the extract is dried to obtain the melanin finished product. The purity of the melanin is remarkably improved compared with that before the purification.
Description
[technical field]
The invention belongs to melanic extracting method, more particularly, relate to a kind of melanic method of from black waxy corn, extracting.
[background technology]
Black waxy corn belongs to Gramineae Zea (Gramineae ZeaL.), originates in the mountain area, South America, and its size is identical with conventional corn, but all near black such as its seed and bract, in China, black waxy corn is a kind of cereal of a large amount of plantations in each department, it is as black rice, and Semen Sesami Nigrum etc. are equally originated very abundant.Effect anti-oxidant, that remove free radical that the melanochrome that extracts from the whole dried black waxy corn of fringe has in human body has functions such as the body immunity of increasing, anti-ageing, vessel softening, cancer-resisting.This pigment and nitrous acid ratio can suppress 99% the carcinogenic formation of nitrous acid at 1: 3 o'clock.
The domestic and international market increases day by day to the natural pigment requirement at present, is the rate of growth of 4%-5% every year.Although synthetic colour lovely luster, strong coloring force, stable in properties, synthetic colour does not have nutrition, has chemical toxicity.The melanochrome that extracts from the whole dried black waxy corn of fringe is the pure natural flavonoid compound, belongs to anthocyanin class pigment, for water-soluble.Have good bio-physiological activity, good market outlook are arranged in the health care of food industry.Report about melanic extraction in the black Semen Maydis is existing at present, but do not see and use CO
2Melanochrome and detailed process for refining report in the whole dried black waxy corn of fringe of supercritical extraction.
[summary of the invention]
The object of the present invention is to provide a kind of melanic method of from black waxy corn, extracting, particularly use supercritical liquid extraction technique from the whole dried black waxy corn of fringe, to extract melanic method.
The present invention solves the problems of the technologies described above by the following technical programs:
A kind ofly from black waxy corn, extract melanic method, it is characterized in that may further comprise the steps:
A. raw materials pretreatment: will put in order the dried black waxy corn of fringe through sorting, drying, being ground into fineness with high speed disintegrator is 100-120 purpose powder, and the dehydrated alcohol that adding and described black waxy corn powder etc. are heavy is pressed 1: 1 mixed;
B. use supercritical CO
2Extraction melanochrome:
The mixture that step a is obtained drops in the extraction kettle, when described extraction temperature in the kettle reaches 40-60 ℃, opens CO
2Gas cylinder is supplied gas to described extraction kettle, when described extraction kettle internal pressure reaches 10-35Mpa, regulates CO
2Flow reaches 120L/h, is extracting 90-150min under the constant temperature and pressure under said temperature and the pressure, and certain hour is collected extract from the extraction-container exit at interval;
C. the pre-treatment of resin: get new D101, D201 or AB-8 macroporous adsorbent resin, with 95% alcohol-pickled a few hours, clean with a large amount of distilled water, under agitation described resin is filled in the chromatography column with wet method then, water is emitted after forming resin layer etc. described resin settled, the ethanol with 95% is by described resin layer, until effusive liquid till no white opacity phenomenon behind the adding distilled water, with same flow velocity described resin layer is washed till neutrality with distilled water again, standby;
D. melanochrome purifying: with step b gained extract solution, vapor away ethanol wherein, be dissolved in water, this aqueous solution is pumped into described chromatography column, earlier with distilled water be eluted to colourless after, ethanolic soln with 50%～70% concentration carries out wash-out as eluent, and resulting ethanol eluate gets medicinal extract through concentrating under reduced pressure, obtains the melanochrome finished product after the drying.
