CN103421611B - Method for extracting effective constituents from holy basil - Google Patents
Method for extracting effective constituents from holy basil Download PDFInfo
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- CN103421611B CN103421611B CN201310408832.2A CN201310408832A CN103421611B CN 103421611 B CN103421611 B CN 103421611B CN 201310408832 A CN201310408832 A CN 201310408832A CN 103421611 B CN103421611 B CN 103421611B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02P20/00—Technologies relating to chemical industry
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Abstract
The invention relates to a method for extracting effective constituents from holy basil. Newly-collected holy basil is used as raw materials, and water-soluble effective constituent extractive and essential oil part extractive are obtained through aging treatment, enzymolysis, resin adsorption, holy basil log smoke curing and essential oil extraction through supercritical fluid. An ancient folk technology and a modern leading-edge process are organically bonded in the method, the extracted effective constituents are free of residual solvents, aroma components are not damaged, the effective constituents are completely extracted, and extraction yield is high.
Description
Technical field
The present invention relates to a kind of extracting method of active ingredient of natural plant, particularly a kind of from Ocimum basilicum L. the method for extracting effective components.
Background technology
Ocimum basilicum L. also claims Lysimachia foenum-graecum (Lysimachia foenum-graecum Hance) for Primulaceae Lysimachia plant, and main product is in Yao Autonomous County of Jinxiu, Guangxi Zhuang Autonomous Region, Yao Autonomous County of Jianghua, Hunan Province.The name of Ocimum basilicum L. begins to be loaded in " the good book on Chinese herbal medicine of helping ", " Mingyi Bielu " smoke grass.Song's " figure is through book on Chinese herbal medicine " is carried: " Ocimum basilicum L., modern lake, ridge Zhu Zhou all have it, give birth to wetland more.Leaf is as fiber crops, relative between two, and stem side's gas is as the seedling of xiongqiong, and often to bloom mid-July, to fragrant, also, or cloud, also this is also to smoke grass for ancient so-called smoked grass ".Ocimum basilicum L. contains more organic acid and aromatic series phenol, alcohol, aldehyde, ketone and derivative thereof.This compounds volatilization is slow, fragrant odour, after news, feel immediately easily, and be the stronger major cause of the lasting fixation power of fragrance in Ocimum basilicum L., have " fragrant king's title ", be the expensive perfume base of a kind of generic name.Its effective constituent is essential oil part and water-soluble portion.Essential oil part is containing the composition of volatile oil, be mainly oxygenatedchemicals class, oxygenatedchemicals accounts for 70% of total composition, its main component is 9,12-18-diolefinic acid, margaric acid, pohytol, butyl phthalate, the undecyl aldehyde etc., be widely used in the industries such as medicine, spices, food, tobacco, weaving, daily-use chemical industry.And in water-soluble portion, contain the triterpenoid saponin compounds of serial oleanane type, and there is immunomodulatory, the physiologically active such as antitumor, antiviral, antimycotic, anticancer, be a compounds that has application prospect.
But, prior art is mainly solvent method and steam distillation to the extracting method of Ocimum basilicum L. effective constituent, solvent method can produce dissolvent residual, and to the essential oil part in extract and water-soluble portion non-selectivity, solvent recuperation process temperature is higher, and destructible aroma component also may have certain pollution to environment, technical process is long, length consuming time.Steam distillation temperature is high, destructible aroma component, and especially to rare spice, bottom note material is difficult to propose, and extraction yield is low, and resource is caused to waste.For this reason, open day on April 30th, 2003, Chinese invention patent application publication number CNl414077A.Disclosing a kind of extracting method of high quality lingxiang grass purified oil, is to have adopted supercritical CO
2fluid is that extraction solvent extracts Lysimachia foenum-graecum raw material, and extracting solution is again through cryogenic freezing dewaxing, and high speed centrifugation separates and obtains high quality lingxiang grass purified oil, best extractant condition is: extracting pressure is 12MPa, temperature is 35 DEG C, and the time is 1.5h, and flow is 180L/h; Separation condition is: pressure is 5MPa, and temperature is 45 DEG C; In frozen centrifugation dewaxing, freezing temp is-15--20 DEG C, and rotating speed is 4000-5000 rev/min.But this technology is by simple supercritical CO
2fluid extraction means, are only confined to extract small molecules amount essential oil part, and the effective constituent of macromolecule water-soluble portion is but left in the basket; In addition, because Ocimum basilicum L. is a kind of Primulaceae Lysimachia plant, organic acid, aromatic series phenol, alcohol, aldehyde, ketone and the derivative thereof etc. in its effective constituent are containing existing between plant in cell and in intercellular pectin substance, CNl414077A CO
2fluid extraction, to raw material processing be will be first by raw material through pulverizing, but the particle of this pulverizing can not very carefully injure cell, meticulous powder not only expends power source, and forms and stick with paste diploma thing and stop up conversely CO
2fluid extraction.So exist extracts active ingredients imperfect, extract yield is lower than 2.0%.
