A yew oil with high moisturizing and soothing effects, its preparation method and application

Abstract

This invention provides a yew oil with high moisturizing and soothing effects, its preparation method, and its application. The preparation method includes the following steps: (1) subcritical extraction of yew seed powder to obtain crude yew oil; (2) molecular distillation of the obtained crude yew oil to obtain crude yew oil; (3) decolorization treatment of the crude yew oil to obtain the yew oil. The yew oil obtained by this invention has an unsaturated fatty acid content of >90%, of which the characteristic component paclitaxel (cis-5,9-octadecadienoic acid) content is >7%. In addition, it does not contain toxic and harmful components such as enaldehydes, and has good moisturizing and soothing effects.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic raw material preparation technology, specifically relating to a yew oil with high moisturizing and soothing effects, its preparation method, and its application. Background Technology

[0002] The yew, belonging to the Fabaceae family, is also known as the purple yew. It is an evergreen tree or shrub that can reach up to 30 meters in height. Due to its slow growth and difficult development, it is extremely rare and is often referred to as the "giant panda of the plant kingdom." Yew species have a wide geographical distribution, mainly in the cold temperate, temperate, and subtropical regions of the Northern Hemisphere. Currently, 11 major species have been discovered globally. my country has four species and one variety: Northeast yew, Yunnan yew, Southern yew (Taxus wallichiana), Chinese yew, and Tibetan yew (Taxus simonii). The Southern yew is endemic to my country and has the widest distribution and largest reserves among the five naturally growing yew species in the country. It is mainly distributed in the Yangtze River basin, the Nanling Mountains, and the mountains or valleys of Henan, Shaanxi (Qinling), and Gansu provinces. Jiangxi Province is also one of its important natural distribution areas.

[0003] The medicinal value of the yew tree was recorded as early as in the *Compendium of Materia Medica*, primarily used to treat cholera, typhoid fever, and for detoxification. It is also recorded in the *Modern Chinese Medicine Dictionary* and *Materia Medica Review*, where it is described as bitter, neutral, and slightly toxic, entering the heart meridian, and having the effects of reducing swelling and dissipating nodules, primarily used to treat tumors, kidney disease, and rheumatism. Modern Chinese medicine also uses it to treat diabetes, kidney disease, and gynecological diseases. Currently, over 400 compounds have been isolated from the yew plant, mainly consisting of taxanes and non-taxanes. Taxanes are taxane diterpenoids, while non-taxanes mainly include triterpenes, steroids, high-terpenes, flavonoids, phenylpropanoids, and phenols.

[0004] Currently, many scholars at home and abroad have conducted relatively detailed studies on the chemical composition of the heartwood, rhizome, and bark of the yew tree, but relatively little research has been done on the seeds of the yew tree and the yew oil obtained therefrom.

[0005] CN115844770A discloses a cosmetic containing yew extract and its preparation process. The preparation process of the cosmetic containing yew extract includes the following steps: raw material pretreatment, sun drying, pulverization, enzymatic hydrolysis, drying, grinding, mixing with water, simultaneous microwave distillation extraction, dehydration and drying, first filtration, concentration, mixing, stirring, aging, and second filtration. The prepared cosmetic containing yew extract contains yew essential oil, which contains organic substances such as paclitaxel, taxanes, and yew polysaccharides, as well as many macro- and micro-chemical elements. The cosmetic made with yew extract has the pure natural fragrance of the yew genus, a high content of active ingredients, and significant therapeutic effects on skincare. However, the drying and sun-drying processes result in low preparation efficiency and a low content of effective ingredients in the obtained extract.

[0006] CN105419931A discloses a method for extracting plant oil from plants, namely yew oil and its extraction method. The steps are as follows: using yew seeds as raw material, oil is extracted using microwave and solvent extraction methods. The oil is then removed by supercritical CO2 extraction to remove residual solvent odors and impurities. The re-extraction product under low-temperature conditions is allowed to stand, finely filtered, and refined to obtain yew oil. The use of microwave-assisted extraction, solvent extraction, and supercritical CO2 technology for re-extraction and refining not only ensures that the basic biological activity is not destroyed under low-temperature conditions but also effectively removes residual solvent odors and impurities, achieving a high standard of green quality. However, this method results in a low yield of yew oil, with a total unsaturated fatty acid content of 50-60%.

[0007] CN104388193A discloses a method for supercritical extraction of volatile oil from seeds of *Taxus wallichiana*, which mainly includes several steps such as pulverization, freeze-drying, supercritical extraction, and separation. The specific steps are: (1) pulverizing the seeds of *Taxus wallichiana* and then sieving them; (2) freeze-drying the pulverized material; (3) loading the dried raw material into an extraction vessel according to a certain filling coefficient for extraction; (4) after extraction, releasing the volatile oil from two separation vessels. This method for extracting volatile oil from *Taxus wallichiana* seeds has advantages such as high volatile oil yield, good quality, no solvent residue, simple operation, time saving, and ease of industrialization. However, this method also results in a lower yield of volatile oil from *Taxus wallichiana*, with the unsaturated fatty acids mainly being oleic acid and linoleic acid.

