Disclosure of Invention
The application aims to provide a compound essential oil, a preparation method thereof and a product for improving learning and memory abilities, so as to solve the problems.
In order to achieve the purpose, the following technical scheme is adopted in the application:
a compound essential oil, comprising: 30-35 wt% of eucalyptol and rosemary essential oil, 20-25 wt% of lemon essential oil, 10-15 wt% of peppermint essential oil, 10-15 wt% of borneol camphor essential oil, 10-15 wt% of juniper berry essential oil, 5-7 wt% of bergamot essential oil, 5-7 wt% of clove essential oil and 5-7 wt% of sandalwood essential oil.
Preferably, the compound essential oil comprises: 30-34 wt% of eucalyptol and rosemary essential oil, 20-24 wt% of lemon essential oil, 10-14 wt% of peppermint essential oil, 10-14 wt% of borneol camphor essential oil, 10-14 wt% of juniper berry essential oil, 5-6 wt% of bergamot essential oil, 5-6 wt% of clove essential oil and 5-6 wt% of sandalwood essential oil.
Preferably, the compound essential oil comprises: eucalyptol and rosemary essential oil 30 wt%, lemon essential oil 20 wt%, peppermint essential oil 15 wt%, borneol camphor essential oil 10 wt%, juniper berry essential oil 10 wt%, bergamot essential oil 5 wt%, clove essential oil 5 wt% and sandalwood essential oil 5 wt%.
A preparation method of compound essential oil comprises the following steps:
respectively pulverizing eucalyptol rosemary, lemon, peppermint, borneol camphor, juniper berries, bergamot, clove and sandalwood, and extracting to obtain eucalyptol rosemary essential oil, lemon essential oil, peppermint essential oil, borneol camphor essential oil, juniper berries essential oil, bergamot essential oil, clove essential oil and sandalwood essential oil;
mixing eucalyptol and rosemary essential oil, lemon essential oil, peppermint essential oil, borneol camphor essential oil, juniper berry essential oil, bergamot essential oil, clove essential oil and sandalwood essential oil according to a proportion to obtain the compound essential oil.
Preferably, the extracting comprises: the eucalyptol and rosemary, the peppermint, the borneol camphor, the juniper berry, the clove and the sandalwood are extracted by a steam distillation method, and the lemon and the bergamot are extracted by a squeezing method.
Preferably, the steam distillation method is used for extracting at the temperature of 100-120 ℃.
Preferably, the steam distillation method is used for extracting for 1h-3 h.
Preferably, in the steam distillation method, the steam pressure is 666.5-1333 Pa.
The application also provides a product for improving learning and memory abilities, which comprises the compound essential oil.
Compared with the prior art, the beneficial effect of this application includes:
the compound essential oil provided by the application is prepared from plant essential oil extracts containing natural active ingredients, namely eucalyptol and rosemary essential oil, lemon essential oil, peppermint essential oil, borneol camphor essential oil, juniper berry essential oil, bergamot essential oil, clove essential oil and sandalwood essential oil, wherein the d-borneol (the borneol camphor essential oil) is the core essential oil of the formula, is extracted from Chinese characteristic camphor tree varieties, and is a foundation for improving the learning and memory ability efficacy by taking bicyclic monoterpene-d-borneol as a main component. The eucalyptol rosemary, the peppermint and the clove respectively take 1, 8-cineole, the menthol and the eugenol as main components, have certain function of improving spirit, belong to different categories of oxides, alcohol and phenol, and have the fragrance and the effect of the compounds which need to be carefully matched, assist and supplement the effect of the borneol camphor, and avoid over-stimulation of cranial nerves while playing a role; the volatility of the fragrance of the borneol camphor, the eucalyptol rosemary, the peppermint and the clove is moderate, and the fragrance and the efficacy of the main body of the formula are maintained. The volatility of the aroma of the juniper berries, the lemons and the bergamots is high, the aroma is pleasant, and the juniper berries, the lemons and the bergamots are responsible for the fusion of the aroma in the formula. The sandalwood has the effects of clear thinking and mood stabilization, and meanwhile, the fragrance volatility is low, so that the sandalwood has the effect and the effect of stabilizing fragrance in the formula.