Summary of the invention
The technical problem to be solved in the present invention be to provide a kind of from Ocimum basilicum L. the method for extracting effective components.This extracting method collection technology among the people in ancient times and modern forward position technique organically combine, and gained extracting effective components no solvent residue, do not destroy aroma component, and extracts active ingredients is complete, and extract yield is high.
The present invention solves the problems of the technologies described above by the following technical programs:
A kind of from Ocimum basilicum L. the method for extracting effective components, taking the Ocimum basilicum L. of fresh collection as raw material, through smoke → supercritical fluid extraction of ripening → enzymolysis → resin absorption → Ocimum basilicum L. log essential oil, obtain aqueous soluble active constituent extract and essential oil extracting section thing; Specifically comprise the steps:
(1) ripening
Gather fresh Ocimum basilicum L. herb, wash away sand, after draining, ageing 3-4 hour, to forge native mud pathogenic dryness;
(2) enzymolysis
Forge the herb after native mud pathogenic dryness, being cut into long broken section of 30-50mm puts in stainless steel vessel, till adding deionized water to alfalfa section and being just immersed in water, add 1000-1500U cellulase and 600-800U polygalacturonase by every kilogram of alfalfa section, with hydrochloric acid adjust pH to 4. 8, fully stir, 42 DEG C of water bath with thermostatic control 4 h, let cool to room temperature, extremely neutral with 0.2mol/L sodium hydroxide solution adjust pH, filtering separation;
(3) resin absorption
Step (2) parting liquid resin absorption; Resin after washing absorption until effluent liquid approach colourless, then the alcoholic solution wash-out that is 50-100% by concentration, collection elutriant, drying, obtains the Ocimum basilicum L. aqueous soluble active constituent extract of saponin content >=10%, this extract is specially triterpenoid saponin;
(4) Ocimum basilicum L. log is smoked
The Ocimum basilicum L. slag that step (2) separates, rinses through water, drains; Cut that to get on mountain bamboo several, length 3-5m, the two halves of riving, wash in a pan empty; The Ocimum basilicum L. slag draining is evenly laid in and is washed in a pan in half empty sheet bamboo, stay a little space ventilative, cover other half sheet bamboo, fix as bamboo folder cylinder; Each bamboo folder cylinder is controlled and separated slag loading level is 5kg; The bamboo folder cylinder of the full Ocimum basilicum L. slag of folder is emitted on bamboo bed, under bamboo bed, is paved with log dried firewood or igniting and fire-cures, while fire-cureing, will often stir the position of bamboo folder cylinder, make it uniform drying; The time of fire-cureing will be seen firepower size and determine, generally in 6-8 hour, is advisable; After fire-cureing, can take out the Ocimum basilicum L. slag fire-cureing and be placed on the ground, spray a little moisture, good with straw mulch, moisture is dissipated, after dry;
(5) supercritical fluid extraction essential oil
By the Ocimum basilicum L. slag fire-cureing, adopt supercritical CO
2fluid is that extraction solvent extracts, and extracting solution separates and obtains essential oil extracting section thing through high speed centrifugation again; Supercritical CO
2extractant condition is: extracting pressure is 17-20MPa, and temperature is 40-42 DEG C, and the time is 2-3h, and flow is 250-300L/h; The condition of centrifugation is: rotating speed 6000-7000 rev/min, temperature is normal temperature.
" U " Shi Meihuo unit of described step (2), the 1 U enzyme unit definition of living is: under 50 DEG C, pH4.8 condition, 1 minute hydrolysis substrate CMC-Na produces the required enzyme amount of 1 μ g reducing sugar (with glucose meter) and is defined as 1 Ge Meihuo unit.