[0008] Therefore, developing a preparation process for yew oil that produces a light-colored oil with a high content of unsaturated fatty acids and is free of harmful alkenyl aldehydes is a key research focus in this field. Summary of the Invention

[0009] In view of the shortcomings of the prior art, the purpose of this invention is to provide a yew oil with high moisturizing and soothing effects, its preparation method and application. The yew oil obtained has a high content of unsaturated fatty acids and taxic acid, and does not contain harmful enaldehyde components.

[0010] To achieve this objective, the present invention employs the following technical solution:

[0011] In a first aspect, the present invention provides a method for preparing yew oil with high moisturizing and soothing effects, the preparation method comprising the following steps:

[0012] (1) The yew seed powder was subjected to subcritical extraction to obtain yew crude oil;

[0013] (2) The obtained crude yew oil was subjected to molecular distillation to obtain crude yew oil;

[0014] (3) The crude yew oil is decolorized to obtain the yew oil.

[0015] Preferably, the particle size of the yew seed powder is 40-100 mesh, for example, 50 mesh, 60 mesh, 70 mesh, 80 mesh, 90 mesh, etc., and preferably 60 mesh.

[0016] Preferably, the subcritical extraction uses an extractant comprising a primary extractant and a secondary extractant.

[0017] Preferably, the main extractant includes any one of ethane, propane, or butane.

[0018] Preferably, the mass percentage of the secondary extractant in the extractant is 1-10%, for example, it can be 2%, 4%, 6%, 8%, etc.

[0019] Preferably, the secondary extractant includes any one of acidic methanol, acidic ethanol, or acidic isopropanol.

[0020] Preferably, the secondary extractant contains HCl.

[0021] Preferably, the mass percentage of HCl in the secondary extractant is 3-10%, for example, it can be 4%, 5%, 6%, 7%, 8%, 9%, etc.

[0022] Preferably, the material-to-liquid ratio during subcritical extraction is 1:(10-20), for example, it can be 1:12, 1:14, 1:16, 1:18, etc. Based on 1g of yew seed powder, the volume of the extractant is 10-20mL, preferably 1:10.

[0023] Preferably, the subcritical extraction temperature is 35-50℃ (e.g., 36℃, 38℃, 40℃, 42℃, 44℃, 46℃, 48℃, etc.), preferably 45℃; the pressure is 0.1-1MPa (e.g., 0.2MPa, 0.4MPa, 0.6MPa, 0.8MPa, etc.), preferably 0.5MPa; and the extraction time is 20-80min (e.g., 25min, 30min, 35min, 40min, 45min, 50min, 55min, 60min, 65min, 70min, 75min, etc.), preferably 40min.

[0024] Preferably, the subcritical extraction is performed 1-4 times, for example, 1 time, 2 times, 3 times, or 4 times.

[0025] Preferably, the molecular distillation pressure is 40-400 Pa (e.g., 50 Pa, 100 Pa, 150 Pa, 200 Pa, 250 Pa, 300 Pa, 350 Pa, etc.), preferably 200 Pa; the temperature is 120-160℃ (e.g., 130℃, 140℃, 150℃, etc.), preferably 150℃; and the time is 45-90 min, e.g., 50 min, 60 min, 70 min, 80 min, etc.

[0026] Preferably, the material flow rate of the molecular distillation is 80-120 mL / min (e.g., 90 mL / min, 100 mL / min, 110 mL / min, etc.), preferably 90 mL / min, and the scraping speed is 30-80 r / min, e.g., 40 r / min, 50 r / min, 60 r / min, 70 r / min, etc., preferably 60 r / min.

[0027] Preferably, the decolorizing agent used in the decolorization treatment includes kaolin and / or activated carbon.

[0028] Preferably, the amount of decolorizing agent used in the decolorization treatment is 0.25-3% of the mass of crude yew oil, for example, it can be 0.5%, 1%, 1.5%, 2%, 2.5%, etc.

[0029] Preferably, the decolorization treatment is performed at a temperature of 50-90℃ (e.g., 55℃, 60℃, 70℃, 75℃, 80℃, 85℃, etc.) and for a time of 40-80 min (e.g., 45 min, 50 min, 55 min, 60 min, 65 min, 70 min, 75 min, etc.).

[0030] Preferably, the preparation method includes the following steps:

[0031] (1) The yew seed powder is subjected to subcritical extraction using any one of ethane, propane or butane, with a material-to-liquid ratio of 1:(10-20), a subcritical extraction temperature of 35-50℃, a pressure of 0.1-1MPa, an extraction time of 20-80min, and 1-4 extractions to obtain crude yew oil.

[0032] The subcritical extraction uses an extractant comprising a primary extractant and a secondary extractant. The primary extractant comprises any one of ethane, propane, or butane. The secondary extractant contains 1-10% by mass and comprises any one of acidic methanol, acidic ethanol, or acidic isopropanol. The secondary extractant contains HCl, and the HCl content in the secondary extractant is 3-10% by mass.

