CN105687368B - Application of jujube kernel volatile oil in mouse hair growth - Google Patents
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Abstract
The invention discloses an application of jujube kernel volatile oil in the hair growth of mice, the jujube kernel volatile oil is used as an active ingredient for promoting the hair growth of the mice, and the administration by gastric gavage and the smearing of the part to be grown are carried out simultaneously. The invention can promote the rapid growth of the hair of the mouse.
Description
Technical Field
The invention belongs to the technical field of extraction of active ingredients of natural plants, and particularly relates to application of jujube kernel volatile oil in mouse hair growth.
Background
The semen Ziziphi Spinosae (Zizyphus jujube Mill) is dry mature kernel of Zizyphus in Zizyphus of Rhamnaceae, and mainly contains compounds such as jujube benevolence saponins, triterpenes, fatty oil, protein, sterols, phospholipids, etc. Modern pharmacological studies show that the jujube kernel has the effects of tonifying heart, tranquilizing, calming nerves, arresting sweating, tonifying liver, promoting the production of body fluid and the like, is used for treating symptoms such as dysphoria, insomnia, fright dreaminess, body deficiency, hyperhidrosis, body fluid deficiency, thirst and the like, and is a common sedative and hypnotic medicine. It is listed as the superior in Shen nong Ben Cao Jing (Shen nong's herbal Jing), and as the root system in Ben Cao gang mu (compendium of materia Medica). The medical books are originally seen in treatise on febrile diseases and famous in jujube kernel decoction, and have been in medicinal history for over 1700 years. The recent generation is recorded in the pharmacopoeia of the people's republic of China, a calendar edition. The volatile oil is one of the main active ingredients of the jujube kernel, and in recent years, the jujube kernel oil has been researched to have the same sedative-hypnotic effect as the jujube kernel gleditsia sinensis components, and is gradually an important research content of the jujube kernel.
At present, the extraction of jujube kernel volatile oil is roughly realized by the following methods: steam distillation, organic solvent extraction and supercritical fluid extraction. The former two methods have the problems of low yield, complicated process, unstable operation conditions and the like, and the supercritical CO2The fluid extraction method is a new extraction and separation technology at present, is particularly suitable for extracting thermosensitive substances, andand the method has the advantages of no heavy metal pollution, no solvent residue, convenient transportation, stable extraction conditions and the like, and the yield of the target product can be effectively improved by adding a proper amount of entrainer in the extraction process, but the problem of low extraction oil yield still exists.
At present, the cultivation area of the Xinjiang jujube trees reaches 700 ten thousand mu, rich red jujube resources provide unique and advantageous conditions for the development and research of jujube kernels, and the processing of red jujubes has become an important link for the large-scale production of the Xinjiang forest fruit planting industry and the income increase of farmers; however, the development of new application of the jujube kernel volatile oil is less.
Disclosure of Invention
The invention aims to provide application of jujube kernel volatile oil capable of promoting the rapid growth of mouse hair in the growth of mouse hair.
The invention is realized by the following technical scheme:
the application of jujube kernel volatile oil in the growth of mouse hair is characterized by that the jujube kernel volatile oil is used as active component for promoting the growth of mouse hair, and the administration by gavage and smearing on the hair-growing portion are simultaneously implemented.
The application of the jujube kernel volatile oil in the hair growth of mice is characterized in that the dosage of the jujube kernel volatile oil for stomach irrigation administration of the mice is 1-3g/Kg, the parts to be grown of the mice are coated with an external hair growth preparation taking the jujube kernel volatile oil as an active ingredient, the external hair growth preparation consists of glycerin, propylene glycol and the jujube kernel volatile oil, the volume ratio of the glycerin to the propylene glycol is 3:4-8, and the content of the jujube kernel volatile oil in the external hair growth preparation is 50-100 mg/mL.
