阅读说明：本技术 一种天女木兰细胞水的提取方法及其应用 (Extraction method and application of Magnolia sieboldii cell water ) 是由 张文环 唐明慧 卢建仁 张兵 艾勇 何廷刚 于 2020-05-22 设计创作，主要内容包括：本发明公开了一种天女木兰细胞水的提取方法,选用天女木兰的叶、花或茎作为原料,不需添加任何溶剂,先在低温-真空(减压)条件下进行初步提取,得到初提细胞水和初提残渣,再利用初提细胞水的高渗透性,将其作为酶解溶剂,加入纤维素酶和果胶酶后,倒回初提残渣中并再次低温-真空(减压)提取得到天女木兰细胞水；工序简单,提取率高,能够保留天女木兰的有效成分。通过本发明方法得到的天女木兰细胞水,澄清透明,芬芳柔和,富含90多种挥发性活性成分；对人体安全性高,具有很好的抗氧化和抗炎效果；利用价值高,可以作为一种绿色天然的原料,应用于食品、保健品、药品及化妆品等领域。(The invention discloses a method for extracting magnolia sieboldii cell water, which comprises the steps of selecting leaves, flowers or stems of magnolia sieboldii as raw materials, performing primary extraction under the condition of low temperature-vacuum (reduced pressure) without adding any solvent to obtain primary extracted cell water and primary extracted residues, using the high permeability of the primary extracted cell water as an enzymolysis solvent, adding cellulase and pectinase, pouring the primary extracted cell water back into the primary extracted residues, and performing low temperature-vacuum (reduced pressure) extraction again to obtain magnolia sieboldii cell water; simple process, high extraction rate, and can retain effective components of Magnolia sieboldii. The magnolia sieboldii cell water obtained by the method is clear, transparent, fragrant and soft, and is rich in more than 90 volatile active ingredients; the product has high safety to human body, and has good antioxidant and anti-inflammatory effects; has high utilization value, can be used as a green and natural raw material and applied to the fields of food, health-care products, medicines, cosmetics and the like.)

1. The extraction method of the Magnolia sieboldii cell water is characterized by comprising the following steps: taking one or more of magnolia sieboldii leaves, magnolia sieboldii flowers or magnolia sieboldii stems as raw materials, adding no solvent, primarily extracting at the temperature of 30-65 ℃ and under the pressure of-60 kPa-101 kPa, forming steam by magnolia sieboldii cell water, condensing and collecting liquid, extracting for 1.5-3 hours to obtain primary extracted cell water and primary extracted residue, adding 0.2-0.5 percent of cellulase and 0-0.1 percent of pectinase which are based on the total weight of the primary raw materials into the primary extracted cell water, then pouring the primary extracted cell water back into the primary extracted residue, extracting for 4-7 hours again under the conditions of the temperature of 35-55 ℃ and the pressure of-80 kPa-101 kPa, and collecting the magnolia sieboldii cell water.

2. The extraction method of Magnolia sieboldii cell water according to claim 1, wherein the primary extraction temperature is 35-50 ℃ and the pressure is-80 kPa-101 kPa.

3. The extraction method of Magnolia sieboldii cell water according to claim 1, wherein the cellulase is added in an amount of 0.25-0.4%, and the pectinase is added in an amount of 0.02-0.07%.

4. The extraction method of Magnolia sieboldii cell water according to claim 1, wherein stirring is performed during the extraction process, and the stirring speed is 1-150 rpm.

5. The extraction method of Magnolia sieboldii cell water according to claim 1, wherein condensation is carried out during collection at a temperature of-10 ℃ to 8 ℃.

6. The extraction method of Magnolia sieboldii cell water according to claim 1, wherein Magnolia sieboldii leaves are selected as the raw material.

7. The method for extracting Magnolia sieboldii cell water according to claim 1, wherein the Magnolia sieboldii stem is a shoot with leaves or flowers connected thereto.

8. The extraction method of Magnolia sieboldii cell water according to claim 1, wherein Magnolia sieboldii leaves are cut to a width of 1mm to 10 mm; the Magnolia sieboldii stems are cut into pieces with the length of 1 cm-5 cm.

9. The method for extracting Magnolia sieboldii cellular water according to claim 1, wherein Magnolia sieboldii leaves, Magnolia sieboldii flowers or Magnolia sieboldii stems are not mildewed or rotted.

10. The application of the magnolia sieboldii cell water obtained by the extraction method of any one of claims 1 to 9 in the fields of food, health products, medicines and cosmetics.