[0033] (2) The obtained crude yew oil was subjected to molecular distillation at 40-400 Pa and 120-160 ℃ for 45-90 min. The material flow rate of molecular distillation was 80-120 mL / min and the scraper rotation speed was 30-80 r / min to obtain crude yew oil.

[0034] (3) The crude yew oil is decolorized using white clay and / or activated carbon. The amount of decolorizing agent is 0.25-3% of the mass of the crude yew oil. The decolorization temperature is 50-90℃ and the time is 40-80min to obtain the yew oil.

[0035] Secondly, the present invention provides a yew oil with high moisturizing and soothing effects, wherein the yew oil is prepared by the preparation method described in the first aspect;

[0036] Preferably, the mass percentage of unsaturated fatty acids in the yew oil is >90%, for example, it can be 91%, 91.5%, 92%, 92.5%, 93%, 93.5%, etc.

[0037] Preferably, the paclitaxel oil contains more than 7% paclitaxel by mass, for example, 7.2%, 7.4%, 7.6%, 7.8%, 8%, etc.

[0038] The numerical range described in this invention includes not only the point values ​​listed above, but also any point values ​​within the numerical ranges not listed above. Due to space limitations and for the sake of brevity, this invention will not exhaustively list all the specific point values ​​included in the range.

[0039] Thirdly, the present invention provides the application of yew oil with high moisturizing and soothing effects as described in the second aspect in cosmetics.

[0040] Compared with the prior art, the present invention has the following beneficial effects:

[0041] 1. This invention uses a subcritical low-temperature extraction process to obtain crude yew oil. The entire process is carried out at a low temperature (≤50℃), resulting in low cost and no solvent residue. The crude yew oil obtained was subjected to molecular distillation and systematic decolorization process investigation. The final yew oil obtained is light yellow, which expands the applicability of yew oil to a certain extent.

[0042] 2. The yew oil obtained by this invention has an unsaturated fatty acid content of >90%, and the characteristic component paclitaxel (cis-5,9-octadecadienoic acid) content is >7%. In addition, it does not contain toxic and harmful components such as enaldehydes.

[0043] 3. The yew oil obtained by this invention has an extremely significant AQP-3 (aquaporin-3) expression promoting effect. At 6 μg / mL, the AQP-3 expression promotion amount is >60%, and it has good moisturizing effect.

[0044] 4. The yew oil obtained in this invention has significant inhibitory effects on TNF-α, IL-6 and NO at 60 μg / mL, and has a good soothing effect. Attached Figure Description

[0045] Figure 1 A graph showing the change in decolorization rate at different decolorization times;

[0046] Figure 2 The graph shows the change in decolorization rate at different decolorization temperatures.

[0047] Figure 3 A graph showing the change in decolorization rate for different dosages of decolorizing agent;

[0048] Figure 4 Comparative images of the appearance of yew oil in various embodiments and comparative examples;

[0049] Figure 5 The GC-MS spectrum of the crude yew oil obtained in Example 1 is shown below.

[0050] Figure 6 The GC-MS spectrum of the yew oil prepared in Example 1 is shown below.

[0051] Figure 7 Immunofluorescence assays of yew oil against AQP-3 in each embodiment and comparative example;

[0052] Figure 8 The effect of Taxus chinensis oil prepared in Examples 1-3 on the relative expression level of NO in the soothing efficacy test is shown in the figure.

[0053] Figure 9 The effect of Taxus chinensis oil prepared in Examples 1-3 on the relative expression level of TNF-α in the soothing efficacy test;

[0054] Figure 10 The effect of Taxus chinensis oil prepared in Examples 1-3 on the relative expression level of IL-6 in the soothing efficacy test. Detailed Implementation

[0055] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0056] The terms “comprising,” “including,” “having,” “containing,” or any other variations thereof, as used herein, are intended to cover non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not limited to those elements and may also include other elements not expressly listed or elements inherent to such composition, step, method, article, or apparatus.

[0057] "Optional" or "any one" means that the matter or event described thereafter may or may not occur, and the description includes both the possibility that the event will occur and the possibility that the event will not occur.

[0058] The indefinite articles “a” and “an” preceding an element or component of this invention do not impose any limitation on the quantity (i.e., number of times) of the element or component. Therefore, “an” or “a” should be interpreted as including one or at least one, and the singular form of an element or component also includes the plural form, unless the quantity clearly refers only to the singular form.

[0059] The terms "one embodiment," "some embodiments," "exemplary," "specific example," or "some examples," etc., used in this invention refer to specific features, structures, materials, or characteristics described in connection with that embodiment or example, which are included in at least one embodiment or example of the invention. In this document, the illustrative expressions of the above terms are not necessarily directed at the same embodiment or example.

[0060] In this invention, unless otherwise explicitly stated, percentages and contents are all by mass. Unless otherwise specified, the experimental methods used are conventional methods, and the materials and reagents used are commercially available.

[0061] Acidic methanol: HCl gas is dissolved in a 96% methanol aqueous solution until the HCl content in the mixture is 5%;

[0062] Acidic ethanol: HCl gas is dissolved in a 95% aqueous ethanol solution until the HCl content in the mixture is 3%;

[0063] Acidic isopropanol: HCl gas is dissolved in 99% isopropanol aqueous solution until the HCl content in the mixture is 10%.