The cellulase of described step (2): mainly formed by circumscribed beta-glucanase, Endo-β-glucanase and beta-glucosidase etc.
The polygalacturonase of described step (2) is polygalacturonic acid lytic enzyme in essence.
The resin absorption of described step (3) is to select DA201 type styrene type polarity interpolymer macroporous resin adsorption.
The log dried firewood of described step (4) is the bavin of the self-produced trees felling gained of Dayao Mountain; Also can be the dry waste residue of Ocimum basilicum L. after extracting effective components.
The present invention, on the basis of Ocimum basilicum L. Manufacturing Techniques among the people in set ancient times, organically combines modern forward position extraction process, and the effective constituent that obtains Ocimum basilicum L. has following features:
1) the present invention has increased ripening before enforcement enzymolysis step, and its effect is to forge native mud pathogenic dryness, makes xenthophylls transfer lutein ester to, increases fragrance.It is high that this step has solved xenthophylls in new shade plant Ocimum basilicum L. herb, causes the problem that bioavailability is lower;
2) the present invention deposits characteristic according to the effective constituent of Ocimum basilicum L. in containing of plant, creatively utilize the enzymolysis process in modern cutting edge technology, first the cell walls of plant and intercellular pectin substance are destroyed, allow in Ocimum basilicum L. the preferential extract of aqueous soluble active constituent out.Destroy the Ocimum basilicum L. slag of intercellular pectin substance simultaneously, with ancient times folk art fire-cure, be in fact slaking or be oxidized oxygen containing terpenoid, more 9 to produce, 12-18-diolefinic acid, margaric acid, pohytol, butyl phthalate, the undecyl aldehyde etc.; And supercritical fluid extraction technique is not stopped up, and extracting and separating is easier.Practical data proves, can improve extract yield and reach more than 2 times, therefore enzymolysis process and ancient times folk art supercritical fluid extraction technique has been played to the technique effect of synergy;
3) the present invention utilizes supercritical CO
2extraction step extraction essential oil, has fragrance and approaches nature most, and essential oil extracting section thing extracts completely: 50 DEG C of following temperature extract, and do not destroy the advantage of aroma component;
4) technique of the present invention is extracted the effective constituent of Ocimum basilicum L., and yield is high, on average can reach more than 6%, and compared with Chinese invention patent application publication number, CNl414077A utilizes separately supercritical CO
2extraction yield is high more than 5 times.
Embodiment
The present invention will be described to enumerate a part of specific embodiment below, is necessary to be pointed out that at this following specific embodiment, only for the present invention is described further, does not represent limiting the scope of the invention.Some nonessential amendments that other people make according to the present invention and adjustment still belong to protection scope of the present invention.
Implement time of the present invention concrete, technical process be Ocimum basilicum L. taking fresh collection as raw material, through smoke → supercritical fluid extraction of ripening → enzymolysis → resin absorption → Ocimum basilicum L. log essential oil, obtain aqueous soluble active constituent extract and essential oil extracting section thing.
Embodiment 1
1) extraction of effective constituent from Ocimum basilicum L.
600 kilograms of the Ocimum basilicum L.s of fresh collection, wash away sand, and after wind drains, ageing 3-4 hour, to forge native mud pathogenic dryness; Be cut into after long broken section of 30-50mm with 200 kg water immersion 2 hours.Then be placed in stainless steel vessel, add cellulase and polygalacturonase and extract 4 hours.Filter, collect the effective component extracts in parting liquid liquid, obtain 70 kg water dissolubility effective component extracts products.Ocimum basilicum L. slag after enzymolysis is smoked to 6-8 hour with log, or smoke 10-12 hour with the dry waste residue of Ocimum basilicum L.; Hui Run, obtains 85 kilograms of Ocimum basilicum L. slags that fire-cure; By the Ocimum basilicum L. slag fire-cureing, adopt supercritical CO
2fluid is that extraction solvent extracts, and extracting solution separates and obtains essential oil extracting section thing through high speed centrifugation again; Supercritical CO
2extractant condition is: extracting pressure is 20MPa, and temperature is 42 DEG C, and the time is 3h, and flow is 300L/h; The condition of centrifugation is: rotating speed 6000-7000 rev/min, and temperature is normal temperature; Collect extract, obtain the essential oil portioned product of 6.95 kilograms of yellow transparent.Supercritical CO
2ocimum basilicum L. waste residue after extraction, dries.Can do Ocimum basilicum L. slag smokes and uses bavin.