The application of the jujube kernel volatile oil in the hair growth of mice is characterized in that the jujube kernel volatile oil is prepared by the following steps:
(1) pre-treating the jujube kernel raw material: oven drying semen Ziziphi Spinosae, pulverizing, and sieving; the drying temperature of the jujube kernels is 45-65 ℃, the drying time is 15-25h, and the granularity of the crushed and sieved powder is 50-70 meshes;
(2) charging: adding the pretreated semen Ziziphi Spinosae into supercritical CO2In an extraction kettle of a fluid extraction device;
(3) setting the temperature and pressure of the extraction kettle, the separation kettle I and the separation kettle II; the extraction pressure of the extraction kettle is 20-35MPa, the extraction temperature is 35-50 ℃, the static extraction time is 0.5-1h, the dynamic primary extraction time is 1-2h, and the dynamic secondary extraction time is 2-3 h; the separation pressure of the separation kettle I is 8.0-9.0MPa, the separation temperature is 30-40 ℃, and the separation time is 1-3 h; the separation pressure of the separation kettle II is 4.9-5.8MPa, the separation temperature is 30-40 ℃, and the separation time is 0.5-2 h;
(4) static extraction: introducing a certain amount of CO2Raising the pressure in the extraction kettle to a preset pressure, and firstly, taking the solvent A as an entrainer to perform static extraction;
(5) dynamic primary extraction: CO regulation2The solvent A is taken as an entrainer to carry out dynamic primary extraction;
(6) dynamic secondary extraction: CO regulation2Finally adding a solvent B as an entrainer to perform dynamic secondary extraction;
(7) mixing the jujube kernel volatile oil obtained in the separation kettle I and the separation kettle II;
(8) and (5) separating and purifying the jujube kernel volatile oil obtained in the step (7) by column chromatography, and removing a mobile phase to obtain a pure jujube kernel volatile oil product.
The application of the jujube kernel volatile oil in the hair growth of mice comprises the following steps (4) and (5): the solvent A is one of n-hexane, ethyl acetate and chloroform.
The application of the jujube kernel volatile oil in the mouse hair growth comprises the following steps of (6): the solvent B is n-butyl alcohol, tert-butyl methyl ether or a mixture of n-butyl alcohol and tert-butyl methyl ether.
The application of the jujube kernel volatile oil in the hair growth of mice comprises the following steps (5) and (6): CO 22The flow rates of the water-based fuel are all 1.0-1.5L/min.
The application of the jujube kernel volatile oil in the growth of mouse hair is characterized in that the dosage of the solvent A is 20-30 wt% of the jujube kernel raw material, and the dosage of the solvent B is 50-100 wt% of the jujube kernel raw material.
The application of the jujube kernel volatile oil in the mouse hair growth is carried out in the step (8): a) a chromatographic column: ACQUITYUPLC BEH C18 column, 2.1mm × 50mm,1.7 μm; b) the mobile phase consists of a phase A and a phase B: the phase A is as follows: methanol, phase B is: 1mmol/L ammonium formate aqueous solution; c) gradient elution conditions: the volume ratio of the phase A to the phase B is 10:1 at the beginning, the volume ratio of the phase A to the phase B is 8:3 at 1 minute, the volume ratio of the phase A to the phase B is 7:4 at 3 minutes, the volume ratio of the phase A to the phase B is 6:5 at 6 minutes, the volume ratio of the phase A to the phase B is 5:6 at 9 minutes, the volume ratio of the phase A to the phase B is 4:7 at 12 minutes, the volume ratio of the phase A to the phase B is 3:8 at 15 minutes, the volume ratio of the phase A to the phase B is 1:10 at 18 minutes, and the collection starting time is as follows: 1min, end of collection time: 18 min; d) column temperature: 35 ℃; e) flow rate: 0.6 mL/min.