[0064] Test Example 1

[0065] Decolorization process parameter screening

[0066] (1) Selection of decolorizing materials

[0067] Based on existing literature reports [Zhang Xu, Yuan Jiang, Wang Jiali, et al. Study on decolorization of golden sunflower oil by silica gel and activated clay [J]. Ginseng Research, 2016(6)], using the crude yew oil prepared in Example 1 as the decolorization target, the decolorization temperature was 50℃, the decolorization time was 40min, and the amount of decolorizing agent added was 2%, the effects of different composite decolorizing agents activated clay, activated carbon, and combinations of activated carbon and activated clay (1:2, 2:1, 1:1) on the decolorization effect of yew seed oil were studied;

[0068] The method for determining the decolorization rate is as follows: using distilled water as a reference, the absorbance is measured at a wavelength of 416 nm using a 1 cm cuvette. The formula for calculating the decolorization rate is: Decolorization rate (T%) = (A0 - A1) / A0;

[0069] In the formula: T is the decolorization rate; A0 is the absorbance of the oil before decolorization; A1 is the absorbance of the oil after decolorization. The decolorization results are shown in Table 1:

[0070] Table 1

[0071]

[0072] According to the data in the table, activated carbon has the highest decolorization rate at 38.02%, while activated clay has the lowest decolorization rate at 0.08%. Therefore, activated carbon is the best decolorizing agent.

[0073] (2) Effect of decolorization time on decolorization rate

[0074] Using activated carbon as the decolorizing agent, at a decolorization temperature of 70℃, and with an activated carbon dosage of 3%, the effect of different decolorization times (40 min, 50 min, 60 min, 70 min, and 80 min) on the decolorization effect of yew seed oil was investigated. The decolorization rates for different decolorization times are shown below. Figure 1 As shown in the figure, the decolorization rate of yew seed oil first increases and then decreases with the extension of decolorization time, and the decolorization effect is best when the decolorization time is 60 minutes.

[0075] (3) Effect of decolorization temperature on decolorization rate

[0076] Using activated carbon as the decolorizing agent, with a decolorization time of 60 min and an activated carbon dosage of 3%, the effect of different decolorization temperatures (50℃, 60℃, 70℃, 80℃, and 90℃) on the decolorization effect of yew seed oil was investigated. The decolorization rates at different decolorization temperatures are shown below. Figure 2 As shown in the figure, the optimal decolorization temperature is 70℃.

[0077] (4) Effect of decolorizing agent dosage on decolorization rate

[0078] Activated carbon was used as the decolorizing agent. The decolorization temperature was 70℃, and the decolorization time was 60 min. 0.25%, 0.25%, 1%, 2%, and 3% activated carbon were added to decolorize crude yew seed oil, respectively, to investigate the effect of different addition amounts on the decolorization effect. The decolorization rates of different amounts of decolorizing agent are shown below. Figure 3 As shown in the figure, the decolorization rate reached 74.42% with an added amount of 2% activated carbon, and then tended to plateau. Considering the amount of solvent used and environmental protection, the preferred amount of activated carbon added is 2%.

[0079] (5) Orthogonal experimental verification of the decolorization process:

[0080] ①The orthogonal experimental design for decolorization is shown in Table 2:

[0081] Table 2

[0082]

[0083]

[0084] ②The results of the decolorization orthogonal experiment are shown in Tables 3 and 4:

[0085] Table 3

[0086] Test A B C e Decolorization rate (%) 1 1 1 1 1 10.47 2 1 2 2 2 52.45 3 1 3 3 3 73.52 4 2 1 2 3 50.76 5 2 2 3 1 76.82 6 2 3 1 2 23.94 7 3 1 3 2 74.15 8 3 2 1 3 23.94 9 3 3 2 1 53.53 T1 45.480 45.127 19.450 46.940 / T2 50.507 51.070 52.247 50.180 / T3 50.540 50.330 74.830 49.407 / R 5.060 5.943 55.380 3.240 /

[0087] Table 4

[0088] factor Sum of squared deviations Degrees of freedom F ratio F critical value Significance Decolorization temperature 47.822 2 1.000 19.000 P>0.05 Bleaching time 58.792 2 1.229 19.000 P>0.05 Activated carbon addition amount 4601.850 2 96.227 19.000 P<0.05 error 47.82 2 / / /

[0089] Based on the data in the table and the results of the range analysis, R A <R B <R C Among the three factors, decolorization time had the least impact on the decolorization effect, followed by decolorization temperature, while the amount of decolorizing agent added had the greatest impact. Analysis of variance showed that the amount of activated carbon added during decolorization had a significant effect on the decolorization rate of yew seed oil (P < 0.05).

[0090] The optimal combination of decolorization conditions obtained from the experiment was A2B2C3 (decolorization rate: 76.82%), namely, decolorization time of 60 min, decolorization temperature of 70℃, and decolorizing agent addition of 3%. This is not significantly different from the decolorization rate result of 60 min decolorization time, 70℃ decolorization temperature, and 2% decolorizing agent addition (decolorization rate: 74.42%). Considering solvent usage and environmental protection, the final activated carbon addition amount was determined to be 2%.

