CN113462475A - Extraction method and application of yulan magnolia leaf cell water and yulan magnolia leaf essential oil - Google Patents
Extraction method and application of yulan magnolia leaf cell water and yulan magnolia leaf essential oil Download PDFInfo
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/022—Refining
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
- A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/025—Recovery by solvent extraction
-
- C—CHEMISTRY; METALLURGY
- C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
- C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
- C11B9/00—Essential oils; Perfumes
- C11B9/02—Recovery or refining of essential oils from raw materials
- C11B9/027—Recovery of volatiles by distillation or stripping
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
Abstract
The invention discloses a method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil, which mainly utilizes a low-temperature-vacuum (reduced pressure) extraction technology to primarily extract and separate a part of liquid (yulan magnolia leaf cell water and yulan magnolia leaf essential oil) in yulan magnolia leaves in a short time, a certain amount of cellulase and pectinase are added into the liquid, the liquid is reversely poured into the primarily extracted yulan magnolia leaf residues, the low-temperature-reduced pressure extraction is performed for a certain time again, an extracting solution is collected, and after standing and layering, the yulan magnolia leaf cell water and the fragrant yulan magnolia leaf essential oil which are rich in active ingredients, fragrant and high in safety are obtained. The extraction method of the invention can control the chemical composition in cell water and essential oil.
Description
Technical Field
The invention relates to the technical field of agricultural product treatment, in particular to an extraction method and application of yulan magnolia leaf cell water and yulan magnolia leaf essential oil.
Background
Magnolia denudataDesr is a deciduous tree of Magnoliaceae, or Magnolia genus. The Magnolia denudata leaf has effects of clearing heat, promoting urination, relieving cough, eliminating phlegm, and treating urinary system infection, dysuresia, and bronchitis. The magnolia leaves contain rich vitamins, amino acids and various trace elements, and have the effects of expelling wind and clearing away cold, and promoting qi and regulating lung. The magnolia leaves are also a traditional Chinese medicine capable of relieving cough and asthma, can diminish inflammation, can also dilute sputum, can dilate trachea, and can quickly improve adverse symptoms of cough, asthma, excessive phlegm and the like. Although the magnolia leaves have lower medicinal value compared with magnolia, the magnolia leaves have wide sources and low price, and have great development prospect.
The main extraction methods of the plants at the present stage include a distillation method, an extraction method and a supercritical carbon dioxide extraction method. The distillation method has long time consumption and large energy consumption; the leaching method needs additional solvent, so that the storage and use of the extract are limited; the supercritical carbon dioxide extraction method has high requirements on equipment, expensive equipment and high extraction cost, and is not suitable for industrial production.
Enzymatic methods are also commonly used in the extraction of plant cell sap. The Chinese patent application CN109730948A discloses a method for preparing peony flower cell water by combining an ultrasonic low-temperature rotary steaming method and an enzyme method, which comprises the following steps: firstly, squeezing to obtain juice and residue 1, then carrying out rotary evaporation on the residue 1 to obtain cell water 1 and residue 2, and finally carrying out enzymatic hydrolysis on the residue 2 and then carrying out rotary evaporation to obtain cell water 2. And mixing the juice, the cell water 1 and the cell water 2 to obtain the high-yield peony cell water. The method has high extraction efficiency, but has the following defects: (1) the squeezing method is adopted and then mixed with the liquid obtained by the vacuum extraction method, so that polysaccharide, pigment and pungent smell are brought in, and the problems of corrosion prevention and decoloration are caused; (2) the method solves the problem of corrosion resistance by being matched with other plants for distillation at the later stage of the process, but easily changes the original water content and smell of the peony cells, and the quality cannot be controlled in the later-stage production.
Plant cells are rich in flavonoids, and the flavones are easily oxidized to turn yellow, so that the quality of the essential oil is reduced. In the prior art, flavone and polysaccharide are mainly removed through post-treatment.
Disclosure of Invention
The invention aims to provide a method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil, which has high extraction speed and can control the chemical compositions of the yulan magnolia leaf cell water and the yulan magnolia leaf essential oil.
