CN103272065B - Method for extracting volatile oil from agilawood seeds, and usage and application of volatile oil - Google Patents
Method for extracting volatile oil from agilawood seeds, and usage and application of volatile oil Download PDFInfo
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Abstract
The invention discloses a method for extracting a volatile oil from agilawood seeds, and a usage and an application of the volatile oil. In the method disclosed by the invention, a supercritical CO2 fluid extraction method is adopted to extract volatile components from aquilaria sinensis seeds; and a DPPH method is used to study anti-oxidation capability of effective components of the volatile components. With the method, squalene with relatively high content is detected in the aquilaria sinensis seeds for the first time; and the volatile oil has a certain anti-oxidation capability, so that scientific basis is provided for further studying active components of the aquilaria sinensis seeds development and utilization The extraction method is simple in process flow, high in extraction yield and low in extraction temperature, and can prevent degradation of heat-sensitive components. The method fully develops rich resources of the agilawood rare medicinal material, and makes the best use of the agilawood. With the method, effective components in the agilawood seeds are extracted simply and effectively, thereby developing novel raw materials and extraction process for the squalene.
Description
[technical field]
The present invention relates to a kind of extracting method of Lignum Aquilariae Resinatum seed extract, particularly a kind of method extracting volatile oil from Lignum Aquilariae Resinatum seed, the invention still further relates to purposes and the application of this Lignum Aquilariae Resinatum seed extract volatile oil, belong to biological medicine, health care technology field.
[background technology]
The resiniferous timber of Lignum Aquilariae Resinatum Aquilaria sinensis (lour.) Gilgl. is called Lignum Aquilariae Resinatum, is also called Home-made occluder, Aquilaria sinensis (Lour.) Spreng. or Aquilaria sinensis, and Lignum Aquilariae Resinatum Ye Shi China produces unique plant resources of Lignum Aquilariae Resinatum medical material.The wild resource of Lignum Aquilariae Resinatum is endangered, has now been listed in national Precious, Rare, Endangered three-level protective plant and national secondary national key protected plant.Lignum Aquilariae Resinatum, acrid in the mouth, hardship, warm in nature, having effect of promoting the circulation of QI to relieve pain, warming middle-JIAO to arrest vomiting, helping inspiration to relieve asthma, be used for the treatment of the vexed pain of breast abdominal distention, gastrofrigid vomiting singultus, QI rising in reverse order of suffering from a deficiency of the kidney dyspnea with rapid respiration, is the traditional precious Chinese medicine of China.
Lignum Aquilariae Resinatum is high megaphanerophyte, poor growth, and the Edgeworthia chrysantha Lindl. cycle is long, needs 5 years above diameters of a cross-section of a tree trunk 1.3 meters above the ground of the age of tree to reach 15cm and just can carry out man perfume (or spice).And Lignum Aquilariae Resinatum can be yielded positive results in plantation for 3 years, a large amount of seed of annual generation, only have small part seed at present for nursery, its affluent resources are also effectively developed.Research finds, it is active that Lignum Aquilariae Resinatum seed has certain methicillin-resistant staphylococcus aureus resistance (MRSA), and have not been reported the exploitation of its other medicinal parts and the comprehensive utilization of resource.
Squalene has another name called zamene, squalene, Squalene, it is a kind of height unsaturated hydrocarbons compounds, found in black shark liver oil in 1906 by Japanese chemists Tsujimoto at first, Squalene is a kind of lipid non-saponifiable matter, also known as Squalene, have and improve superoxide dismutase (SOD) activity in body, enhancing human body immunity ability, improvement function, defying age, resisting fatigue, the different physiological roles such as antitumor, Squalene has extremely strong oxygen delivery capacity, can generate by anticancer, prevent cancerous cell from spreading and the leukopenia because of chemotherapy, also there is resisting fatigue simultaneously, anti-cardiovascular disease, anti-infective effect, it is a kind of avirulent marine biomaterial with the effect of preventing and curing diseases.Squalene distributes very wide in plant, but content is not high, and how lower than 5% of non-saponifiable matter in vegetable oil, only minority content is more, and as olive oil contains Squalene 150-700mg/100g, Testa oryzae oil content is 332mg/100g; Oleum Camelliae is the general name to oily substance in Semen Camelliae, also containing more Squalene in Oleum Camelliae.Because in deepwater shark liver, the content of Squalene is high, if content in armour shark-liver oil is 40%-74%, spinule shark, containing 49%-89%, therefore, mostly adopts deepwater shark liver oil to extract Squalene at present; But so produce Squalene in a large number, the disruption of ecological balance of deepwater shark can be caused.