Principle of the present invention is: supercritical liquid extraction technique is the new technology that development in recent years is got up, and is being used more and more widely aspect the active skull cap components extraction, and this technology is with the extraction feed extraction kettle of packing into.The employing carbonic acid gas is a supercritical solvent.Carbon dioxide is condensed into liquid through heat exchanger, with force (forcing) pump boost in pressure to the required pressure of technological process (emergent pressure that should be higher than carbonic acid gas), simultaneously attemperation makes it become supercritical carbon dioxide fluid.CO 2 fluid enters from the extraction kettle bottom as solvent, fully contacts with being extracted material, and selective dissolution goes out required chemical ingredients.The high-pressure carbon dioxide fluid that contains the dissolution extraction thing is depressured to be lower than through throttling valve and enters separating still (claiming extraction-container again) below the carbonic acid gas emergent pressure, owing to sharply descending, carbonic acid gas solubleness separates out solute, automatically be separated into solute and carbon dioxide two parts, the former is the process product, regularly emit from the separating still bottom, the latter is a circulate carbon dioxide gas, is condensed into carbon dioxide liquid recirculation through over-heat-exchanger and uses.Whole sepn process is to utilize CO 2 fluid organism to be had the solubleness of special increase under supercritical state, and under the subcritical state to the undissolved substantially characteristic of organism, CO 2 fluid is constantly circulated between extraction kettle and separating still, thereby will need the component of separation and Extraction from raw material, to separate effectively.The present invention is with supercritical CO
2As extraction agent, be aided with a small amount of entrainment agent, compared with prior art, percentage extraction can reach 14.32%, has increased substantially melanic yield.
The invention has the advantages that: the present invention has extraction yield height, good product purity, simple, the low power consumption and other advantages of flow process.
[embodiment]
1. the starting material of black Semen Maydis are handled and are used supercritical CO
2Extract melanic whole process:
1.1.1 raw materials pretreatment
The dried black waxy corn of whole fringe utilizes high speed disintegrator to be ground into different fineness through sorting, drying, presses 1: 1 mixed of the dried black waxy corn fine powder of dehydrated alcohol and whole fringe.
1.1.2 supercritical CO
2The whole fringe of extraction is done the method for Pigment in Black Glutinous Maize
Accurately take by weighing the above-mentioned sample handled well of 500g and drop in the extraction kettle, but temperature is opened CO when reaching preset temperature respectively
2Gas cylinder is supplied gas, and when pressure reaches predetermined pressure, regulates flow, begins extraction and calculating extraction time under the constant temperature and pressure, and certain hour is collected extract from the extraction-container exit at interval.
1.2 result and analysis
1.2.1 the selection of extracting pressure
Experimental result (data see Table 1) shows that at 10-35Mpa, whole fringe is done Pigment in Black Glutinous Maize to be increased with the pressure rising, and this is because supercritical CO
2Fluid has very big compressibility, and under the certain temperature, pressure increases, and the density of supercutical fluid increases, and the solubleness of solute in fluid increases, and pressure increase has simultaneously also reduced intermolecular mass transfer distance, has increased the mass-transfer efficiency between solvent and solute.But when reaching 30Mpa, it is maximum that the pigment yield almost reaches, and this moment, pressure increased again, and the whole dried black waxy corn medicinal extract of fringe rate can increase, but because the content of pigment is limited, increasing part is impurity component, and therefore best extracting pressure is 30Mpa.
Table 1 extracting pressure is done the influence of Pigment in Black Glutinous Maize extraction yield to whole fringe
1.2.2 the selection of extraction temperature
Under the constant voltage, temperature raises, supercutical fluid reduces than degree, cause the fluid solvent effect to descend, make solute descend, be unfavorable for extraction in its solubleness, but temperature is the function of solute volatility, improve the vapour pressure that temperature can improve solute exponentially, thereby increase the solubleness of solute, improve extraction efficiency.By experimental data (table 2) as can be known, with the raising of extraction temperature, percentage extraction increases, but extraction temperature is when being higher than 50 ℃, and percentage extraction descends, and therefore, best extraction temperature is 50 ℃.
Table 2 extraction temperature is done the influence that Pigment in Black Glutinous Maize is extracted to whole fringe
1.2.3 the selection of extraction time
Whole supercritical extraction had both comprised transhipment, had diffusion again, and the static extracting time that keeps certain can be improved extraction efficiency.Therefore we have carried out 5 groups of tests 30,60,90,120, under the 150min, and result's (seeing Table 3) shows: along with the increase of time, effect of extracting is become better and better, and surpasses 90min when the time, and extraction results changes not quite, and therefore, optimum extraction time is 90min.
The table 3 extraction time is done the influence that Pigment in Black Glutinous Maize is extracted to whole fringe
1.2.4CO
2The selection of flow
CO
2Flow is little to the influence of extraction yield, and enlargement discharge has then increased the extraction times of solvent to raw material, can shorten the extraction time, and on the other hand, flow velocity is accelerated, and its consumption increases, and in conjunction with the actual energy saving, we are to CO
2Flow is investigated, and result's (seeing Table 4) shows that optimal flow rate is 120L/h.