The invention is realized by the following technical scheme:
a method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil comprises the following steps: the method comprises the steps of adding no solvent, primarily extracting magnolia leaves at the temperature of 30-60 ℃ and under the pressure of-60 kPa to-101 kPa, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam in the extraction process, collecting liquid through condensation, extracting for 1.5-3 hours to obtain primary extracted liquid and primary extracted magnolia leaf residue, adding 0.2-0.4% of cellulase and 0-0.1% of pectinase based on the total weight of the primary magnolia leaves into the primary extracted liquid, then adding the primary extracted liquid into the primary extracted magnolia leaf residue, extracting for 3-7 hours again under the conditions of 30-45 ℃ and under the pressure of-60 kPa to-101 kPa, and layering the collected liquid after extraction to obtain the magnolia leaf cell water and the magnolia leaf essential oil.
The primary extraction time is one of key parameters, if the time is too short, the extracted yulan magnolia leaf cell water is too little, and the cell water is difficult to infiltrate the surface of the yulan magnolia leaf after the enzyme is added, so that the enzymolysis can not be normally carried out. If the primary extraction time is too long, cell water and essential oil flow out too much, so that the subsequent extraction efficiency is reduced, the enzymolysis time is also increased, and the risk of excessive enzymolysis is brought. The method has the following advantages that a certain amount of enzyme is added into the primary extraction liquid, and the primary extraction liquid is put into the container again to extract the residues. Firstly, the initial liquid surface tension is low, and the permeability is good; secondly, the pH of the primary extraction liquid is 3-7, and the pH does not need to be additionally adjusted, so that the enzyme activity is favorably improved; thirdly, the enzymolysis can accelerate the wall breaking; fourthly, low-temperature vacuum technology extraction. Through the synergy of the four effects, the enzymolysis speed can be controlled at a lower temperature (35-45 ℃) to accelerate the cell sap outflow speed. About the first 1 hour in the re-extraction step, the primary extraction liquid poured back into the container can be steamed off, the enzymolysis speed is accelerated, the enzymolysis duration is shortened (at the moment, the amount of the primary extraction liquid is very important, the enzymolysis time is prolonged, and the enzymolysis time is shortened), and the problems that the traditional enzymolysis method needs to add a large amount of water to dilute the cell sap and the pungent smell caused by excessive enzymolysis is avoided.
Regarding the permeability of the initially extracted liquid, it was found through experiments that when the yulan magnolia leaf residue is extracted using the cell sap as the solvent of the solvent method, a large amount of macromolecular substances such as flavone and polysaccharide and volatile active ingredients can be carried out. Compared with pure water as a solvent, the cell water as the solvent can extract more active ingredients such as flavone and polysaccharide.
In addition, macromolecular substances such as flavone and polysaccharide cannot be extracted by the method, and the stability of the cell water and the essential oil is further improved.
Preferably, the primary extraction temperature is 35 to 50 ℃ and the pressure is-80 to-101 kPa. In the preferable condition range, the water in the plant cells reaches a boiling state, the extraction speed is high, and the extraction temperature is close to room temperature, so that the water activity of the volatilized cells can be kept, and the separated primary extraction liquid has good permeability.
Preferably, the addition amount of the cellulase is 0.25-0.35%, and the addition amount of the pectinase is 0.01-0.06%. The enzyme is macromolecular protein, can be adsorbed on the surface of magnolia leaf cells within the addition range, is rarely dissolved in water and essential oil, is difficult to volatilize under the low-temperature vacuum condition, and does not need subsequent treatment.
The magnolia leaves can be cut into the width of 1 mm-10 mm and then extracted, and simultaneously, the mixture is stirred in the extraction process, and the stirring speed is 1-150 r/min; can accelerate the extraction speed and is beneficial to fully extracting cell water and essential oil.
Condensing in the collection process, wherein the temperature is-10-8 ℃.
The invention adopts fresh magnolia leaves, namely newly picked magnolia leaves which are full of moisture, have no mildew and have no rot; generally, fresh leaves are green and full, not withered and yellow, and not baked or dried in the sun.
The magnolia leaves are preferably old magnolia leaves, and the content of active ingredients in the old magnolia leaves is higher than that in the tender magnolia leaves. Old leaves are mature leaves, the length of the old leaves of the magnolia is 10-18 cm, and the width of the old leaves of the magnolia is 6-12 cm. Generally, tender leaves of magnolia are light green, and the leaves are soft; the old magnolia leaves are dark green and hard in leaf quality.