The formula is scientific in proportioning and does not add any artificial compound. Animal behavioral and histopathological experimental researches show that the essential oil prepared by the method provided by the invention can improve learning and memory behavior indexes of learning and memory disorder model mice in a Morris water maze and shuttle box, reduce the deposition condition of beta-amyloid in a hippocampal CA1 region of the brain of the model mice, and has the effect of improving learning and memory abilities. The application provides an essential oil is convenient to use, can arrange the champignon machine in or directly paint and expand fragrant use around the nasal cavity.
The preparation method of the compound essential oil provided by the application is simple in process, low in energy consumption, high in yield and excellent in quality.
The product for improving the learning and memory ability provided by the application can obtain the benefit of improving the learning and memory ability by using the compound essential oil.
Detailed Description
The terms as used herein:
"prepared from … …" is synonymous with "comprising". The terms "comprises," "comprising," "includes," "including," "has," "having," "contains," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.
The conjunction "consisting of … …" excludes any unspecified elements, steps or components. If used in a claim, the phrase is intended to claim as closed, meaning that it does not contain materials other than those described, except for the conventional impurities associated therewith. When the phrase "consisting of … …" appears in a clause of the subject matter of the claims rather than immediately after the subject matter, it defines only the elements described in the clause; other elements are not excluded from the claims as a whole.
When an amount, concentration, or other value or parameter is expressed as a range, preferred range, or as a range of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when the range "1 ~ 5" is disclosed, the ranges described should be construed to include the ranges "1 ~ 4", "1 ~ 3", "1 ~ 2 and 4 ~ 5", "1 ~ 3 and 5", and the like. When a range of values is described herein, unless otherwise stated, the range is intended to include the endpoints thereof and all integers and fractions within the range.
In these examples, the parts and percentages are by mass unless otherwise indicated.
"part by mass" means a basic unit of measure indicating a mass ratio of a plurality of components, and 1 part may represent any unit mass, for example, 1g or 2.689 g. If we say that the part by mass of the component A is a part by mass and the part by mass of the component B is B part by mass, the ratio of the part by mass of the component A to the part by mass of the component B is a: b. alternatively, the mass of the A component is aK and the mass of the B component is bK (K is an arbitrary number, and represents a multiple factor). It is unmistakable that, unlike the parts by mass, the sum of the parts by mass of all the components is not limited to 100 parts.
"and/or" is used to indicate that one or both of the illustrated conditions may occur, e.g., a and/or B includes (a and B) and (a or B).
A compound essential oil, comprising: 30-35 wt% of eucalyptol and rosemary essential oil, 20-25 wt% of lemon essential oil, 10-15 wt% of peppermint essential oil, 10-15 wt% of borneol camphor essential oil, 10-15 wt% of juniper berry essential oil, 5-7 wt% of bergamot essential oil, 5-7 wt% of clove essential oil and 5-7 wt% of sandalwood essential oil.
Preferably, the compound essential oil comprises: 30-34 wt% of eucalyptol and rosemary essential oil, 20-24 wt% of lemon essential oil, 10-14 wt% of peppermint essential oil, 10-14 wt% of borneol camphor essential oil, 10-14 wt% of juniper berry essential oil, 5-6 wt% of bergamot essential oil, 5-6 wt% of clove essential oil and 5-6 wt% of sandalwood essential oil.
Preferably, the compound essential oil comprises: eucalyptol and rosemary essential oil 30 wt%, lemon essential oil 20 wt%, peppermint essential oil 15 wt%, borneol camphor essential oil 10 wt%, juniper berry essential oil 10 wt%, bergamot essential oil 5 wt%, clove essential oil 5 wt% and sandalwood essential oil 5 wt%.