Also have the method extracting Squalene on a small quantity from plant to announce in recent years, Chinese patent (CN201110027119.4) " taking Semen Camelliae as method and the Oleum Camelliae squalene soft capsules that raw material extracts Squalene " utilizes lixiviate, chromatographic isolation and room temperature extraction and several extracting method of supercritical extraction to combine the high-purity squalene extracting and be suitable for; Chinese patent (CN200910069670.8) " taking olive oil as the method that raw material extracts high-purity squalene " is that the separating technology combined with secondary molecules distillation and silica gel chromatography extracts high-purity squalene; The process that Chinese patent (CN201010190275.8) " extracts the method for Squalene " and utilizes molecular distillation to be combined with extraction, crystalline phase from plant oil deodorizing distillate, extracts high concentration Squalene; These methods all need multiple extracting method to be combined, and technique is more complicated all, introduce and have other chemical substances, more difficult control during operating procedure.
Containing abundant Squalene in Lignum Aquilariae Resinatum seed, being expected to exploitation becomes medicine, cosmetics and health food etc.
[summary of the invention]
Object of the present invention is just in order to make up blank and the deficiency of prior art, there is provided a kind of technological process simple, extraction efficiency is high, and extraction temperature is low, the method extracting volatile oil from Lignum Aquilariae Resinatum seed that heat-sensitive ingredients can be avoided to degrade, opens the development approach of new medicinal part in Lignum Aquilariae Resinatum seed.
Another object of the present invention is to provide purposes and the application of volatile oil in a kind of above-mentioned Lignum Aquilariae Resinatum seed.
The present invention to achieve these goals, by the following technical solutions:
From Lignum Aquilariae Resinatum seed, extract a method for volatile oil, it is characterized in that adopting supercritical carbon dioxide to extract.
A kind of method extracting volatile oil from Lignum Aquilariae Resinatum seed of the present invention, it comprises the following steps:
Get dry Lignum Aquilariae Resinatum seed, pulverized with pulverizer, be then placed in supercritical extraction reactor with the CO of 10-30L/h
2at 40 ~ 50 DEG C, under 18 ~ 25MPa condition, extract 1.5 ~ 2.5h, then fluid after extraction is carried out twice decompression separation and collect extract, obtain volatile oil.
A kind of method extracting volatile oil from Lignum Aquilariae Resinatum seed of the present invention, twice decompression separation wherein comprises the first separating still, pressure 5 ~ 7MPa, and temperature is 45 ~ 50 DEG C, and the second separating still, pressure 5 ~ 7MPa, temperature 45 ~ 50 DEG C.
The present invention is a kind of to extract in the method for volatile oil from Lignum Aquilariae Resinatum seed, and Lignum Aquilariae Resinatum seed crosses 20 ~ 40 mesh sieves after pulverizing.
The present invention is a kind of to extract in the method for volatile oil from Lignum Aquilariae Resinatum seed, water content≤10% of dry Lignum Aquilariae Resinatum seed
Effective ingredient in the volatile oil that the present invention extracts is Squalene.
The volatile oil that the present invention extracts from Lignum Aquilariae Resinatum seed is for removing the purposes of DPPH free radical.
The application preparation of the volatile oil that the present invention extracts from Lignum Aquilariae Resinatum seed is soft capsule.
When the application preparation of the volatile oil that the present invention extracts from Lignum Aquilariae Resinatum seed is soft capsule, this soft capsule is mixed with uniform liquid filling by volatile oil, vitamin E and soybean oil.
Adopt supercritical CO of the present invention
2volatile oil in fluid extraction Lignum Aquilariae Resinatum seed, has following advantage:
Technological process is simple, extraction efficiency is high, extraction temperature is low, the degraded of heat-sensitive ingredients can be avoided, the present invention detects the Squalene that content is higher first in the volatile oil of Lignum Aquilariae Resinatum seed, this volatile oil has certain oxidation resistance, for studying its active component further and exploitation provide scientific basis.
The present invention adopts the annual superfluous a large amount of seeds of Lignum Aquilariae Resinatum, fully develop the affluent resources of this valuable ingredient of Chinese medicine of Lignum Aquilariae Resinatum, accomplish and make the best use of everything, the present invention is extracted the effective ingredient in Lignum Aquilariae Resinatum seed simply and effectively, for Squalene opens new raw material and extraction process.