The magnolia leaf cell water and the magnolia leaf essential oil obtained by the extraction method are green and natural, have no other solvents or additives, do not contain heavy metals, have fresh and pleasant aroma, and can be applied to skin care products, foods and health care products.
Compared with the prior art, the invention has the following beneficial effects:
the method firstly extracts a certain amount of cell sap by a low-temperature vacuum extraction technology, and then extracts the magnolia leaf residues again after adding enzyme by utilizing the high permeability of the primary extracted cell sap. Can accelerate the extraction efficiency of cell water and essential oil, avoids the problem of excessive enzymolysis brought by the traditional enzymolysis method, can obtain more cell sap (especially more essential oil) compared with the low-temperature vacuum extraction technology, and has more active ingredients.
The magnolia leaf cell water obtained by the method is clear and transparent, has high purity, contains more than 50 volatile active ingredients, has fresh and pleasant fragrance and high safety to human bodies, and can be applied to the fields of skin care products, foods, health care products and the like. The magnolia leaf essential oil obtained by the method does not contain polysaccharide and flavone, and has good stability.
Drawings
FIG. 1 is the water safety test data of the leaf cells of Yulan magnolia of example 1.
Detailed Description
The present invention will be described in detail with reference to specific examples. The following examples will assist those skilled in the art in further understanding the invention, but are not intended to limit the invention in any way. It should be noted that variations and modifications can be made by persons skilled in the art without departing from the spirit of the invention. All falling within the scope of the present invention.
The magnolia leaves used in the examples and comparative examples are fresh, non-mildew, old magnolia leaves from Guangxi. Cleaning, draining, cutting into pieces with the width of 3-5 mm, and carrying out extraction experiments.
Example 1:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding no solvent, primarily extracting under the conditions of 40 ℃, pressure of minus 101kPa and 45 rpm stirring, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam, condensing and collecting (the condensing temperature is minus 5 ℃), extracting for 2 hours, separating to obtain primary extraction liquid and primary extraction residue, adding 0.32 percent of cellulase and 0.04 percent of pectinase which are based on the total weight of the original magnolia leaves into the primary extraction liquid, adding the primary extraction liquid into the primary extraction residue, extracting the magnolia leaf cell water and the magnolia leaf essential oil again under the conditions of 40 ℃, pressure of minus 101kPa and 45 rpm stirring, standing for 6 hours, and layering the collected liquid to obtain 36.7kg of the magnolia leaf cell water and 6.3g of the magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow, and has fresh and pleasant fragrance.
Example 2:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding no solvent, primarily extracting under the conditions of 65 ℃, pressure of-60 kPa and 120 rpm stirring, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam, condensing and collecting (the condensing temperature is-5 ℃), extracting for 1.5 hours, separating to obtain primary extraction liquid and primary extraction residues, adding 0.40 percent of cellulase and 0.01 percent of pectinase which are based on the total weight of the original magnolia leaves into the primary extraction liquid, adding the primary extraction liquid into the primary extraction residues, extracting the magnolia leaf cell water and the magnolia leaf essential oil again under the conditions of 40 ℃, pressure of-90 kPa and 120 rpm stirring, finishing extraction for 5 hours, and standing and layering the collected liquid to obtain 35.4kg of the magnolia leaf cell water and 5.3g of the magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow, and has fresh and pleasant fragrance.
Example 3:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding no solvent, primarily extracting under the conditions of 50 ℃, pressure of-90 kPa and 60 rpm stirring, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam, condensing and collecting (the condensing temperature is-5 ℃), extracting for 1.8 hours, separating to obtain primary extraction liquid and primary extraction residues, adding 0.32 percent of cellulase and 0.04 percent of pectinase which are based on the total weight of the original magnolia leaves into the primary extraction liquid, adding the primary extraction liquid into the primary extraction residues, extracting the magnolia leaf cell water and the magnolia leaf essential oil again under the conditions of 45 ℃, pressure of-90 kPa and 60 rpm stirring, finishing extraction for 6 hours, and standing and layering the collected liquid to obtain 36.3kg of the magnolia leaf cell water and 6.1g of the magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow, and has fresh and pleasant fragrance.