Optionally, in the compound essential oil, the content of eucalyptol and rosemary essential oil can be any value between 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt% or 30-35 wt%, the content of lemon essential oil can be any value between 20 wt%, 21 wt%, 22 wt%, 23 wt%, 24 wt%, 25 wt% or 20-25 wt%, the content of peppermint essential oil can be any value between 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt% or 10-15 wt%, the content of borneol and camphor essential oil can be any value between 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt% or 10-15 wt%, the content of juniper berry essential oil can be any value between 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt% or 10-15 wt%, the content of the bergamot essential oil can be 5 wt%, 6 wt%, 7 wt% or any value between 5 and 7 wt%, the content of the clove essential oil can be 5 wt%, 6 wt%, 7 wt% or any value between 5 and 7 wt%, and the content of the sandalwood essential oil can be 5 wt%, 6 wt%, 7 wt% or any value between 5 and 7 wt%.
A preparation method of compound essential oil comprises the following steps:
respectively pulverizing eucalyptol rosemary, lemon, peppermint, borneol camphor, juniper berries, bergamot, clove and sandalwood, and extracting to obtain eucalyptol rosemary essential oil, lemon essential oil, peppermint essential oil, borneol camphor essential oil, juniper berries essential oil, bergamot essential oil, clove essential oil and sandalwood essential oil;
mixing eucalyptol and rosemary essential oil, lemon essential oil, peppermint essential oil, borneol camphor essential oil, juniper berry essential oil, bergamot essential oil, clove essential oil and sandalwood essential oil according to a proportion to obtain the compound essential oil.
Preferably, the extracting comprises: the eucalyptol and rosemary, the peppermint, the borneol camphor, the juniper berry, the clove and the sandalwood are extracted by a steam distillation method, and the lemon and the bergamot are extracted by a squeezing method.
Preferably, the steam distillation method is used for extracting at the temperature of 100-120 ℃.
Optionally, the temperature for the steam distillation method for extraction may be any value between 100 ℃, 110 ℃, 120 ℃ or 100 ℃ and 120 ℃.
Preferably, the steam distillation method is used for extracting for 1h-3 h.
Optionally, the time for the steam distillation method to extract can be any value between 1h, 2h, 3h or 1h and 3 h.
Preferably, in the steam distillation method, the steam pressure is 666.5-1333 Pa.
Alternatively, in the steam distillation method, the steam pressure may be any value between 666.5Pa, 700Pa, 800Pa, 900Pa, 1000Pa, 1100Pa, 1200Pa, 1300Pa, 1333Pa, or 666.5 to 1333 Pa.
The application also provides a product for improving learning and memory abilities, which comprises the compound essential oil.
Embodiments of the present application will be described in detail below with reference to specific examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present application and should not be construed as limiting the scope of the present application. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Example 1
This embodiment provides a compound essential oil, including: eucalyptol and rosemary essential oil 30 wt%, lemon essential oil 20 wt%, peppermint essential oil 15 wt%, borneol camphor essential oil 10 wt%, juniper berry essential oil 10 wt%, bergamot essential oil 5 wt%, clove essential oil 5 wt% and sandalwood essential oil 5 wt%.
The embodiment also provides a method for preparing the compound essential oil, which comprises the following steps:
(1) respectively pulverizing eucalyptol rosemary, lemon, peppermint, borneol camphor, juniper berries, bergamot, clove and sandalwood, extracting the eucalyptol rosemary, the peppermint, the borneol camphor, the juniper berries, the clove and the sandalwood by a steam distillation method, and extracting the lemon and the bergamot by a squeezing method to obtain essential oil of the eucalyptol rosemary, the lemon, the peppermint, the borneol camphor, the juniper berries, the bergamot, the clove and the sandalwood; the extraction temperature of the steam distillation method is 100 ℃, and the extraction time is 3 h.
(2) Mixing the eucalyptol rosemary, the lemon, the peppermint, the borneol camphor, the juniper berry, the bergamot, the clove and the sandalwood essential oil obtained in the step (1) according to the proportion to obtain the compound essential oil.