Table 4CO
2Whole fringe is done the influence that Pigment in Black Glutinous Maize is extracted
1.2.5 the selection of entrainment agent
The solvency action of carbon dioxide upercritical fluid extraction: supercritical CO
2Fluidic polarity is generally between normal hexane and chloroform, under supercritical state, CO2 is very big to the dissolving power difference of different solutes, polarity, boiling point and the molecular weight of this and solute are closely related, in general following rule is arranged: lipotropy, low boiling point component can extract below 104KPa, as volatile oil, hydrocarbon, ester, lactone, ether, epoxy compounds etc., as the aroma component in natural phant and the fruit, as low boiling ester class in Terpane, thymol, the hops etc.; The polar group of compound (as-OH ,-COOH etc.) the more, then more difficult extraction.Therefore extracting polarity than the great achievement timesharing, the general a spot of polar solvent (entrainment agent) that adds, to change effect of extracting, in the test, we have investigated ethyl acetate, acetone, water, dehydrated alcohol, 95% ethanol and five kinds of solvents of 0.1mol/L hydrochloric acid (1: 1), and result's (seeing Table 5) shows, and are the highest with dehydrated alcohol and 95% ethanol and 0.1mol/L hydrochloric acid (1: 1) extraction efficiency, consider the corrodibility of actually operating hydrochloric acid, we have selected dehydrated alcohol as entrainment agent.
Table 5 entrainment agent is done the influence that Pigment in Black Glutinous Maize is extracted to whole fringe
1.2.6 the selection of entrainment agent consumption
Add a certain amount of entrainment agent and help extraction, but the entrainment agent of crossing a large amount will make the extraction state carry out under a kind of non-critical state, often do not reach the ideal effect of extracting, therefore the amount of entrainment agent will reach requirement, we have tested 4 amounts (1: 0.2,1: 0.5,1: 1,1: 2), result's (seeing Table 6) shows, adds entrainment agent effect of extracting the best of 1: 1.
Table 6 entrainment agent consumption is done the influence that Pigment in Black Glutinous Maize is extracted to whole fringe
1.2.7 the selection of raw material granularity
We have taked granularity is 80,90,100,110,120 to have carried out 5 groups of tests now, and test-results (seeing Table 7) shows that along with the increase of granularity, effect of extracting is become better and better.In general raw material granularity is more little, and solute path to the supercutical fluid transmission from raw material is shorter, and the surface-area that contacts with supercutical fluid is bigger, helps extraction.But granularity also should not be too little, and meticulous granularity had both increased the difficulty of pulverizing, and causes extraction kettle egress filtering net to stop up again easily, produces huge pressure difference and destruction equipment, in therefore testing we to have selected granularity be 110 orders.
Table 7 raw material granularity is done the influence that Pigment in Black Glutinous Maize is extracted to whole fringe
1.3 result and discussion
The present invention discloses the employing supercritical CO first
2The melanic processing condition of extraction from the whole dried black waxy corn of fringe, result of study show that the melanic optimum process condition of extraction is pressure 30Mpa, 50 ℃ of temperature, time 90min, CO
2Flow is that 120L/h, entrainment agent are dehydrated alcohol (solid-to-liquid ratio is 1: 1), fineness 110 orders, and percentage extraction can reach 14.32% under optimum process condition.
2. melanic purifying
Macroporous adsorbent resin originates from the seventies in 20th century, it is a kind of polymer absorbant with porous three dimensional structure synthetic, utilize its huge surface-area to carry out physical adsorption, in actual applications some molecules close with its skeleton structure such as aromatic series ring-type compound are had very strong adsorptive power, be widely used in the extraction separation of macromolecular cpds such as activeconstituents in pharmacy and the natural phant such as saponin(e, flavones, lactone, alkaloid.