[0091] Example 1

[0092] This embodiment provides a yew oil, prepared by the following method:

[0093] (1) Grinding step: Grind the Southern Yew seeds to 60 mesh and set aside;

[0094] (2) Extraction steps: Put the yew seed powder into the extraction tank and add 15 times the amount of subcritical extractant: butane + acidic methanol (w / w is 20:1). The subcritical extraction conditions are: temperature is 40℃, pressure is 0.5MPa, time is 40min, and the number of extractions is 4 times to obtain yew crude oil.

[0095] (3) Distillation step: The obtained crude yew oil was subjected to molecular distillation at 200 Pa and 150 °C for 60 min. The material flow rate of the molecular distillation was 90 mL / min and the scraper rotation speed was 60 r / min to obtain crude yew oil.

[0096] (4) Decolorization step: Add 2% activated carbon to crude yew oil, decolorize at 70℃ for 60 minutes, filter to remove the decolorizing agent, and obtain the final yew oil.

[0097] Example 2

[0098] This embodiment provides a yew oil, prepared by the following method:

[0099] (1) Grinding step: Grind the Southern Yew seeds to 40 mesh and set aside;

[0100] (2) Extraction steps: Put the yew seed powder into the extraction tank and add 10 times the amount of subcritical extractant: propane + acidic ethanol (w / w is 99:1). The subcritical extraction conditions are: temperature is 35℃, pressure is 0.3MPa, time is 40min, and the number of extractions is 3 times to obtain yew crude oil.

[0101] (3) Distillation step: The obtained crude yew oil was subjected to molecular distillation at 300 Pa and 160 °C for 50 min. The material flow rate of the molecular distillation was 80 mL / min and the scraper rotation speed was 80 r / min to obtain crude yew oil.

[0102] (4) Decolorization step: Add 2% activated carbon to crude yew oil, decolorize at 70℃ for 60 minutes, filter to remove the decolorizing agent, and obtain the final yew oil.

[0103] Example 3

[0104] This embodiment provides a yew oil, prepared by the following method:

[0105] (1) Grinding step: Grind the Southern Yew seeds to 80 mesh and set aside;

[0106] (2) Extraction steps: Put the yew seed powder into the extraction tank and add 10 times the amount of subcritical extractant: ethane + acidic isopropanol (w / w is 9:1). The subcritical extraction conditions are: temperature is 50℃, pressure is 1MPa, time is 40min, and the number of extractions is 4 times to obtain yew crude oil.

[0107] (3) Distillation step: The obtained crude yew oil was subjected to molecular distillation at 100 Pa and 120 °C for 60 min. The material flow rate of the molecular distillation was 100 mL / min and the scraper rotation speed was 40 r / min to obtain crude yew oil.

[0108] (4) Decolorization step: Add 2% activated carbon to crude yew oil, decolorize at 70℃ for 60 minutes, filter to remove the decolorizing agent, and obtain the final yew oil.

[0109] Example 4

[0110] This embodiment provides a yew oil, which differs from Example 1 only in that the extraction temperature in step (2) is adjusted to 30°C. Other raw materials, dosages and preparation methods are the same as in Example 1.

[0111] Example 5

[0112] This embodiment provides a yew oil, which differs from Example 1 only in that the extraction temperature in step (2) is adjusted to 55°C. Other raw materials, dosages and preparation methods are the same as in Example 1.

[0113] Example 6

[0114] This embodiment provides a yew oil, which differs from Example 1 only in that the subcritical extractant is butane and acidic methanol is not added. Other raw materials, dosages, and preparation methods are the same as in Example 1.

[0115] Example 7

[0116] This embodiment provides a yew oil, which differs from Example 1 only in that the subcritical extractant is butane + acidic methanol (w / w = 6:1). Other raw materials, dosages, and preparation methods are the same as in Example 1.

[0117] Example 8

[0118] This embodiment provides a yew oil, which differs from Example 1 only in that the temperature of molecular distillation in step (3) is adjusted to 100°C. Other raw materials, dosages and preparation methods are the same as in Example 1.

[0119] Example 9

[0120] This embodiment provides a yew oil, which differs from Example 1 only in that the temperature of molecular distillation in step (3) is adjusted to 180°C. Other raw materials, dosages and preparation methods are the same as in Example 1.

[0121] Comparative Example 1

[0122] This comparative example provides a yew oil, which differs from Example 1 only in that step (2) is changed to direct physical pressing to obtain crude yew oil. Other raw materials, dosages and preparation methods are the same as in Example 1.

[0123] Comparative Example 2

[0124] This comparative example provides a yew oil, which differs from Example 1 only in that step (3) is omitted, and the crude yew oil is directly decolorized in step (4). Other raw materials, dosages and preparation methods are the same as in Example 1.

[0125] Comparative Example 3

[0126] This comparative example provides a yew oil, prepared by the following method:

[0127] Referring to (Xiong Wei, Fu Jianping, Rao Yuxi, et al. Supercritical CO2 extraction of volatile oil from seeds of Taxus wallichiana and analysis of its fatty acids [J]. Food Industry, 2016(9):4), supercritical extraction was carried out under the conditions of extraction pressure of 25 MPa, extraction temperature of 46 ℃ and extraction time of 128 min to collect volatile oil from Taxus wallichiana.