Squalene content in the volatile oil that the present invention extracts is high, and its content accounts for 41.345% of volatile ingredient, and other subsidiary component contents are lower, and the effectiveness not affecting Squalene composition plays, and is also easy to be separated.
The volatile oil that the present invention extracts has extremely strong oxygen delivery capacity, can generate by anticancer, prevents cancerous cell from spreading and the leukopenia because of chemotherapy, also has resisting fatigue, anti-cardiovascular disease, anti-infective effect simultaneously.Containing abundant Squalene in Lignum Aquilariae Resinatum seed, being expected to exploitation becomes medicine, cosmetics and health food etc.
Present invention process is simply single, easy to control, does not introduce new impurity in the volatile oil extracted, and other compositions original do not affect the use of main composition Squalene yet.
On the other hand, being found Lignum Aquilariae Resinatum fat content is 68.5%, far above the seed of other xylophytas, there are the potentiality as oil plant or biodiesel resource, the present invention does not effectively introduce again the volatile oil be extracted in Lignum Aquilariae Resinatum seed of new impurity, for the abundant development and utilization of Lignum Aquilariae Resinatum seed opens new way.
[accompanying drawing explanation]
Fig. 1 is Lignum Aquilariae Resinatum Volatile Oil from Seed GC-MS analyzing total ion flow;
Fig. 2 is the absorption spectrum curve figure of DPPH;
Fig. 3 is volatile oil sample solution trap curve chart over time;
Fig. 4 is the bar diagram of Lignum Aquilariae Resinatum Volatile Oil from Seed scavenging ability of DPPH free radical.
[detailed description of the invention]
Below in conjunction with detailed description of the invention, the invention will be further described:
One, instrument and medical material
Material: medical material is Lignum Aquilariae Resinatum Aquilaria sinensis (lour.) the Gilgl. seed of planting base, Lignum Aquilariae Resinatum mountain, Maoming, is accredited as certified products through author.
Instrument: gas phase-mass spectrometry chromatograph is the Agilent6890-5973N type of Agilent company of the U.S.; Supercritical extracting equipment is Jiangsu Huaan Science Research Instrument Co., Ltd HA121-50-05 model; Ultraviolet spectrophotometer is the UV-2550 model of Japanese Shimadzu instrument company.
Reagent: DPPH (1,1-Dipheyl-2-picrylhydrazyl radicle, 1,1-diphenyl-2-picryl hydrazine) Aladdin reagent (Shanghai) Co., Ltd.; Dehydrated alcohol Shantou Xilong Chemical Co., Ltd, is analytical pure.
Two, extracting method:
The present invention extracts Lignum Aquilariae Resinatum seed when waving oil, gets dry Lignum Aquilariae Resinatum seed, is crushed to 20 ~ 40 mesh sieves, and was then placed in supercritical CO with pulverizer
2with the flow of 20L/h in extraction kettle, at 40 ~ 50 DEG C, 1.5 ~ 2.5h is extracted under 18 ~ 25MPa condition, then respectively with first separating still pressure 5 ~ 7MPa, temperature 45 ~ 50 DEG C, second separating still pressure 5 ~ 7MPa, temperature 45 ~ 50 DEG C of collections have the yellow transparent grease of strong fragrance, i.e. volatile oil.
Three, chemical composition analysis
GC-MS analysis condition
Chromatographic condition:
Chromatographic column Agilent HP-5MS (0.25umx250umx30m), injector temperature is 250 DEG C, and carrier gas is He gas, post flow is 1.0ml/min, temperature programming condition: initial temperature 50 DEG C, rises to 220 DEG C with 4 DEG C/min, after rise to 300 DEG C with 10 DEG C/min.
Mass Spectrometry Conditions:
Normal pictures is tuning, and ion source temperature is 230 DEG C, and level Four bar temperature is 150 DEG C, and data acquisition scan pattern is full scan, solvent delay time 3min.
Gas chromatograph-mass spectrometer (GC-MS) (GC-MS) is adopted to be separated and content analysis Lignum Aquilariae Resinatum seed ethereal oil, by above-mentioned analysis condition, its gas chromatography-mass spectrum (GC-MS) total ion current figure as shown in Figure 1, vertical coordinate represents abundance, abscissa representing time, gained chromatograph and Information in Mass Spectra carry out retrieval and manual retrieval, contrast and parsing automatically through NIST2011 standard spectrum storehouse, chemical composition in qualification Lignum Aquilariae Resinatum volatile oil, determines the mass fraction of each composition by area normalization method.