Example 2
This embodiment provides a compound essential oil, including: eucalyptol and rosemary essential oil 30 wt%, lemon essential oil 25 wt%, peppermint essential oil 10 wt%, borneol camphor essential oil 10 wt%, juniper berry essential oil 10 wt%, bergamot essential oil 5 wt%, clove essential oil 5 wt% and sandalwood essential oil 5 wt%.
The preparation method is the same as the example 1 except that the extraction temperature of the steam distillation method is 110 ℃ and the extraction time is 2 h.
Example 3
This embodiment provides a compound essential oil, including: 31 wt% of eucalyptol and rosemary essential oil, 21 wt% of lemon essential oil, 10 wt% of peppermint essential oil, 11 wt% of borneol camphor essential oil, 10 wt% of juniper berry essential oil, 6 wt% of bergamot essential oil, 6 wt% of clove essential oil and 5 wt% of sandalwood essential oil.
The preparation method is the same as the example 1 except that the extraction temperature of the steam distillation method is 120 ℃ and the extraction time is 1 h.
In order to better evaluate the efficacy of the compound essential oil provided by the application, the following experiments are carried out:
first, an experiment for improving learning and memory ability
Animal behavior experiments after the compound essential oil obtained in example 1 is applied to mice prove that the compound essential oil provided by the application has the efficacy of improving the learning and memory abilities.
Meanwhile, a corresponding control group is arranged for carrying out a control experiment.
1. Laboratory animal
SPF-class male 6-month-old APP/PS1 double-transgenic learning and memory disorder model mice 24, SPF-class male 8 normal mice of the same age and genetic background purchased from Kyoho Kavens laboratory animals under license number SCXK (Su) 2016-0010. Feeding conditions are as follows: in the light and shade, the temperature is about 22 ℃ and the people can freely drink water and eat the food.
2. Experiment grouping
24 model mice were divided into model group, compound essential oil group and positive group at random, and 8 normal mice were used as normal group. The compound essential oil smells compound essential oil, and the positive group smells rosemary essential oil (proved by experiments of a plurality of scholars, the compound essential oil has the efficacy of improving learning and memory). The sniffing method comprises placing fragrance lamp containing 3% volume fraction essential oil water solution in a closed box, placing mouse after essential oil volatilizes and diffuses, covering the box, and sniffing for 1 hr for 2 times per day for 21 days. Model and normal groups were untreated.
3. Experimental methods
3.1 Morris Water maze experiment
And the 12 th to 16 th days after the essential oil is sniffed is a positioning navigation experiment, the water maze comprises quadrants I, II, III and IV, a platform is placed in a certain quadrant, and the platform is slightly lower than the water surface. The mice were placed in a pool from one quadrant and the time from entry to successful landing was recorded as the escape latency. If the station is not successfully staged for more than 120 seconds, the stage is marked as 120 seconds. Experiments were performed 2 times per day at the start of the different quadrants. And day 17 is a space exploration experiment, the platform is removed, water enters from the starting point of a certain quadrant, and the times and the residence time of the mouse passing through the original platform in the quadrant where the original platform is located within 120 seconds are recorded.
3.2 shuttle experiment
The 20 th day after the essential oil is smelled and inhaled is a learning and training stage, a mouse is placed into one chamber and stimulated by light and buzzer, if the mouse successfully escapes to the other chamber within 10 seconds, the mouse is marked as primary active avoidance, and the used time is an active avoidance latency period; if the mouse is still in the home chamber after 10 seconds, the electrical stimulus is given to force the mouse to escape to the other chamber. Each mouse was cycled 20 times. After 24 hours (day 21) is the memory test phase, the procedure is the same as before.
4. Results of the experiment
4.1 Morris Water maze test results
Positioning navigation experiment results:
as shown in figure 1 (note: compared to the normal group,*P<0.05,**P<0.01; in comparison with the set of models,#P<0.05,##P<0.01) and starting from the 3 rd day of the positioning voyage experiment, the escape latencies of the model group are obviously higher than those of the normal group, and the escape latencies of the compound essential oil group are obviously lower than those of the model group, which shows that the compound essential oil improves the learning and memory disorder of the model mouse.