[0128] Add 2% activated carbon to the volatile oil of Taxus chinensis, decolorize at 70°C for 60 minutes, filter to remove the decolorizing agent, and the final Taxus chinensis oil is obtained.

[0129] Comparative Example 4

[0130] This comparative example provides a yew oil, prepared by the following method:

[0131] Referring to (Wei Zhengbo, Xie Ying, Chen Quanbin, et al. Preliminary study on extraction and antitumor activity of Taxus chinensis seed oil [J]. Journal of Guangxi Medical University, 2009(2):4), Taxus chinensis volatile oil was obtained by Soxhlet extraction;

[0132] Add 2% activated carbon to the volatile oil of Taxus chinensis, decolorize at 70°C for 60 minutes, filter to remove the decolorizing agent, and the final Taxus chinensis oil is obtained.

[0133] Test Example 2

[0134] For example, the appearance of the yew oil obtained in Examples 1-3 and Comparative Examples 1-2 is shown in the figure below. Figure 4 As shown in the figure, the yew oil obtained by the preparation method provided by the present invention in Examples 1-3 is lighter in color and more transparent.

[0135] The absorbance of the yew oil prepared in each example and comparative example was tested at 416 nm, and the results are shown in Table 5.

[0136] Table 5

[0137]

[0138]

[0139] As shown in the table data, the absorbance of the yew oil obtained in Examples 1-3 is relatively low. As shown in Examples 1 and 4-5, the subcritical extraction temperature in the preparation process provided by this invention should not be too low. When the subcritical temperature is too low, the fluid viscosity increases and the fluidity is poor, which is not conducive to the mixing and sedimentation of the material, thus affecting the final extraction and separation effect, resulting in a higher absorbance of the final yew oil. The subcritical extraction temperature should also not be too high. When the subcritical temperature is too high, it damages the heat-sensitive components in the material, accelerates the oxidation and deterioration of some components such as unsaturated fatty acids, and darkens the color, resulting in a higher absorbance of the final yew oil.

[0140] As shown in Examples 1 and 6, the subcritical extraction method using a combination of primary and secondary extractants yields the best results. Without secondary extractants, the overall polarity of the extractant is relatively low, resulting in insufficient extraction of some relatively polar oil-soluble components, which is detrimental to subsequent decolorization and leads to higher absorbance of the final yew oil. As shown in Examples 1 and 7, excessive use of secondary extractants in the subcritical extraction method results in an overly high overall polarity of the extractant, potentially leading to the extraction of some more polar oil-soluble components. This component may be darker in color, resulting in higher absorbance of the final yew oil. As can be seen from Examples 1-3 and Examples 8-9, the pressure and temperature of molecular distillation are optimal within the range provided by this invention. If the temperature is too low, the molecular distillation will be less effective in removing color components from the crude yew oil, resulting in higher absorbance of the final yew oil. If the temperature is too high, some temperature-sensitive components will be damaged during the molecular distillation process, accelerating the oxidation and deterioration of some components, such as unsaturated fatty acids, and darkening the color, resulting in higher absorbance of the final yew oil.

[0141] As shown in Example 1 and Comparative Example 1, the yew oil prepared directly from crude yew oil obtained by physical pressing without extraction may have a darker color due to the higher content of oil-soluble impurities, making it less conducive to subsequent decolorization. Therefore, the final yew seed oil has a higher absorbance. As shown in Example 1 and Comparative Example 2, directly decolorizing the crude yew oil without molecular distillation can lead to the presence of some colored oil-soluble enaldehydes, which is also less conducive to subsequent decolorization. Therefore, the final yew seed oil has a higher absorbance. As shown in Example 1 and Comparative Examples 3-4, the yew seed oil prepared using the preparation process provided by this invention has a lighter color and lower absorbance compared to existing technologies.

[0142] Test Example 3

[0143] Chemical composition determination of yew oil

[0144] GC conditions: Aglient VF-WAXms column (30m×250μm×0.25μm); injection volume 1μL; injection port temperature 150℃; carrier gas He, split ratio 20:1; flow rate 1mL / min; initial temperature 100℃, increased to 240℃ at 10℃ / min, and held for 20min.

[0145] MS conditions: Electron impact (EI) ion source, 70 eV, ion source temperature 230 °C, mass spectrometry interface temperature 250 °C, transfer line temperature 280 °C, mass scan range (m / z) 50-500 amu.

[0146] The determination results of unsaturated fatty acids and taxic acid in the yew oils prepared in each embodiment and comparative example are shown in Table 6; for example, the GC-MS spectrum of the crude yew oil prepared in Example 1 is shown in Table 6. Figure 5 As shown, the GC-MS spectrum of the yew oil prepared in Example 1 is as follows. Figure 6 As shown in Table 7, the components of the crude yew oil and yew oil prepared in Example 1 were analyzed.