Technical Field

The invention relates to the technical field of agricultural product treatment, in particular to an extraction method of magnolia sieboldii cell water and application thereof.

Background

Magnolia sieboldii, a small deciduous tree of the genus Magnolia of the family Magnoliaceae, has white petals, purple stamens, attractive fragrance, and a shape similar to that of Magnolia sieboldii. The flower, leaf and stem of Magnolia sieboldii contain aromatic oil. The skin care cream has strong effects on skin care, can repair weak cells, eliminate red blood streak, enhance skin tolerance, and prevent skin from stabbing pain, desquamation, red swelling, roughness and other problems, thereby improving skin allergy symptoms from root. The Magnolia sieboldii contains natural antibacterial factors, so that the skin can be far away from problems such as acne, pimple and whelk; also contains unique natural anti-inflammatory components, continuously eliminates skin inflammation, repairs damaged cells, restores skin activity and remodels the silky skin feeling. Magnolol on flowers and branches of Magnolia sieboldii can inhibit melanosome from transferring from melanocyte to keratinocyte and inhibit melanin formation, also has antioxidant effect, and is a plant for original ecology whitening. The Magnolia sieboldii also contains rare water replenishing factors and high-purity moisture-retaining components, can densely refine pores, regulate water-oil balance, enhance skin activity, and remarkably improve skin water-locking and water-storing capacity.

At the present stage, the main extraction method of the Magnolia sieboldii comprises a distillation method, an extraction method and a supercritical carbon dioxide extraction method, wherein the distillation method has long time consumption and large energy consumption, and the loss of volatile active ingredients is caused by high extraction temperature; 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 and high cost, and is not suitable for industrial production. Therefore, the development of a method for extracting magnolia sieboldii, which can retain the active ingredients of magnolia sieboldii and is efficient and convenient, is urgently needed.

Chinese patent application CN106562908A discloses a method for extracting rose cell water, which comprises subjecting rose to rotary dry distillation at 58-62 deg.C and 0.08-0.10 Mpa in a rotary evaporator, condensing and collecting a part of cell liquid permeated from rose, and collecting a part of cell liquid in a rotary evaporation bottle to be filtered; repeating the above steps for 3 times, and collecting all cell water to obtain dried flower. However, the extraction rate of the method is not high, the aromatic substances in the flowers cannot be fully extracted by simply repeating rotary steaming (the obtained dried rose flower has a small fragrance difference with the fresh rose flower, which indicates that most of the aromatic ingredients are still remained in the flowers and cannot be fully extracted), and the content of volatile active ingredients in the obtained rose cell water is not high, and the aroma is insufficient.

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 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.

Disclosure of Invention

The invention aims to provide the extraction method of the Magnolia sieboldii cell water, which has simple working procedures and high extraction rate and can keep the effective components of Magnolia sieboldii; the extracted magnolia sieboldii cell water is rich in volatile active ingredients, fragrant and soft, high in safety and high in utilization value.

The invention is realized by the following technical scheme:

a method for extracting Magnolia sieboldii cell water comprises the following steps: taking one or more of magnolia sieboldii leaves, magnolia sieboldii flowers or magnolia sieboldii stems as raw materials, adding no solvent, primarily extracting at the temperature of 30-65 ℃ and under the pressure of-60 kPa-101 kPa, forming steam by magnolia sieboldii cell water, condensing and collecting liquid, extracting for 1.5-3 hours to obtain primary extracted cell water and primary extracted residue, adding 0.2-0.5 percent of cellulase and 0-0.1 percent of pectinase which are based on the total weight of the primary raw materials into the primary extracted cell water, then pouring the primary extracted cell water back into the primary extracted residue, extracting for 4-7 hours again under the conditions of the temperature of 35-55 ℃ and the pressure of-80 kPa-101 kPa, and collecting the magnolia sieboldii cell water.

Under the condition of low-temperature vacuum, cell water and volatile components can be well exuded and evaporated, and the activity of the volatilized cell water can be retained at a lower temperature. Preferably, the temperature of the primary extraction is 35-50 ℃, and the pressure is-80 kPa-101 kPa.