The experimental result of space exploration is as follows:
as shown in FIG. 2In the aspect of times of crossing the original platform, the model group is less than that of the normal group, and the compound essential oil is more than that of the model group, so that the expected result is met, but no significant difference exists. As shown in fig. 3, the model group was less than the normal group and the compound essential oil group was more than the model group in terms of residence time in the target quadrant, which was consistent with the expected results but without significant differences. Note: in comparison with the normal group,*P<0.05,**P<0.01; in comparison with the set of models,#P<0.05,##P<0.01。
4.2 shuttle test results
As shown in figure 4, in terms of the number of active avoidance, the number of active avoidance of the model mouse is greatly increased by the compound essential oil in the two stages of learning training and memory testing. As shown in fig. 5, in the aspect of active avoidance latency, in the learning training stage, the compound essential oil greatly reduces the active avoidance latency of the model mouse (P <0.01), and there is no significant difference among groups in the memory testing stage.
4.3, summary
The compound essential oil improves the learning and memory behaviors of the learning and memory disorder mice by combining 3 indexes of the escape latency, the original platform crossing times and the target quadrant residence time of the Morris water maze experiment and 2 indexes of the active avoidance times and the active avoidance latency of the shuttle experiment.
Second, improving learning and memory disorder experiment
Histopathology experiments were performed after the compound essential oil obtained in example 1 was applied to mice to prove that the compound essential oil provided in the present application has an effect of improving learning and memory disorders.
1. Laboratory animal
SPF-class male 6-month-old APP/PS1 double-transgenic learning and memory disorder model mice 24, SPF-class male 8 normal mice of the same age and genetic background purchased from Kyoho Kavens laboratory animals under license number SCXK (Su) 2016-0010. Feeding conditions are as follows: in the light and shade, the temperature is about 22 ℃ and the people can freely drink water and eat the food.
2. Experiment grouping
24 model mice were divided into model group, compound essential oil group and positive group at random, and 8 normal mice were used as normal group. The compound essential oil smells compound essential oil, and the positive group smells rosemary essential oil (proved by experiments of a plurality of scholars, the compound essential oil has the efficacy of improving learning and memory). The sniffing method comprises placing fragrance lamp containing 3% volume fraction essential oil water solution in a closed box, placing mouse after essential oil volatilizes and diffuses, covering the box, and sniffing for 1 hr for 2 times per day for 21 days. Model and normal groups were untreated.
3. Experimental methods
After the sniffing is completed. The method comprises the following steps of taking a mouse brain tissue, carrying out conventional immunohistochemical experiment steps of embedding the tissue, slicing, fishing the tissue, dewaxing, antigen retrieval, serum sealing, adding a primary antibody, adding a secondary antibody, adding SABC, adding a color developing agent, counterstaining, dehydrating and sealing, and observing the beta-amyloid deposition condition of the mouse brain hippocampus CA1 area under a microscope.
4. Results of the experiment
As shown in fig. 6, a. is the normal group, B is the model group, C is the compound essential oil group, D is the positive group; arrows indicate the surrounding beta-amyloid. No obvious beta-amyloid is observed around neuron cells in the hippocampal CA1 region of the brain of a normal group of mice, the model group has brown obvious beta-amyloid, and the compound essential oil group and the positive group have faint yellow beta-amyloid. The staining intensity shows that the compound essential oil beta-amyloid protein deposition is less than that of the model group, which indicates that the compound essential oil improves the brain histopathological conditions related to the learning and memory of mice.