Example 4:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding no solvent, primarily extracting under the conditions of 30 ℃, pressure of-101 kPa and 45 rpm stirring, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam, condensing and collecting (the condensing temperature is-5 ℃), extracting for 2.5 hours, separating to obtain primary extraction liquid and primary extraction residues, adding 0.2 percent of cellulase and 0.06 percent of pectinase based on the total weight of the original magnolia leaves into the primary extraction liquid, adding the primary extraction liquid into the primary extraction residues, extracting the magnolia leaf cell water and the magnolia leaf essential oil again under the conditions of 35 ℃, pressure of-101 kPa and 45 rpm stirring, finishing extraction for 3.5 hours, and standing and layering the collected liquid to obtain 35.0kg of the magnolia leaf cell water and 5.3g of the magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow, and has fresh and pleasant fragrance.
Example 5:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding no solvent, primarily extracting under the conditions of 45 ℃, pressure of-90 kPa and 80 rpm stirring, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam, condensing and collecting (the condensing temperature is-5 ℃), extracting for 2 hours, separating to obtain primary extraction liquid and primary extraction residue, adding 0.28 percent of cellulase and 0.03 percent of pectinase which are based on the total weight of the original magnolia leaves into the primary extraction liquid, adding the primary extraction liquid into the primary extraction residue, extracting the magnolia leaf cell water and the magnolia leaf essential oil again under the conditions of 35 ℃, pressure of-101 kPa and 80 rpm stirring, standing for 7 hours, and layering the collected liquid to obtain 36.9kg of the magnolia leaf cell water and 6.2g of the magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow, and has fresh and pleasant fragrance.
Comparative example 1:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding no solvent, extracting under stirring at 40 ℃, under the pressure of-101 kPa and 45 rpm, condensing and collecting (condensation temperature of-5 ℃) the magnolia leaf cell water and the magnolia leaf essential oil to form steam, extracting for 8 hours, separating the residues, standing and layering to obtain 28.1kg of magnolia leaf cell water and 2.7g of magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow and light in fragrance.
Comparative example 2:
putting 50kg of magnolia leaves into 150L low-temperature vacuum extraction equipment, adding 0.32% of cellulase, 0.04% of pectinase and 5kg of water based on the total weight of the original magnolia leaves, stirring for 45 minutes at 50 ℃, then extracting under stirring at 40 ℃, under the pressure of-101 kPa at 45 rpm, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam, condensing and collecting (the condensation temperature is-5 ℃) for 8 hours, separating residues, standing and layering to obtain 38.7kg of magnolia leaf cell water and 1.7g of magnolia leaf essential oil. The cell water is clear and transparent, light brown yellow, light in fragrance and has foreign flavor.
Comparative example 3:
putting 50kg of magnolia leaves into 150L extraction equipment, adding 0.32% of cellulase, 0.04% of pectinase and 5kg of water based on the total weight of the original magnolia leaves, extracting magnolia leaf cell water and magnolia leaf essential oil at 40 ℃ under normal pressure and 45 rpm stirring, extracting for 6 hours, separating residues (twice filtration, centrifugation for the first time and filtration for a 0.22 micron filter element for the second time), standing and layering to obtain 38.2kg of magnolia leaf cell water, and not obtaining magnolia leaf essential oil. The cell water is dark brown yellow, has suspended matters and has obvious peculiar smell.
Analysis of the process results of the examples and comparative examples:
the comprehensive analysis examples and comparative examples show that the extracted primary extracted cell sap is added with a certain amount of cellulase and pectinase and then poured into the primary extracted residues, the wall breaking can be accelerated under the enzymolysis effect to enable the plant cell sap to flow out, and meanwhile, the primary extracted cell sap has good permeability and can permeate into plant cells to contain more substances, so that the magnolia leaf extracting solution rich in volatile active ingredients is obtained.