According to the invention, a certain amount of enzyme is added into the primary extraction cell water, and the primary extraction residue is extracted by putting the primary extraction cell water into the container again, so that the method has the following beneficial advantages: firstly, the surface tension of primary extracted cell water is low, and the permeability is good; secondly, the pH of the primary cell extracting water 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 extraction technology. Through the synergy of the four effects, the enzymolysis speed can be controlled at a lower temperature (35-55 ℃), and the cell sap outflow speed is accelerated. In the re-extraction step, the enzymolysis speed is high during the period before the primary extraction cell water poured into the container is steamed, the enzymolysis duration is controlled by the amount of the primary extraction liquid and the technological conditions of re-extraction (at this moment, the amount of the primary extraction liquid is important, the enzymolysis time is prolonged, the enzymolysis is excessive, the enzymolysis time is shortened, the enzymolysis is insufficient), and the problems that the traditional enzymolysis method needs to add a large amount of water to dilute the extracting solution and the enzymolysis causes pungent smell are solved.

Therefore, the primary extraction time is one of the key parameters, if the time is too short, the primary extraction cell water amount is too small, the cell water is difficult to infiltrate the primary extraction residues after the enzyme is added, and the enzymolysis cannot be normally carried out; and when the cell is extracted again, the initial cell extracting water is quickly evaporated, so that the enzymolysis time is shortened; resulting in insufficient enzymatic hydrolysis. If the primary extraction time is too long, cell water flows out too much, so that the enzymolysis time is prolonged, and the risk of excessive enzymolysis is brought; and simultaneously, the subsequent extraction efficiency is reduced. The invention controls the primary cell extracting water poured into the container in the re-extraction step to be steamed within 50-70 min under the requirement condition.

Regarding the permeability of the initial extraction magnolia sieboldii cell water, experiments show that more than 50% of active ingredients such as flavone and polysaccharide can be extracted when magnolia sieboldii residue is extracted by using the magnolia sieboldii cell water, compared with the extraction method adopting pure water. The higher permeability of the Magnolia sieboldii cell water is proved.

The enzyme is macromolecular protein, can be adsorbed on the surface of the magnolia sieboldii residue cells within the required addition range, cannot volatilize under the low-temperature vacuum condition, and does not need subsequent treatment. Preferably, the addition amount of the cellulase is 0.25-0.4%, and the addition amount of the pectinase is 0.02-0.07%.

The stem of the Magnolia sieboldii is a branch connected with leaves or flowers.

The flowers, leaves and stems of the Magnolia sieboldii are rich in active ingredients. Preferably, the magnolia sieboldii leaves are selected as the raw material. Before extraction, cutting the Magnolia sieboldii leaves into 1-10 mm wide; cutting the magnolia sieboldii stem into the length of 1 cm-5 cm.

The invention selects fresh magnolia sieboldii plant raw materials which are not mildewed and rotten. Fresh Magnolia sieboldii leaves are green and full and rich in water; fresh petals of the Magnolia sieboldii are fresh, tender and succulent; the fresh magnolia sieboldii stems are rich in moisture and are not baked or dried in the sun. Generally, the fresh-keeping agent is picked up in the day of extraction, or kept fresh after being picked up by fresh-keeping means such as refrigeration and the like.

Stirring is carried out in the extraction process, and the stirring speed is 1-150 revolutions per minute. The heating is uniform, the local high temperature is prevented, and the extraction efficiency is accelerated; and in the secondary extraction step, the primary extraction cell water added with the enzyme can be uniformly mixed with the residues through stirring, so that all the primary extraction residues are fully infiltrated, and the enzymolysis is ensured.

Condensing in the collecting process, wherein the condensing temperature is-10-8 ℃.

The Magnolia sieboldii cell water extracted by the method can be applied to the fields of food, health products, medicines, cosmetics and the like.

Compared with the prior art, the invention has the following beneficial effects:

the invention extracts the Magnolia sieboldii cell water by organically combining a low-temperature vacuum (reduced pressure) technology and an enzymolysis technology, selects leaves, flowers or stems of Magnolia sieboldii as raw materials, firstly carries out primary extraction under the condition of low-temperature vacuum to obtain primary extracted cell water and primary extracted residues, then utilizes the high permeability of the primary extracted cell water as an enzymolysis solvent, adds cellulase and pectinase, then pours the primary extracted residues back, and carries out low-temperature vacuum (reduced pressure) extraction again to obtain the Magnolia sieboldii cell water. The whole extraction process does not need to add any solvent, and 100 percent of the magnolia sieboldii cell water obtained by extraction is derived from plant cells; simple process, high extraction rate, and can retain effective components of Magnolia sieboldii. Compared with a single low-temperature-vacuum (reduced pressure) extraction technology, the extraction rate of cell water is higher, and more volatile active ingredients can be extracted; meanwhile, the problem that the traditional enzymolysis method is easy to carry out enzymolysis excessively so that the extracting solution has foreign flavor is avoided.