Third, brain wave experiment
The brain electricity is the synthesis of postsynaptic potential of brain neurons, reflecting the electrical parameters of brain electrical activity, brain functional state and brain neural activity. Common brain waves are divided into fast waves (beta and gamma waves increase in proportion when waking, and memory and thinking are active), medium waves (alpha waves increase in proportion when eyes are quiet and closed), and slow waves (delta and theta waves increase in proportion when sleeping). 40 volunteers are recruited, and brain wave change conditions are respectively tested and analyzed before smelling the fragrance and after smelling the compound fragrance for 15 minutes. The results are shown in table 1 below:
TABLE 1 electroencephalogram changes
Test group
|
Alpha wave (%)
|
Beta wave (%)
|
Delta wave (%)
|
Theta wave (%)
|
Before smelling fragrance
|
44.15±4.24
|
23.37±3.17
|
15.66±2.18
|
16.82±1.84
|
After smelling the fragrance
|
43.63±5.71
|
25.16±3.22**
|
13.98±2.43*
|
16.23±2.36 |
As can be seen from table 2 above, the proportion of the beta wave increases before and after smelling the fragrance of the compound essential oil, and there is a very significant difference (p <0.01), which indicates that the brain wave representing the excitation reaction and the thinking activity is excited; the decrease in the delta wave ratio, indicative of a decrease in the fatigue brain wave ratio, showed significant differences (p < 0.05). The smell of the compound essential oil fragrance can activate thinking, improve memory and relieve brain fatigue.
Fourth, sensory evaluation test
100 volunteers of different sexes and ages are recruited, and the fragrance of 6 single essential oils and compound essential oils with refreshing effect, namely eucalyptol and rosemary essential oil, lemon essential oil, peppermint essential oil, borneol camphor essential oil, juniper berry essential oil and clove essential oil, is evaluated through a fragrance preference evaluation table (the score is 0-9). The single essential oils of eucalyptol, rosemary, peppermint, juniper berry, clove and lemon are common essential oils with refreshing effect, but have disadvantages. Eucalyptol rosemary has low fragrance liking degree, and is not suitable for people with high blood pressure when being used alone. The peppermint and clove have high fragrance ratio, strong irritation and obvious preference degree dipolar differentiation. The natural sense of the aroma of the juniper berry essential oil is weak, and the preference degree of the aroma is general. The borneol camphor is a unique camphor tree variety in China, is a new variety in essential oil resources, and is also a characteristic essential oil in the formula.
The evaluation results are shown in table 2 below:
TABLE 2 sensory evaluation results
As can be seen from table 2 above, the compound essential oil is superior to other single essential oils in evaluation of natural aroma, refreshing aroma, and preference of aroma, which indicates that the compound formulation achieves a good synergistic effect.
Comparative example 1
The comparative example provides a compound essential oil comprising: eucalyptol rosemary essential oil 31 wt%, lemon essential oil 25 wt%, peppermint essential oil 15 wt%, juniper berry essential oil 12 wt%, bergamot essential oil 6 wt%, clove essential oil 6 wt% and sandalwood essential oil 5 wt%.
The lack of the cinnamomum camphora essential oil can directly affect the stability of the proportion data of the beta waves in the brain wave experiment of the volunteers, and the change does not reflect the difference, as shown in table 3, and the proportion of the beta waves is the most important index for representing the active thinking and reflecting the learning and memory states.
TABLE 3 electroencephalogram change
The compound essential oil provided by the application has the effect of improving the learning and memory ability. The compound essential oil is extracted from various natural plants, has no any artificial compound, and has high safety. The essential oil has scientific formula, and the effects are verified through animal behaviours and histopathology experiments, so that the learning and memory ability can be effectively improved. In addition, the essential oil has the characteristics of high stability and convenience in use, and has popularization and application values.
Finally, it should be noted that: the above embodiments are only used for illustrating the technical solutions of the present application, and not for limiting the same; although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present application.
Moreover, those skilled in the art will appreciate that while some embodiments herein include some features included in other embodiments, rather than other features, combinations of features of different embodiments are meant to be within the scope of the application and form different embodiments. For example, in the claims above, any of the claimed embodiments may be used in any combination. The information disclosed in this background section is only for enhancement of understanding of the general background of the application and should not be taken as an acknowledgement or any form of suggestion that this information forms the prior art already known to a person skilled in the art.