Specifically, it can be seen from comparative example 1 that the weight of the extracted cell sap was low and the content of volatile active ingredients was small only by the low-temperature vacuum extraction technique. As can be seen from comparative example 2, a certain amount of enzyme and water are added for enzymolysis, and then the extraction is carried out by adopting a vacuum extraction technology, but the defects of low amount of obtained essential oil, light cell water smell and the like are caused by poor water permeability and the addition of 5kg of water. As can be seen from comparative example 3, the magnolia leaf essential oil cannot be obtained by the enzyme extraction method in the prior art, and only magnolia leaf cell water can be obtained; and the extract contains a large amount of polysaccharide, flavone and other impurities, and has low concentration and low value.
The performance test method comprises the following steps:
1. analysis of water volatile active ingredients of magnolia denudata leaf cells: the headspace gas quality was checked at 80 ℃ injection temperature. (1) Instrument information: agilent 7980A GC; MS 5975C; 50/30 μm CAR/PDMS/DVB extraction fiber head, SUPELCO USA.
(2) GC-MS conditions:
the chromatographic column is HP-INNOWAX capillary column (30m × 0.25mm × 0.25 μm); the carrier gas is He, the flow rate is 1mL/min, and the separation ratio is 5: 1; the sample injection temperature is 250 ℃; the temperature raising procedure is that the initial temperature is 40 ℃, the temperature is kept for 5min, the temperature is raised to 250 ℃ at the speed of 8 ℃/min, and the temperature is kept for 5 min.
Mass spectrum conditions: EI ionization source, energy 70 eV; the ion source temperature is 230 ℃, the quadrupole rod temperature is 150 ℃, the interface temperature is 250 ℃, and the scanning range is 30-400 m/z.
(3) Sample pretreatment: 5mL of the sample and 1g of NaCl were placed in a 20mL headspace bottle, and the cap was screwed down. After 5min of equilibrium at 80 ℃ in stirring mode, extracting for 5min at 80 ℃ with a solid phase micro-extraction needle, and then resolving for 5min at the sample inlet. The analysis results of the composition of the yulan magnolia leaf cell water of example 1 are shown in table 1.
2. The method for testing the content of flavone and polysaccharide comprises the following steps: the method for detecting the content of the total polysaccharide comprises the following steps: the phenol-sulfuric acid method is characterized in that polysaccharide is firstly hydrolyzed into monosaccharide under the action of sulfuric acid, and is quickly dehydrated to generate furfural derivatives, and then the furfural derivatives and phenol generate orange yellow compounds, so that the determination can be carried out by a colorimetric method.
Firstly, a standard curve is made, 50mg of glucose is accurately weighed and put in a 500mL volumetric flask for constant volume, 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL and 1.8mL of standard glucose solutions are respectively sucked and placed in each test tube, distilled water is respectively used for supplementing to 2.0mL, 1.0mL of 6% phenol solution and 5.0mL of concentrated sulfuric acid are sequentially added, shaking up and cooling are carried out, the mixture is placed for 20 minutes at room temperature, then the absorbance is measured at 490nm, 2.0mL of distilled water is used as a blank according to the same operation, and the operation is repeated for three times. The abscissa is the polysaccharide content and the ordinate is the absorbance value, and a standard curve is drawn.
Respectively taking magnolia leaf cell water, preparing certain concentration, measuring the absorbance value at 490nm according to the operation method, and obtaining the corresponding sugar content according to a standard curve.
The method for detecting the content of the total flavonoids comprises the following steps: the flavone mother nucleus contains basic oxygen atom, generally has phenolic hydroxyl group, can generate yellow complex with aluminum ions, and is added with sodium nitrite and sodium hydroxide to make the solution be red in alkaline solution, the solution has maximum absorption at 510nm, and the color reaction is stable within 60 min. Rutin is used as a reference substance, aluminum nitrate is used as a color developing agent for colorimetric determination of flavonoids, the absorbance and the concentration of the rutin are in a linear relationship, and the content of the total flavonoids is determined by adopting a spectrophotometry.
Accurately sucking 0, 0.4, 0.8, 1.2, 1.6, 2.0ml of rutin standard solution, putting into a 10 ml volumetric flask (marked by a written mark), and respectively adding 2.0, 1.6, 1.2, 0.8, 0.4, 0ml of 60% ethanol solution; adding 0.5ml of 5% sodium nitrite solution, shaking, and standing for 6 min; adding 0.5ml of 10% aluminum nitrate solution, and standing for 6 min; adding 4.0ml of 4% sodium hydroxide solution, adding 60% ethanol to desired volume, shaking, and standing for 15 min; (scanning for maximum absorption) the absorbance was measured at 510 nm. Taking 0.0ml as blank, taking the concentration of rutin content as abscissa, taking ordinate as corresponding absorbance under certain concentration, and making standard curve.