2) analysis of Ocimum basilicum L. effective constituent
Adopt thin layer chromatography scanning, to make the thick thin layer plate that contains 0.5% CMC-Na (Xylo-Mucine) silica gel of 0.5mm by oneself, taking trichloromethane-methanol-water (mass ratio 80: 35: 10) as launching medicament, dual wavelength reflection method zigzag scanning, λ
s=260mm, λ
r=260mm, SX=3, slit 0.4mm × 0.4mm, the average extract yield 12.7% of Ocimum basilicum L. aqueous soluble active constituent triterpenoid saponin, the average extract yield 8.1% of the oxygen containing terpenoid essential oil of Ocimum basilicum L..
Embodiment 2
1) extraction of effective constituent from Ocimum basilicum L.
500 kilograms of the Ocimum basilicum L.s of fresh collection, wash away sand, and after wind drains, ageing 4 hours, to forge native mud pathogenic dryness; Be cut into after 30mm grows broken section and use double centner water soaking 2 hours.Then be placed in stainless steel vessel, add cellulase and polygalacturonase and extract 4 hours.Filter, collect the effective component extracts in parting liquid liquid, obtain 61 kg water dissolubility effective component extracts products.Ocimum basilicum L. slag after enzymolysis is smoked to 6-8 hour with log, or smoke 10-12 hour with the dry waste residue of Ocimum basilicum L.; Hui Run, obtains 72 kilograms of Ocimum basilicum L. slags that fire-cure; By the Ocimum basilicum L. slag fire-cureing, as polar solvent entrainment agent, adopt supercritical CO with ethanol
2fluid is that extraction solvent extracts, and extracting solution separates and obtains essential oil extracting section thing through high speed centrifugation again; Supercritical CO
2extractant condition is: extracting pressure is 18MPa, and temperature is 41 DEG C, and the time is 3h, and flow is 250L/h; The condition of centrifugation is: rotating speed 6000-7000 rev/min, and temperature is normal temperature; Collect extract, obtain the essential oil portioned product of 6.84 kilograms of yellow transparent.Supercritical CO
2ocimum basilicum L. waste residue after extraction, dries, and can do Ocimum basilicum L. slag and smoke and use bavin.
2) analysis of Ocimum basilicum L. effective constituent
Adopt thin layer chromatography scanning, to make the thick thin layer plate that contains 0.5% CMC-Na (Xylo-Mucine) silica gel of 0.5mm by oneself, taking trichloromethane-methanol-water (mass ratio 80: 35: 10) as launching medicament, dual wavelength reflection method zigzag scanning, λ
s=260mm, λ
r=260mm, SX=3, slit 0.4mm × 0.4mm, the average extract yield 12.7% of Ocimum basilicum L. aqueous soluble active constituent triterpenoid saponin, the average extract yield 9.4% of the oxygen containing terpenoid essential oil of Ocimum basilicum L..Illustrate at supercritical CO
2in extraction process step, add polar solvent entrainment agent can significantly improve extract yield.