The magnolia sieboldii cell water extracted by the method is colorless, clear and transparent, has fragrant and soft smell, is rich in more than 90 volatile active ingredients, has high safety to human bodies, and has good oxidation resistance and anti-inflammatory effects; has high utilization value, can be used as a green and natural raw material and applied to the fields of food, health-care products, medicines, cosmetics and the like.

Drawings

FIG. 1 is a bar graph of the water safety test data for Magnolia sieboldii cells 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.

Selecting fresh magnolia sieboldii leaves, magnolia sieboldii and magnolia sieboldii stems (not rotten and not mildewed) picked in the day, and cutting the magnolia sieboldii leaves into the leaves with the width of 3-5 mm; cutting the magnolia sieboldii stem into 1-2 cm long; extraction experiments were performed.

Example 1:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 40 ℃ and under the pressure of-90 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 2 hours to obtain primary extracted cell water and primary extracted residues, adding 160g of cellulase and 25g of pectinase into the primary extracted cell water, pouring into the primary extracted residues, and extracting for 6 hours again at 40 ℃ and under the pressure of-90 kPa; stirring the mixture in the whole process at 30 revolutions per minute, and condensing the mixture at-8 ℃; 41.6kg of Magnolia sieboldii cell water is collected, colorless, clear and transparent, and fragrant and soft in smell.

Example 2:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 30 ℃ and under the pressure of-90 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 2 hours to obtain primary extracted cell water and primary extracted residues, adding 160g of cellulase and 25g of pectinase into the primary extracted cell water, pouring into the primary extracted residues, and extracting for 6 hours again at 35 ℃ and under the pressure of-90 kPa; stirring the mixture in the whole process at 30 revolutions per minute, and condensing the mixture at-8 ℃; 37.7kg of Magnolia sieboldii cell water is collected, colorless, clear and transparent, and fragrant and soft in smell.

Example 3:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 40 ℃ and under the pressure of-80 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 2 hours to obtain primary extracted cell water and primary extracted residues, adding 160g of cellulase and 25g of pectinase into the primary extracted cell water, pouring into the primary extracted residues, and extracting for 6 hours again at 40 ℃ and under the pressure of-80 kPa; stirring the mixture in the whole process at 30 revolutions per minute, and condensing the mixture at-8 ℃; 35.3kg of Magnolia sieboldii cell water is collected, colorless, clear and transparent, and fragrant and soft in smell.

Example 4:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 40 ℃ and under the pressure of-90 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 2 hours to obtain primary extracted cell water and primary extracted residues, adding 125g of cellulase and 15g of pectinase into the primary extracted cell water, pouring into the primary extracted residues, and extracting for 6 hours again at 40 ℃ and under the pressure of-90 kPa; stirring the mixture in the whole process at 30 revolutions per minute, and condensing the mixture at-8 ℃; 39.2kg of Magnolia sieboldii cell water is collected, colorless, clear and transparent, and fragrant and soft in smell.

Example 5:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 55 ℃ and under the pressure of-80 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 1.5 hours to obtain primary extracted cell water and primary extracted residues, adding 160g of cellulase and 25g of pectinase into the primary extracted cell water, pouring into the primary extracted residues, and extracting for 4 hours again at 50 ℃ and under the pressure of-90 kPa; stirring at 45 r/min in the whole process, and condensing at-8 deg.C; 36.2kg of Magnolia sieboldii cell water is collected, colorless, clear and transparent, and fragrant and soft in smell.

Example 6:

weighing 50kg of Magnolia sieboldii, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 50 ℃ and under the pressure of-85 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 2.5 hours to obtain primary-extracted cell water and primary-extracted residues, adding 125g of cellulase and 30g of pectinase into the primary-extracted cell water, pouring into the primary-extracted residues, and extracting for 5 hours again at 40 ℃ and under the pressure of-95 kPa; stirring at 45 r/min in the whole process, and condensing at-8 deg.C; 43.5kg of Magnolia sieboldii cell water is collected, is clear and transparent, and has sweet and rich smell.