The application of the jujube kernel volatile oil in the hair growth of mice also comprises the steps of sequentially carrying out alkali soaking, acid soaking, buffer solution soaking and air drying on the crushed and sieved jujube kernel raw material in the step (1); the alkali solution used for alkali soaking is a sodium carbonate solution with the mass concentration of 5-12 wt%, the dosage of the alkali solution per 100g of jujube kernel powder is 300-400mL, the alkali soaking time is 5-10h, and the filtering is carried out after the alkali soaking; the acid solution used for acid soaking is hydrochloric acid with the mass concentration of 10-15 wt%, the dosage of the alkali solution of each 100g of jujube kernel powder is 200-300mL, the acid soaking time is 0.5-2h, and the filtering is carried out after the acid soaking; the buffer solution used for soaking is phosphoric acid buffer solution with pH 7.2, and the soaking time of the buffer solution is 5-10 h.
The invention has the beneficial effects that: the invention can promote the rapid growth of the hair of the mice, increase the medicinal value and the practical value of the jujube kernel volatile oil, improve the utilization rate of the jujube kernel and increase the economic benefit of the jujube kernel volatile oil.
Detailed Description
Preparation example of jujube kernel volatile oil
Example 1:
the application of jujube kernel volatile oil in the hair growth of mice comprises the following steps:
(1) pre-treating the jujube kernel raw material: oven drying semen Ziziphi Spinosae at 45 deg.C for 15 hr, pulverizing, and sieving to obtain powder with particle size of 50-70 mesh;
(2) charging: adding the pretreated semen Ziziphi Spinosae material and solvent A into supercritical CO2In an extraction kettle of a fluid extraction device; in the embodiment, the solvent A is n-hexane; the dosage of the solvent A is 20 wt% of the mass of the jujube kernel raw material;
(3) setting the temperature and pressure of the extraction kettle, the separation kettle I and the separation kettle II;
the extraction pressure of the extraction kettle is 20MPa, and the extraction temperature is 35 ℃; the separation pressure of the separation kettle I is 8.0MPa, and the separation temperature is 30 ℃; the separation pressure of the separation kettle II is 4.9MPa, and the separation temperature is 30 ℃;
(4) static extraction: introducing a certain amount of CO2Raising the pressure in the extraction kettle to the set pressure in the step (3), and firstly, taking the solvent A as an entrainer to perform static extraction for 0.5 h;
(5) dynamic primary extraction: CO regulation2The flow rate of the solvent A is 1.0L/min, and then the solvent A is taken as an entrainer to carry out dynamic primary extraction for 1 h;
(6) dynamic secondary extraction: CO regulation2The flow rate of the extraction kettle is 1.0L/min, and finally, a solvent B is added into the extraction kettle by an entrainer pump to be used as an entrainer for dynamic secondary extraction, wherein the dynamic secondary extraction time is 2 hours; the solvent B is n-butyl alcohol, and the dosage of the solvent B is 50 wt% of the mass of the jujube kernel raw material;
(7) separating in a separation kettle I for 1h and a separation kettle II for 0.5h, and mixing the jujube kernel volatile oil obtained in the separation kettle I and the separation kettle II;
(8) and (3) separating and purifying the jujube kernel volatile oil obtained in the step (7) by column chromatography, wherein a) the column chromatography: an ACQUITY UPLC BEHC18 column, 2.1mm × 50mm,1.7 μm; b) the mobile phase consists of a phase A and a phase B: the phase A is as follows: methanol, phase B is: 1mmol/L ammonium formate aqueous solution; c) gradient elution conditions: the volume ratio of the phase A to the phase B is 10:1 at the beginning, the volume ratio of the phase A to the phase B is 8:3 at 1 minute, the volume ratio of the phase A to the phase B is 7:4 at 3 minutes, the volume ratio of the phase A to the phase B is 6:5 at 6 minutes, the volume ratio of the phase A to the phase B is 5:6 at 9 minutes, the volume ratio of the phase A to the phase B is 4:7 at 12 minutes, the volume ratio of the phase A to the phase B is 3:8 at 15 minutes, the volume ratio of the phase A to the phase B is 1:10 at 18 minutes, and the collection starting time is as follows: 1min, end of collection time: 18 min; d) column temperature: 35 ℃; e) flow rate: 0.6 mL/min; removing mobile phase to obtain pure Zizyphi Spinosae semen volatile oil.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 21.15%.