The pre-treatment of macroporous resin: get commercially available new D101, D201, AB-8, five kinds of macroporous adsorbent resins such as H103, S-8, used for 95% alcohol-pickled a few hours respectively, clean with a large amount of distilled water.Under agitation resin is joined in the chromatography column with wet method, water is emitted etc. after the resin settled again, the ethanol with 95% is by resin layer, and adding an amount of distilled water to effluent liquid does not have the white opacity phenomenon, is washed till neutrality with distilled water with same flow velocity again, standby.
2.1 the mensuration of purifying rate
Get melanochrome solution and measure absorption peak in the 400-800nm scope, by the result as can be known, melanochrome has maximum absorption at the 526nm place, the reference report
[8]The material that can judge the 526nm place is the anthocyanin class material, and solution behind the purifying is measured absorbancy as the purifying index in the 526nm place.
2.2 result and analysis
2.2.1 the screening of macroporous adsorbent resin
Get the melanochrome extract of supercritical extraction, fling to ethanol, be dissolved in water.With concentration is the melanochrome extract aqueous solution good resin column of pre-treatment by 5 kinds of different models respectively of 50mg/ml, carries out adsorptive capacity and measures.Effluent liquid utilizes melanochrome charateristic avsorption band 526nm place to measure extinction density (A) value, has judged whether that melanochrome flows out, when the positive appears in the result, show that existing melanochrome flows out, resin absorption reaches capacity, and stops to go up sample, the record upper column quantity calculates the melanochrome adsorptive capacity, and unit is mg/ml.The results are shown in Table 8.
Table 8 different resins is to melanic adsorptive capacity in the whole dried black waxy corn of fringe
Resin column behind the last sample washes with water earlier to colourless, use 50% ethanol elution instead to colourless, collect 50% ethanol eluate, through concentrating under reduced pressure, vacuum-drying gets red crystals, measure absorbance (A) value, result's (seeing Table 9) shows that the D101 resin has melanochrome in the black waxy corn selects adsorptivity and desorption ability preferably, so select for use the D101 resin to carry out the technical study of this experiment.
Table 9 different resins is to melanic parsing amount in the whole dried black waxy corn of fringe
2.2.2 the selection of eluent and concentration thereof
Get a certain amount of black waxy corn extract aqueous solution, upper prop, earlier with distilled water be eluted to colourless after, ethanolic soln with 50mL 10%～80% concentration carries out wash-out successively, collect the Different concentrations of alcohol elutriant and be settled to 50ml, get wherein and to measure melanic absorbance (A) value after the 3ml elutriant is diluted to 10ml, to determine the eluant strength of the best.
Table 10 different concentration ethanol is to melanic wash-out influence in the whole dried black waxy corn of fringe
By test-results (table 10) as can be known, alcohol concn increases, and relative eluted product also increases thereupon, but by fixing of melanic amount, concentration surpasses 50%, and impurity also increases thereupon, melanic absorbance (A) diminishes, so select the eluent of the ethanol of 50% concentration as resin.
2.2.3 the comparison before and after the purification with macroreticular resin melanochrome
Get a certain amount of black waxy corn extract aqueous solution and before and after upper prop, measure melanic absorbance (A) value, the results are shown in Table 11.
Comparison in the whole dried black waxy corn of fringe of table 11 before and after the melanochrome purifying
By last table data as can be known, the A value behind the melanochrome upper prop is before greater than upper prop, and the A value is big more, illustrates that pigment purity is high more, can judge that thus the purity after melanochrome is purified obviously improves.
2.3 melanic conclusion of purification with macroreticular resin and analysis
The melanic technology of purification with macroreticular resin is defined as: get the black waxy corn extract aqueous solution and separate through macroporous adsorption resin chromatography, resin and raw material ratio are 1: 1 (W/W), elder generation's water is eluted to colourless, use 50% ethanol as the eluent wash-out again, the concentrating under reduced pressure ethanol eluate gets medicinal extract, the dry yield that gets product is 7.65%, and content is 98.1%.
Although illustrated and described embodiments of the invention, for the ordinary skill in the art, be appreciated that without departing from the principles and spirit of the present invention and can carry out multiple variation, modification, replacement and modification to these embodiment, scope of the present invention is limited by claims and equivalent thereof.