[0147] Table 6

[0148]

[0149]

[0150] Table 7

[0151]

[0152] As shown in the table, the crude yew oil obtained by the preparation method of this invention has an unsaturated fatty acid content as high as 94.08% (the total detectable fatty acid content is 100%). The yew oil obtained after molecular distillation and decolorization has an unsaturated fatty acid content of 93.01% (the total detectable fatty acid content is 98.88%). Some of these may have been converted into other fatty acids and other components that were not detected. Therefore, after molecular distillation and decolorization, the overall unsaturated fatty acid content only decreased slightly.

[0153] As shown in Examples 1 and 4-5, the subcritical extraction temperature in the preparation process provided by this invention should not be too low. When the subcritical temperature is too low, the extraction and separation effect of the material deteriorates, leading to a decrease in the content of unsaturated fatty acids and the characteristic component, paclitaxel, in the final yew oil. The subcritical extraction temperature should also not be too high. When the subcritical temperature is too high, heat-sensitive components such as unsaturated fatty acids in the crude yew oil may undergo partial oxidation and deterioration, resulting in a decrease in the content of unsaturated fatty acids and the characteristic component, paclitaxel, in the final yew oil. As shown in Examples 1 and 6, the optimal effect is achieved when the subcritical extraction uses a combination of primary and secondary extractants. When no secondary extractant is added, the overall polarity of the extractant is relatively low, resulting in insufficient extraction of some relatively polar oil-soluble components, leading to insufficient extraction of unsaturated fatty acids and the characteristic component, paclitaxel, in the final yew oil. All showed a decrease; as shown in Examples 1 and 7, when the amount of secondary extractant in the subcritical extractant is too large, the overall polarity of the extractant is relatively too high, which may lead to the extraction of some oil-soluble components with relatively higher polarity, resulting in a decrease in the content of unsaturated fatty acids and the content of the characteristic component: paclitaxel in the final yew oil; as shown in Examples 1-3 and Examples 8-9, the pressure and temperature of molecular distillation are within the range provided by the present invention, and the effect is optimal. If the temperature is too low, the molecular distillation will be less effective in removing some impurities from the crude yew oil, resulting in a decrease in the content of unsaturated fatty acids and the content of the characteristic component: paclitaxel in the final yew oil; if the temperature is too high, some temperature-sensitive components will be damaged during the molecular distillation process, accelerating the oxidation and deterioration of some components such as unsaturated fatty acids, resulting in a decrease in the content of unsaturated fatty acids and the content of the characteristic component: paclitaxel in the final yew oil.

[0154] As shown in Example 1 and Comparative Example 1, the yew oil prepared directly from crude yew oil obtained by physical pressing without extraction may contain more impurities, resulting in a decrease in unsaturated fatty acids and an inability to extract the characteristic component, paclitaxel. As shown in Example 1 and Comparative Example 2, directly decolorizing the crude yew oil without molecular distillation leads to the presence of some colored oil-soluble enaldehydes, which may not be completely removed during the decolorization process, resulting in a decrease in both the content of unsaturated fatty acids and the content of the characteristic component, paclitaxel, in the final yew oil. As shown in Example 1 and Comparative Examples 3-4, the yew oil prepared using the process provided by this invention, compared with existing technologies, can effectively extract the characteristic component, paclitaxel, and also shows a certain increase in the content of unsaturated fatty acids.

[0155] Test Example 4

[0156] Moisturizing efficacy test

[0157] AQP-3 (aquaporin-3) plays an important role in maintaining skin hydration and function. This study used immunofluorescence to determine the effect of yew oil on AQP-3 expression. HaCaT cells were seeded in 24-well plates. The experiment included a normal control group and different yew oil treatment groups (6 μg / mL). Cells were incubated in an incubator (37℃, 5% CO2) for 12 h. When the cell layer coverage reached 40%-60%, the cells were administered the oil. After administration, the cells were incubated for 24 h. Following incubation, the cells were fixed with 4% paraformaldehyde for 24 h, and then immunofluorescence was performed using a fluorescence microscope. The results are shown below. Figure 7 As shown, and using The image processing software Plus was used for analysis. The test results are shown in Table 8.

[0158] Table 8

[0159]

[0160]

[0161] The data in the table show that the yew oil prepared by this invention has a significant and excellent effect on promoting AQP-3 expression at extremely low concentrations (6 μg / mL). Examples 1 and 4-5 show that the subcritical extraction temperature in the preparation process provided by this invention should not be too low or too high; excessively high or low subcritical extraction temperatures reduce the moisturizing effect of the prepared yew oil. Examples 1 and 6 show that the optimal effect is achieved when the subcritical extraction uses a combination of primary and secondary extractants; the absence of secondary extractants affects the moisturizing effect of the yew oil. Examples 1 and 7 show that excessive use of secondary extractants in the subcritical extraction significantly reduces the moisturizing effect of the yew oil. Examples 1-3 and 8-9 show that molecular distillation temperatures within the range provided by this invention are optimal; excessively low or high temperatures will affect the moisturizing effect of the product. Examples 1-3 and Comparative Examples 3-4 show that the yew oil prepared by the process provided by this invention has a superior moisturizing effect compared to existing technologies.