Example 2:
the application of jujube kernel volatile oil in the hair growth of mice comprises the following steps:
(1) pre-treating the jujube kernel raw material: oven drying semen Ziziphi Spinosae at 65 deg.C for 25 hr, pulverizing, and sieving to obtain powder with particle size of 50-70 mesh;
(2) charging: adding the pretreated semen Ziziphi Spinosae material and solvent A into supercritical CO2In an extraction kettle of a fluid extraction device; in this example, solvent a is ethyl acetate; the dosage of the solvent A is 30 wt% of the mass of the jujube kernel raw material;
(3) setting the temperature and pressure of the extraction kettle, the separation kettle I and the separation kettle II;
the extraction pressure of the extraction kettle is 35MPa, and the extraction temperature is 50 ℃; the separation pressure of the separation kettle I is 9.0MPa, and the separation temperature is 40 ℃; the separation pressure of the separation kettle II is 5.8MPa, and the separation temperature is 40 ℃;
(4) static extraction: introducing a certain amount of CO2Raising the pressure in the extraction kettle to the set pressure in the step (3), and firstly, taking the solvent A as an entrainer to perform static extraction for 1 h;
(5) dynamic primary extraction: CO regulation2The flow rate of the solvent A is 1.5L/min, and then the solvent A is taken as an entrainer to carry out dynamic primary extraction for 2 hours;
(6) dynamic secondary extraction: CO regulation2The flow rate of the extraction kettle is 1.5L/min, and finally, a solvent B is added into the extraction kettle by an entrainer pump to be used as an entrainer for dynamic secondary extraction, wherein the dynamic secondary extraction time is 3 hours; the solvent B is tert-butyl methyl ether, and the using amount of the solvent B is 100 wt% of the mass of the jujube kernel raw material;
(7) separating in a separation kettle I for 3h and a separation kettle II for 2h, and mixing the jujube kernel volatile oil obtained in the separation kettle I and the separation kettle II;
(8) and (3) separating and purifying the jujube kernel volatile oil obtained in the step (7) by column chromatography, wherein a) the column chromatography: an ACQUITY UPLC BEHC18 column, 2.1mm × 50mm,1.7 μm; b) the mobile phase consists of a phase A and a phase B: the phase A is as follows: methanol, phase B is: 1mmol/L ammonium formate aqueous solution; c) gradient elution conditions: the volume ratio of the phase A to the phase B is 10:1 at the beginning, the volume ratio of the phase A to the phase B is 8:3 at 1 minute, the volume ratio of the phase A to the phase B is 7:4 at 3 minutes, the volume ratio of the phase A to the phase B is 6:5 at 6 minutes, the volume ratio of the phase A to the phase B is 5:6 at 9 minutes, the volume ratio of the phase A to the phase B is 4:7 at 12 minutes, the volume ratio of the phase A to the phase B is 3:8 at 15 minutes, the volume ratio of the phase A to the phase B is 1:10 at 18 minutes, and the collection starting time is as follows: 1min, end of collection time: 18 min; d) column temperature: 35 ℃; e) flow rate: 0.6 mL/min; removing mobile phase to obtain pure Zizyphi Spinosae semen volatile oil.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 23.11%.