Sucking 1.0ml of water sample of magnolia leaf cells (adjusted according to the absorbance of the sample), putting the water sample into a 10 ml volumetric flask (marked by a written mark), and adding the water sample into 2.0ml by using 60% ethanol; adding 0.5ml of 5% sodium nitrite solution, shaking, and standing for 6 min; adding 10% aluminum nitrate, 0.5ml solution, standing for 6 min; adding 4.0ml of 4% sodium hydroxide solution, shaking, adding 60% ethanol to desired volume, and standing for 15 min; absorbance was measured at 510 nm. Calculating the content of total flavone according to the standard curve.
A is the content in the standard curve.
3. And (3) testing the water safety of the magnolia leaf cells: the HaCaT cell is a human immortal epidermal cell line, has cytotoxicity to the HaCaT cell, and can be used as reference data for safety of skin. The normal cells are in vigorous metabolism, succinate dehydrogenase in mitochondria can reduce tetrazolium salt substances into colored crystalline substances and deposit the crystalline substances around the cells, OD values can be read by an enzyme labeling instrument according to the change, and the relative growth condition of the cells can be known by comparing the OD values with a blank control group. Example 1 the results of the magnolia leaf cell water test are shown in figure 1.
The examples and comparative examples have the following performance test results:
table 1: volatile active ingredient analysis results of Yulan magnolia leaf cell water of example 1 (ingredients with low matching degree and relative content are not listed)
Table 2: table of major volatile active ingredients of yulan magnolia leaf cell water of example 1 and comparative examples 1 to 3
It can be seen from fig. 1 (data of magnolia flower leaf cell water safety test result), that magnolia flower cell water is basically non-toxic to human body skin cells.
Claims (8)
1. A method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil is characterized by comprising the following steps: the method comprises the steps of adding no solvent, primarily extracting magnolia leaves at the temperature of 30-60 ℃ and under the pressure of-60 kPa to-101 kPa, wherein the magnolia leaf cell water and the magnolia leaf essential oil form steam in the extraction process, collecting liquid through condensation, extracting for 1.5-3 hours to obtain primary extracted liquid and primary extracted magnolia leaf residue, adding 0.2-0.4% of cellulase and 0-0.1% of pectinase based on the total weight of the primary magnolia leaves into the primary extracted liquid, then adding the primary extracted liquid into the primary extracted magnolia leaf residue, extracting for 3-7 hours again under the conditions of 30-45 ℃ and under the pressure of-60 kPa to-101 kPa, and layering the collected liquid after extraction to obtain the magnolia leaf cell water and the magnolia leaf essential oil.
2. The method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil according to claim 1, wherein the primary extraction temperature is 35 ℃ to 50 ℃ and the pressure is-80 kPa to-101 kPa.
3. The method for extracting magnolia leaf cell water and magnolia leaf essential oil according to claim 1, wherein the cellulase is added in an amount of 0.25 to 0.35%, and the pectinase is added in an amount of 0.01 to 0.06%.
4. The method for extracting magnolia leaf cell water and magnolia leaf essential oil according to claim 1, wherein stirring is performed during the extraction process, and the stirring speed is 1-150 rpm.
5. The method for extracting magnolia leaf cell water and magnolia leaf essential oil according to claim 1, wherein condensation is performed during collection at a temperature of-10 ℃ to 8 ℃.
6. The method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil according to claim 1, wherein the yulan magnolia leaf is cut to have a width of 1mm to 10 mm.
7. The method for extracting yulan magnolia leaf cell water and yulan magnolia leaf essential oil according to claim 1, wherein the yulan magnolia leaf is old yulan magnolia leaf.
8. Use of the yulan magnolia leaf cell water and yulan magnolia leaf essential oil obtained by the extraction method according to any one of claims 1 to 7, characterized in that the yulan magnolia leaf cell water and yulan magnolia leaf essential oil are used in skin care products, food products and health care products.
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