[0162] Test Example 5

[0163] Soothing efficacy test

[0164] (1) Effect of the test substance on the viability of RAW264.7 cells

[0165] RAW264.7 cells with good morphology and in the logarithmic growth phase were seeded into 96-well plates and incubated for 24 hours. Five to eight concentration gradients of the test substance or a specified assay concentration were set up, diluted with complete culture medium, with three replicates for each concentration gradient, and incubated for a total of 24 hours. The original supernatant was removed, and MTT test solution was added to each well, incubated for 4 hours, and the absorbance was measured at 570 nm using a microplate reader to calculate the cell viability.

[0166] The cell viability MTT assay showed that the yew oil obtained in Examples 1-3 had a viability of ≥80% against RAW264.7 cells at 60 μg / mL. For the convenience of subsequent tests, all subsequent tests were conducted at 60 μg / mL.

[0167] (2) ELISA method was used to detect the inhibitory effects of the test substance on TNF-α, IL-6 and NO.

[0168] RAW264.7 cells in logarithmic growth phase with good morphology were seeded into 24-well plates and incubated for 24 h. A blank control group, LPS-induced stimulation group, positive control group (dexamethasone, DEX), and sample group were set up and incubated at 37℃ in a 5% CO2 incubator for 2 h. Except for the blank control group, LPS was added to each well and incubated for 24 h. The cell supernatant was analyzed according to the instructions of the ELISA kit and nitric oxide assay kit to detect the levels of NO, TNF-α, and IL-6 in the collected cell supernatant. The experimental results were analyzed using data processing software. Specific results are shown below. Figure 8-10 As shown.

[0169] As shown in the figure, the yew oil prepared in Examples 1-3 all have significant soothing effects at a low concentration (60 μg / mL).

[0170] Test Example 6

[0171] According to the "Cosmetic Safety Technical Specifications" (2015 edition), acute eye irritation tests, repeated skin irritation tests, skin allergy tests, skin phototoxicity tests, cell phototoxicity tests, and tests for hormones, microorganisms, and heavy metals were conducted; pesticide residue tests were conducted according to GB / T 39665-2020; the safety of the yew oil obtained in Examples 1-3 was jointly evaluated. The results are shown in Table 9 below, indicating that the yew oil provided by this invention is safe and non-irritating, and can be added to cosmetic formulations.

[0172] Table 9

[0173]

[0174]

[0175] The applicant declares that the present invention is illustrated by the above embodiments, but the present invention is not limited to the above process steps, that is, it does not mean that the present invention must rely on the above process steps to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent substitutions of the raw materials used in the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for preparing yew oil with high moisturizing and soothing effects, characterized in that, The preparation method includes the following steps: (1) The yew seed powder was subjected to subcritical extraction to obtain crude yew oil; The subcritical extraction uses an extractant comprising a primary extractant and a secondary extractant. The primary extractant comprises any one of ethane, propane, or butane, and the secondary extractant comprises any one of acidic methanol, acidic ethanol, or acidic isopropanol. The mass percentage of the secondary extractant in the extractant is 1-10%. The subcritical extraction temperature is 35-50℃, the pressure is 0.1-1 MPa, and the extraction time is 20-80 min. (2) The obtained crude yew oil was subjected to molecular distillation to obtain crude yew oil; The molecular distillation is carried out at a pressure of 40-400 Pa, a temperature of 120-160℃, and a time of 45-90 min. (3) The crude yew oil is decolorized to obtain the yew oil.

2. The preparation method according to claim 1, characterized in that, The particle size of the yew seed powder is 40-100 mesh.

3. The preparation method according to claim 1, characterized in that, The secondary extractant contains HCl.

4. The preparation method according to claim 3, characterized in that, The mass percentage of HCl in the secondary extractant is 3-10%.

5. The preparation method according to claim 1, characterized in that, The material-to-liquid ratio for subcritical extraction is 1 g:(10-20) mL.

6. The preparation method according to claim 1, characterized in that, The subcritical extraction is performed 1-4 times.

7. The preparation method according to claim 1, characterized in that, The material flow rate of the molecular distillation is 80-120 mL / min, and the scraping speed is 30-80 r / min.

8. The preparation method according to claim 1, characterized in that, The decolorizing agents used in the decolorization treatment include kaolin and / or activated carbon.

9. The preparation method according to claim 1, characterized in that, The amount of decolorizing agent used in the decolorization process is 0.25-3% of the mass of crude yew oil.

10. The preparation method according to claim 1, characterized in that, The decolorization process is carried out at a temperature of 50-90℃ for 40-80 minutes.

11. A yew oil with high moisturizing and soothing effects, characterized in that, The yew oil is prepared using the preparation method described in any one of claims 1-10.

12. The yew oil according to claim 11, characterized in that, The unsaturated fatty acid content in the yew oil is >90% by mass.

13. The yew oil according to claim 11, characterized in that, The yew oil contains >7% by mass of paclitaxel.

14. The use of the yew oil with high moisturizing and soothing effects as described in any one of claims 11-13 in the preparation of cosmetics.

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