Example 3:
the application of jujube kernel volatile oil in the hair growth of mice comprises the following steps:
(1) pre-treating the jujube kernel raw material: oven drying the semen Ziziphi Spinosae at 50 deg.C for 17 hr, pulverizing, and sieving to obtain powder with particle size of 50-70 mesh;
(2) charging: adding the pretreated semen Ziziphi Spinosae material and solvent A into supercritical CO2In an extraction kettle of a fluid extraction device; in the embodiment, the solvent A is one of trichloromethane; the dosage of the solvent A is 22 wt% of the mass of the jujube kernel raw material;
(3) setting the temperature and pressure of the extraction kettle, the separation kettle I and the separation kettle II;
the extraction pressure of the extraction kettle is 30MPa, and the extraction temperature is 40 ℃; the separation pressure of the separation kettle I is 8.5MPa, and the separation temperature is 35 ℃; the separation pressure of the separation kettle II is 5.0MPa, and the separation temperature is 35 ℃;
(4) static extraction: introducing a certain amount of CO2Raising the pressure in the extraction kettle to the set pressure in the step (3), and firstly, taking the solvent A as an entrainer to perform static extraction for 0.8 h;
(5) dynamic primary extraction: CO regulation2The flow rate of the solvent A is 1.2L/min, and then the solvent A is taken as an entrainer to carry out dynamic primary extraction, wherein the dynamic primary extraction time is 1.5 h;
(6) dynamic secondary extraction: CO regulation2The flow rate of the solvent B is 1.3L/min, and finally, the entrainer pump is used for adding the solvent B into the extraction kettle to be used as the entrainerDynamic secondary extraction is carried out, and the time of the dynamic secondary extraction is 2.5 hours; the solvent B is a mixture of n-butyl alcohol and tert-butyl methyl ether, and the using amount of the solvent B is 60 wt% of the mass of the jujube kernel raw material;
(7) separating in a separation kettle I for 2h and a separation kettle II for 1h, and mixing the jujube kernel volatile oil obtained in the separation kettle I and the separation kettle II;
(8) and (3) separating and purifying the jujube kernel volatile oil obtained in the step (7) by column chromatography, wherein a) the column chromatography: an ACQUITY UPLC BEHC18 column, 2.1mm × 50mm,1.7 μm; b) the mobile phase consists of a phase A and a phase B: the phase A is as follows: methanol, phase B is: 1mmol/L ammonium formate aqueous solution; c) gradient elution conditions: the volume ratio of the phase A to the phase B is 10:1 at the beginning, the volume ratio of the phase A to the phase B is 8:3 at 1 minute, the volume ratio of the phase A to the phase B is 7:4 at 3 minutes, the volume ratio of the phase A to the phase B is 6:5 at 6 minutes, the volume ratio of the phase A to the phase B is 5:6 at 9 minutes, the volume ratio of the phase A to the phase B is 4:7 at 12 minutes, the volume ratio of the phase A to the phase B is 3:8 at 15 minutes, the volume ratio of the phase A to the phase B is 1:10 at 18 minutes, and the collection starting time is as follows: 1min, end of collection time: 18 min; d) column temperature: 35 ℃; e) flow rate: 0.6 mL/min; removing mobile phase to obtain pure Zizyphi Spinosae semen volatile oil.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 22.75%.
Example 4:
the application of jujube kernel volatile oil in the hair growth of mice comprises the following steps:
(1) pre-treating the jujube kernel raw material: oven drying semen Ziziphi Spinosae at 60 deg.C for 23 hr, pulverizing, and sieving to obtain powder with particle size of 50-70 mesh;
(2) charging: adding the pretreated semen Ziziphi Spinosae material and solvent A into supercritical CO2In an extraction kettle of a fluid extraction device; in this example, solvent a is ethyl acetate; the dosage of the solvent A is 25 wt% of the mass of the jujube kernel raw material;
(3) setting the temperature and pressure of the extraction kettle, the separation kettle I and the separation kettle II;
the extraction pressure of the extraction kettle is 32MPa, and the extraction temperature is 45 ℃; the separation pressure of the separation kettle I is 8.3MPa, and the separation temperature is 33 ℃; the separation pressure of the separation kettle II is 5.4MPa, and the separation temperature is 36 ℃;
(4) static extraction: introducing a certain amount of CO2Raising the pressure in the extraction kettle to the set pressure in the step (3), and firstly, taking the solvent A as an entrainer to perform static extraction for 0.7 h;