Claims (5)
1. the method for an extracting effective components from Ocimum basilicum L., it is characterized in that: taking the Ocimum basilicum L. of fresh collection as raw material, through smoke → supercritical fluid extraction of ripening → enzymolysis → resin absorption → Ocimum basilicum L. log essential oil, obtain aqueous soluble active constituent extract and essential oil extracting section thing; Specifically comprise the steps:
(1) ripening
Gather fresh Ocimum basilicum L. herb, wash away sand, after draining, ageing 3-4 hour, to forge native mud pathogenic dryness;
(2) enzymolysis
Forge the herb after native mud pathogenic dryness, being cut into long broken section of 30-50mm puts in stainless steel vessel, till adding deionized water to alfalfa section and being just immersed in water, add 1000-1500U cellulase and 600-800U polygalacturonase by every kilogram of alfalfa section, with hydrochloric acid adjust pH to 4. 8, fully stir, 42 DEG C of water bath with thermostatic control 4 h, let cool to room temperature, extremely neutral with 0.2mol/L sodium hydroxide solution adjust pH, filtering separation;
(3) resin absorption
Step (2) parting liquid resin absorption; Resin after washing absorption until effluent liquid approach colourless, then the alcoholic solution wash-out that is 50-100% by concentration, collection elutriant, drying, obtains the Ocimum basilicum L. aqueous soluble active constituent extract of saponin content >=10%, this extract is specially triterpenoid saponin;
(4) Ocimum basilicum L. log is smoked
The Ocimum basilicum L. slag that step (2) separates, rinses through water, drains; Cut that to get on mountain bamboo several, length 3-5m, the two halves of riving, wash in a pan empty; The Ocimum basilicum L. slag draining is evenly laid in and is washed in a pan in half empty sheet bamboo, stay a little space ventilative, cover other half sheet bamboo, fix as bamboo folder cylinder; Each bamboo folder cylinder is controlled and separated slag loading level is 5kg; The bamboo folder cylinder that presss from both sides full Ocimum basilicum L. slag is emitted on bamboo bed, under bamboo bed, is paved with log dried firewood and lights a fire and fire-cure, while fire-cureing, will often stir the position of bamboo folder cylinder, make it uniform drying; The time of fire-cureing will be seen firepower size and determine, generally in 6-8 hour, is advisable; After fire-cureing, can take out the Ocimum basilicum L. slag fire-cureing and be placed on the ground, spray a little moisture, good with straw mulch, moisture is dissipated, after dry;
(5) supercritical fluid extraction essential oil
By the Ocimum basilicum L. slag fire-cureing, adopt supercritical CO
2fluid is that extraction solvent extracts, and extracting solution separates and obtains essential oil extracting section thing through high speed centrifugation again; Supercritical CO
2extractant condition is: extracting pressure is 17-20MPa, and temperature is 40-42 DEG C, and the time is 2-3h, and flow is 250-300L/h; The condition of centrifugation is: rotating speed 6000-7000 rev/min, temperature is normal temperature.
According to claim 1 from Ocimum basilicum L. the method for extracting effective components, it is characterized in that; The cellulase of described step (2): mainly formed by circumscribed beta-glucanase, Endo-β-glucanase and beta-glucosidase; The polygalacturonase of described step (2) is polygalacturonic acid lytic enzyme in essence.
According to claim 1 from Ocimum basilicum L. the method for extracting effective components, it is characterized in that; The resin absorption of described step (3) is to select DA201 type styrene type polarity interpolymer macroporous resin adsorption.
According to claim 1 from Ocimum basilicum L. the method for extracting effective components, it is characterized in that; The log dried firewood of described step (4) is the bavin of the self-produced trees felling gained of Dayao Mountain.
According to claim 1 from Ocimum basilicum L. the method for extracting effective components, it is characterized in that; The log dried firewood of described step (4) is the dry waste residue of Ocimum basilicum L. after extracting effective components.
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| KR101781635B1 (en) * | 2015-07-31 | 2017-09-25 | 남부대학교산학협력단 | A composition for control of melanose comprising the extract of lysimachiae foenum-graeci herba as an effective component, microcapsule comprising of the same and a control method of melanose using the same |
| CN105861563B (en) * | 2016-06-03 | 2019-03-19 | 湖北中烟工业有限责任公司 | A kind of complex art prepares the method and its application of Guanling perfume (or spice) banana aldehyde |
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| CN1414077A (en) * | 2002-09-27 | 2003-04-30 | 广西壮族自治区中国科学院广西植物研究所 | Extraction method of high quality lingxiang grass purified oil |
| CN101037465B (en) * | 2006-03-15 | 2010-04-21 | 北京同仁堂股份有限公司 | Saponins and preparation method and application |
| CN100491393C (en) * | 2006-03-15 | 2009-05-27 | 北京同仁堂股份有限公司 | Steroid saponins and preparation method and application |
| CN101700275A (en) * | 2009-11-17 | 2010-05-05 | 广东药学院 | Pharmaceutical preparation containing axillary choerospondias fruit and preparation method and application thereof |
| CN101875685A (en) * | 2010-06-25 | 2010-11-03 | 广西师范大学 | Lysimachia foenum-graecum hance saponin extract having more than or equal to 30 percent of saponin and extraction process thereof |
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