Example 7:

weighing 50kg of Magnolia sieboldii stems, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 65 ℃ and under the pressure of-70 kPa, forming vapor by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 3 hours to obtain primary extracted cell water and primary extracted residue, adding 185g of cellulase and 25g of pectinase into the primary extracted cell water, pouring into the primary extracted residue, and extracting for 7 hours again at 55 ℃ and under the pressure of-80 kPa; stirring at 60 r/min in the whole process, and condensing at-8 deg.C; 24.3kg of Magnolia sieboldii cell water is collected, colorless, clear and transparent, and has fresh and pleasant smell.

Comparative example 1:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, extracting at 40 deg.C and-90 kPa, condensing and collecting liquid after Magnolia sieboldii cell water forms steam, extracting for 8 hours, stirring at 30 r/min in the whole process, and condensing at-8 deg.C to obtain 31.3kg of Magnolia sieboldii cell water, which is colorless, clear and transparent, and has light fragrance.

Comparative example 2:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, mixing with 5kg of water, 160g of cellulase and 25g of pectinase, stirring for 8 hours at 40 ℃, stirring at 30 r/min, filtering by a centrifuge and then filtering by a 0.22 mu m filter membrane after extraction is finished to obtain 42.1kg of Magnolia sieboldii cell water (containing additionally added 5kg of water), wherein the liquid is yellow, the fragrance is slightly mixed with pungent odor, and the peculiar smell is heavy.

Comparative example 3:

weighing 50kg of Magnolia sieboldii, putting into 150L low-temperature extraction equipment, mixing with 5kg of water, 160g of cellulase and 25g of pectinase, stirring for 50 minutes at 40 ℃, extracting for 6 hours at 40 ℃, under-90 kPa, condensing and collecting liquid after Magnolia sieboldii cell water forms steam, stirring for 30 revolutions per minute in the whole process, and collecting to obtain 44.8kg of Magnolia sieboldii cell water (containing additional 5kg of water) at the condensation temperature of-8 ℃, wherein the Magnolia sieboldii cell water is colorless, clear and transparent, and has faint scent and mixed foreign flavor.

Comparative example 4:

weighing 50kg of Magnolia sieboldii leaves, putting into 150L low-temperature extraction equipment, adding no solvent, primarily extracting at 40 ℃ and under the pressure of-90 kPa, forming steam by Magnolia sieboldii cell water, condensing and collecting liquid, extracting for 0.5 h to obtain primary-extracted cell water and primary-extracted residues, adding 160g of cellulase and 25g of pectinase into the primary-extracted cell water, pouring into the primary-extracted residues, and extracting for 6 h again at 40 ℃ and under the pressure of-90 kPa; stirring the mixture in the whole process at 30 revolutions per minute, and condensing the mixture at-8 ℃; 36.3kg of Magnolia sieboldii cell water is collected, and the Magnolia sieboldii cell water is colorless, clear and transparent, and has light fragrance.

The test methods are as follows:

1. analysis of volatile active ingredients in aqueous solution of Magnolia sieboldii cells: the magnolia sieboldii cell water of examples and comparative examples was subjected to headspace gas detection at a sample injection temperature of 80 ℃ and analyzed for volatile substances in the cell water.

(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 test results were as follows:

table 1: example 1 analysis table of volatile active ingredients of Magnolia sieboldii cell water

As seen from the above table, the volatile components in the Magnolia sieboldii cell water mainly comprise (-) -4-terpineol (3.95%), 3-allyl-6-methoxyphenol-5-syringic acid (3.45%), ALPHA-cumyl alcohol (2.08%), beta-eucalyptol (1.89%), linalool (1.44%), eucalyptol (2.44%), most of which are essential oil components, and volatile aromatic substances. 3-allyl-6-methoxyphenol-5-syringic acid has antibacterial and antifungal effects; eucalyptol has antiasthmatic and expectorant effects.

Table 2: examples 1 to 7 and comparative examples 1 to 4 major volatile active ingredient content of Magnolia sieboldii cell water

Table 2 is shown below:

comparative analysis by examples and comparative examples: comparative example 1 adopts a single low-temperature vacuum extraction technology, extraction is insufficient, and the obtained Magnolia sieboldii cell water has less volatile active ingredients, low content and light fragrance. Comparative example 2 the problem of excessive enzymolysis occurred with the traditional pure enzymolysis method, and the obtained Magnolia sieboldii cell water contains impurities and colored substances, and has heavy foreign flavor; and water is additionally added as an enzymolysis solvent to dilute the extracting solution, so that the flavor is light and the utilization value is not high. Comparative example 3 enzymolysis is carried out firstly, and then low-temperature-vacuum (reduced pressure) technology is adopted for extraction, so that the problem of foreign flavor caused by excessive enzymolysis can be solved; and water is used as an enzymolysis solvent, and due to poor water permeability and dilution of the extracting solution by additional water, the defects of low extraction rate, light cell water flavor and the like are caused. Comparative example 4 the extraction rate was low compared to example 1, and the fragrance was weak due to the low extraction time and the low amount of cell water.