9 main compound in this experimental identification Lignum Aquilariae Resinatum volatile oil, composition majority is fatty acid, what content was higher has Squalene squalene (41.345%), ethyl oleate ethyl oleate (32.233%) and n-hexadecanoic acid (16.708%) etc., and concrete analysis the results are shown in Table 1:
Table 1: Lignum Aquilariae Resinatum Analysis of The Essential Oil result C
14
Four, the mensuration of volatile oil oxidation resistance
The preparation of DPPH solution:
Precision takes in 0.0198g DPPH to 50ml volumetric flask, is settled to graduation mark, as storing solution with dehydrated alcohol.Used time precision pipettes in storing solution 10ml to 100ml volumetric flask, obtains the DPPH working solution of 0.1mmol/L with dehydrated alcohol standardize solution.
The preparation of sample solution:
With dehydrated alcohol, Lignum Aquilariae Resinatum Volatile Oil from Seed is mixed with low concentration group and high concentration group, low concentration group respectively: 2,4,6,8,10mg/mL sample solution; High concentration components is not: 20,40,60,80,100mg/mL sample solution.
The determination of maximum absorption wavelength:
DPPH solution is made to carry out spectral scan in 280 ~ 700nm range of wavelengths, can find that DPPH all has strong absworption peak at 326nm and 516nm place, after adding sample solution, these two peak values all have reduction, and the peak value at 516nm wavelength place reduces comparatively remarkable, as shown in Figure 2, therefore, 516nm is chosen as mensuration wavelength.
The determination in response time:
Getting 1ml20mg/mL sample solution joins in 7ml DPPH solution, and mix homogeneously surveys its absorbance (A value), until A value is basicly stable every 5min.Solution A value over time rule as shown in Figure 3, as seen from Figure 3, in the 20min adding sample, absorbance declines very fast, and along with the increase of time, the downward trend of absorbance tends towards stability gradually, react after 40min and substantially reach stable, therefore select 40min as the response time.The antioxidation of fixation response time measures:
The sample solution pipetting 2ml variable concentrations respectively adds in the DPPH solution of 4ml, mix homogeneously, and room temperature places 40min, measures its absorbance (A), parallel assay 3 times at maximum absorption wavelength 516nm place; To add the DPPH solution of 2ml dehydrated alcohol as blank (A
0).Last according to each concentration samples of following formulae discovery to the clearance rate of DPPH free radical: clearance rate (ρ)=1-A/A
0× 100%.
DPPH molecule can generate stable containing nitrogen free radical in the solution, and solution colour shows purple, when it and free radical scavenger effect, can generate colourless product, solution colour is shoaled.Different antioxidants is different to the clearance rate of free radical and elimination effect, and clearance rate can reflect the scavenging action to free radical, and clearance rate is larger, and antioxidant activity is stronger, otherwise then more weak.
The contrast of the Lignum Aquilariae Resinatum volatile oil scavenging ability of DPPH free radical of variable concentrations as shown in Figure 4, result shows, the volatile oil of Lignum Aquilariae Resinatum seed has certain Scavenging activity to DPPH free radical, high concentration group clearance rate is apparently higher than low concentration group, and along with the increase of concentration, clearance rate increases gradually, when concentration is 80mg/mL, clearance rate reaches 82.8%, and afterwards along with concentration increases, clearance rate increases not obvious.
Five, be several specific embodiment of the present invention below
Embodiment 1:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 40 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 40 DEG C, 1.5h is extracted under 25MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 6MPa, temperature 50 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 132g, extraction yield is 44%, and the extraction ratio of Squalene is 16.72%.
Embodiment 2:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 30 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 40 DEG C, 2h is extracted under 18MPa condition, then respectively at the first separating still (pressure 6MPa, temperature 48 DEG C) and the second separating still (pressure 5MPa, temperature 48 DEG C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 117g, extraction yield is 39%, and the extraction ratio of Squalene is 14.04%.
Embodiment 3:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 20 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 40 DEG C, 2.5h is extracted under 21MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 45 C) and the second separating still (pressure 5MPa, temperature 45 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 147g, extraction yield is 49%, and the extraction ratio of Squalene is 19.11%.
Embodiment 4:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 20 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 45 DEG C, 1.5h is extracted under 18MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 6MPa, temperature 48 DEG C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 105g, extraction yield is 35%, and the extraction ratio of Squalene is 14.7%.