Claims (6)
1. one kind is extracted melanic method from black waxy corn, it is characterized in that may further comprise the steps:
A. raw materials pretreatment: will put in order the dried black waxy corn of fringe through sorting, drying, being ground into fineness with high speed disintegrator is 100-120 purpose powder, and the dehydrated alcohol that adding and described black waxy corn powder etc. are heavy is pressed 1: 1 mixed;
B. use supercritical CO
2Extraction melanochrome:
The mixture that step a is obtained drops in the extraction kettle, when described extraction temperature in the kettle reaches 40-60 ℃, opens CO
2Gas cylinder is supplied gas to described extraction kettle, when described extraction kettle internal pressure reaches 10-35Mpa, regulates CO
2Flow reaches 120L/h, is extracting 90-150min under the constant temperature and pressure under said temperature and the pressure, and certain hour is collected extract from the extraction-container exit at interval;
C. the pre-treatment of resin: get new D101, D201 or AB-8 macroporous adsorbent resin, with 95% alcohol-pickled a few hours, clean with a large amount of distilled water, under agitation described resin is filled in the chromatography column with wet method then, water is emitted after forming resin layer etc. described resin settled, the ethanol with 95% is by described resin layer, until effusive liquid till no white opacity phenomenon behind the adding distilled water, with same flow velocity described resin layer is washed till neutrality with distilled water again, standby;
D. melanochrome purifying: with step b gained extract solution, vapor away ethanol wherein, be dissolved in water, this aqueous solution is pumped into described chromatography column, earlier with distilled water be eluted to colourless after, ethanolic soln with 50%～70% concentration carries out wash-out as eluent, and resulting ethanol eluate gets medicinal extract through concentrating under reduced pressure, obtains the melanochrome finished product after the drying.
2. according to the described a kind of melanic method of from black waxy corn, extracting of claim 1, it is characterized in that in the step a of described method that will putting in order the dried black waxy corn of fringe, to be ground into fineness with high speed disintegrator be 110 purpose powder.
3. according to the described a kind of melanic method of extracting from black waxy corn of claim 1, the extraction temperature that it is characterized in that the step b of described method is 50 ℃.
4. according to the described a kind of melanic method of from black waxy corn, extracting of claim 1, it is characterized in that extracting when extraction kettle internal pressure among the step b of described method reaches 30Mpa.
5. according to the described a kind of melanic method of from black waxy corn, extracting of claim 1, it is characterized in that the extraction time of the step b of described method is 90min.
6. according to the described a kind of melanic method of from black waxy corn, extracting of claim 1, it is characterized in that in the steps d of described method with the ethanol of 50% concentration eluent as resin.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102352121A (en) * | 2011-11-02 | 2012-02-15 | 安徽大诚明农业科技有限公司 | Method for extracting black pigment from black waxy corn complete cobs |
| CN104705570A (en) * | 2015-04-09 | 2015-06-17 | 王振祥 | Compound ultrafine black glutinous rice flour |
| CN107099166A (en) * | 2017-04-14 | 2017-08-29 | 罗莱生活科技股份有限公司 | A kind of method of supercritical carbon dioxide extracting banana skin dyestuff and the purposes of banana skin dyestuff |
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| CN1325924A (en) * | 2001-06-25 | 2001-12-12 | 王力田 | Process for extracting melanin |
| CN1367208A (en) * | 2002-03-04 | 2002-09-04 | 大庆石油管理局 | Process for extracting carotenol by using supercritical CO2 extraction |
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| CN1325924A (en) * | 2001-06-25 | 2001-12-12 | 王力田 | Process for extracting melanin |
| CN1367208A (en) * | 2002-03-04 | 2002-09-04 | 大庆石油管理局 | Process for extracting carotenol by using supercritical CO2 extraction |
| CN1844116A (en) * | 2006-05-10 | 2006-10-11 | 江南大学 | Extraction separation purification and identification of flavonoid monomers in oriental blueberry melanin |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102352121A (en) * | 2011-11-02 | 2012-02-15 | 安徽大诚明农业科技有限公司 | Method for extracting black pigment from black waxy corn complete cobs |
| CN104705570A (en) * | 2015-04-09 | 2015-06-17 | 王振祥 | Compound ultrafine black glutinous rice flour |
| CN107099166A (en) * | 2017-04-14 | 2017-08-29 | 罗莱生活科技股份有限公司 | A kind of method of supercritical carbon dioxide extracting banana skin dyestuff and the purposes of banana skin dyestuff |
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