2. Magnolia sieboldii cell water antioxidant efficacy test

By DPPH free radical scavenging method, the alcoholic solution is dark purple and has strong absorption near 517nm according to the stable nitrogen free radical existing in DPPH molecules. When the free radical scavenger exists, absorption of single electrons in molecules is gradually disappeared due to pairing, the fading degree and the number of the electrons received form a quantitative relation, and therefore, the antioxidant effect of the free radical scavenging capacity determination sample can be quantitatively analyzed by determining the reduction of the absorbance at the maximum absorption wavelength of 517 nm. The magnolia sieboldii cell water of example 1 was tested for antioxidant efficacy, and the test results are shown in table 3.

Table 3: example 1 Magnolia sieboldii cell water antioxidant efficacy test data

Magnolia sieboldii cell water concentration (%)	DPPH clearance (%)
		20	12
40	17
		50	20
60	25
		80	34
100	46

As can be seen from the data in Table 3, the Magnolia sieboldii cell water has good antioxidant effect.

3. Magnolia sieboldii cell water anti-inflammatory efficacy test

Lipopolysaccharide is the main component of gram-negative bacteria cell wall, and can activate macrophage to release various inflammatory cytokines, so Lipopolysaccharide (LPS) is utilized to stimulate mouse mononuclear-macrophage to establish an in vitro inflammatory reaction model, samples are intervened, dexamethasone is used as a positive control, and enzyme-linked immunosorbent assay (ELISA) is adopted to determine the level change of inflammatory factors IL-6 and TNF-alpha, thereby discussing the in vitro anti-inflammatory action of the samples. The magnolia sieboldii cell aqueous stock solution of example 1 was formulated to different concentrations for testing, and the test results are shown in table 4.

Table 4: example 1 Magnolia sieboldii cell water anti-inflammatory efficacy test data

Magnolia sieboldii cell water concentration	IL-6(pg/mL)	TNF-α(pg/mL)
			100％	25	150
80％	29	150
			40％	32	165
15％	38	170
			10％	39	174
5％	42	176
			Blank space	25	150
LPS stimulation	50	260
			Dexamethasone	45	206

As can be seen from the data in Table 4, the Magnolia sieboldii cell water with the concentration of 5% has excellent anti-inflammatory effect compared with dexamethasone, and the IL-6 expression factor is obviously reduced after the skin is stimulated by LPS; while 80% of the leaf cell water of Magnolia sieboldii can reduce the TNF-alpha expression factor of skin stimulated by LPS to be within the normal range.

4. Test of preservation conditions of Magnolia sieboldii cell Water

The magnolia sieboldii cell water contains more active ingredients, and the preservation is very important for preventing the quality of products from being reduced due to fungus reproduction. The physicochemical values and the fungus reproduction conditions under two storage conditions of room temperature and 4 ℃ are detected for one month under the condition that a certain amount of preservative phenoxyethanol is added into the Magnolia sieboldii cell water and no preservative is added. Example 1 test results of Magnolia sieboldii cell water are shown in Table 5.

Table 5: example 1 test results of aqueous preservation conditions of Magnolia sieboldii cells

As shown in the data in Table 5, the Magnolia sieboldii cell water without preservative at room temperature is easier to grow bacteria, the Magnolia sieboldii cell water without preservative at 4 ℃ has slightly increased conductivity, but the micro-detection is within the standard range, the Magnolia sieboldii cell water with a certain amount of preservative is stored at room temperature and 4 ℃, the physicochemical data are stable, and the total number of microbial colonies reaches the standard. In conclusion, the conditions of no preservative at 4 ℃, preservative at room temperature and preservative at 4 ℃ are all suitable for the preservation of the Magnolia sieboldii cell water, and for the sake of safety, the condition of adding a certain amount of preservative at room temperature and 4 ℃ is recommended to be more stable.

5. Magnolia sieboldii cell water safety test

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. The safety of the magnolia sieboldii cell water was tested in the above method. Example 1 the results of the safety test of Magnolia sieboldii cell water are shown in figure 1.

As can be seen from the data in FIG. 1, the Magnolia sieboldii cell water is substantially non-toxic to human epidermal cells.