Embodiment 5:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 40 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 45 DEG C, 2h is extracted under 21MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 5MPa, temperature 45 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 189g, extraction yield is 63%, and the extraction ratio of Squalene is 25.83%.
Embodiment 6:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 30 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 45 DEG C, 2.5h is extracted under 25MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 5MPa, temperature 45 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 138g, extraction yield is 46%, and the extraction ratio of Squalene is 19.32%.
Embodiment 7:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 30 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 50 DEG C, 1.5h is extracted under 21MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 7MPa, temperature 50 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 162g, extraction yield is 54%, and the extraction ratio of Squalene is 20.52%.
Embodiment 8:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 20 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 50 DEG C, 2h is extracted under 25MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 5MPa, temperature 45 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 138g, extraction yield is 46%, and the extraction ratio of Squalene is 16.56%.
Embodiment 9:
Get the dry Lignum Aquilariae Resinatum seed of water content≤10%, be crushed to 40 mesh sieves with pulverizer, accurately take 300g and be then placed in extraction kettle with the CO of 20L/h
2at 50 DEG C, 2.5h is extracted under 18MPa condition, then respectively at the first separating still (pressure 7MPa, temperature 50 C) and the second separating still (pressure 5MPa, temperature 45 C) in decompression collect the yellow transparent grease of strong fragrance, i.e. volatile oil 144g, extraction yield is 48%, and the extraction ratio of Squalene is 17.76%.
From above-described embodiment parameter and interpretation of result, in the influence factor of extraction yield in extracting method of the present invention, extracting pressure > raw meal particle size > extraction temperature > extraction time.
And the extraction ratio extracting Squalene from other raw materials the highest be just 2.09%.The present invention is compared with existing extraction Squalene method, and extraction ratio is high, and technique is simple, safe and reliable.
Claims (3)
1. a purposes for the volatile oil extracted from Lignum Aquilariae Resinatum seed with supercritical carbon dioxide, is characterized in that, for removing DPPH free radical, wherein the effective ingredient of Lignum Aquilariae Resinatum Volatile Oil from Seed is Squalene;
The preparation method of described Lignum Aquilariae Resinatum Volatile Oil from Seed is:
Get dry Lignum Aquilariae Resinatum seed, be crushed to 20 ~ 40 mesh sieves with pulverizer, and be then placed in supercritical CO
2with the flow of 20L/h in extraction kettle, at 40 ~ 50 DEG C, 1.5 ~ 2.5h is extracted under 18 ~ 25MPa condition, then respectively with first separating still pressure 5 ~ 7MPa, temperature 45 ~ 50 DEG C, second separating still pressure 5 ~ 7MPa, temperature 45 ~ 50 DEG C of collections have the yellow transparent grease of strong fragrance, i.e. volatile oil.
2. the purposes of volatile oil that extracts from Lignum Aquilariae Resinatum seed of a kind of supercritical carbon dioxide according to claim 1, is characterized in that the volatile oil that described supercritical carbon dioxide extracts from Lignum Aquilariae Resinatum seed is soft capsule for removing the application preparation of DPPH free radical.
3. the purposes of volatile oil that extracts from Lignum Aquilariae Resinatum seed of supercritical carbon dioxide according to claim 2, is characterized in that described soft capsule is mixed with uniform liquid by volatile oil, vitamin E and soybean oil and fills.
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| CN103981020A (en) * | 2014-05-16 | 2014-08-13 | 吴江市英力达塑料包装有限公司 | Extraction-separation method of eaglewood oil |
| CN104940050A (en) * | 2015-05-07 | 2015-09-30 | 孙广娥 | Use of agilawood volatile oil stoste in preparation of skin-care product |
| CN108601808A (en) * | 2015-11-16 | 2018-09-28 | 澄霖农业生技股份有限公司 | It is a kind of that human body is contributed to repair the agalloch eaglewood food that diabetes cause renal lesions |
| CN107778724B (en) * | 2017-11-17 | 2020-04-21 | 广州中医药大学 | Agilawood seed extract and preparation method and application thereof |
| TWI726645B (en) * | 2020-03-04 | 2021-05-01 | 謝慧萍 | Use of agarwood tree extract to prepare skin repair composition |
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| CN101129316A (en) * | 2007-08-30 | 2008-02-27 | 中国热带农业科学院热带生物技术研究所 | Ligaloes perfumes and method of producing the same |
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