(5) dynamic primary extraction: CO regulation2The flow rate of the solvent A is 1.4L/min, and then the solvent A is taken as an entrainer to carry out dynamic primary extraction, wherein the dynamic primary extraction time is 1.5 h;
(6) dynamic secondary extraction: CO regulation2The flow rate of the extraction kettle is 1.4L/min, and finally, a solvent B is added into the extraction kettle by an entrainer pump to be used as an entrainer for dynamic secondary extraction, wherein the dynamic secondary extraction time is 2.2 h; the solvent B is a mixture of n-butyl and tert-butyl methyl ether, the volume ratio of n-butyl to tert-butyl methyl ether is 1:3, and the amount of the solvent B is 80 wt% of the mass of the jujube kernel raw material;
(7) separating in separation kettle I for 2.5h and separation kettle II for 1.5h, and mixing the volatile oil obtained in separation kettle I and separation kettle II;
(8) and (3) separating and purifying the jujube kernel volatile oil obtained in the step (7) by column chromatography, wherein a) the column chromatography: an ACQUITY UPLC BEHC18 column, 2.1mm × 50mm,1.7 μm; b) the mobile phase consists of a phase A and a phase B: the phase A is as follows: methanol, phase B is: 1mmol/L ammonium formate aqueous solution; c) gradient elution conditions: the volume ratio of the phase A to the phase B is 10:1 at the beginning, the volume ratio of the phase A to the phase B is 8:3 at 1 minute, the volume ratio of the phase A to the phase B is 7:4 at 3 minutes, the volume ratio of the phase A to the phase B is 6:5 at 6 minutes, the volume ratio of the phase A to the phase B is 5:6 at 9 minutes, the volume ratio of the phase A to the phase B is 4:7 at 12 minutes, the volume ratio of the phase A to the phase B is 3:8 at 15 minutes, the volume ratio of the phase A to the phase B is 1:10 at 18 minutes, and the collection starting time is as follows: 1min, end of collection time: 18 min; d) column temperature: 35 ℃; e) flow rate: 0.6 mL/min; removing mobile phase to obtain pure Zizyphi Spinosae semen volatile oil.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 23.26%.
Example 5:
this example differs from example 1 in that: the step (1) also comprises the steps of sequentially carrying out alkali soaking, acid soaking, buffer solution soaking and air drying on the crushed and sieved jujube kernel raw material; the alkaline solution used for alkaline soaking is a sodium carbonate solution with the mass concentration of 5 wt%, the dosage of the alkaline solution per 100g of jujube kernel powder is 300mL, the alkaline soaking time is 5h, and the filtering is carried out after the alkaline soaking; the acid solution used for acid soaking is hydrochloric acid with the mass concentration of 10 wt%, the dosage of the alkali solution per 100g of jujube kernel powder is 200mL, the acid soaking time is 0.5h, and the filtering is carried out after the acid soaking; the buffer solution used for soaking is phosphoric acid buffer solution with pH 7.2, and the soaking time of the buffer solution is 5 h.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 23.82%.
Example 6:
this example differs from example 2 in that: the step (1) also comprises the steps of sequentially carrying out alkali soaking, acid soaking, buffer solution soaking and air drying on the crushed and sieved jujube kernel raw material; the alkaline solution used for alkaline soaking is a sodium carbonate solution with the mass concentration of 12 wt%, the dosage of the alkaline solution per 100g of jujube kernel powder is 400mL, the alkaline soaking time is 10h, and the filtering is carried out after the alkaline soaking; the acid solution used for acid soaking is hydrochloric acid with the mass concentration of 15 wt%, the dosage of the alkali solution per 100g of jujube kernel powder is 300mL, the acid soaking time is 2h, and the filtering is carried out after the acid soaking; the buffer solution used for soaking is phosphoric acid buffer solution with pH 7.2, and the soaking time of the buffer solution is 10 h.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 24.31%.
Example 7:
this example differs from example 3 in that: the step (1) also comprises the steps of sequentially carrying out alkali soaking, acid soaking, buffer solution soaking and air drying on the crushed and sieved jujube kernel raw material; the alkaline solution used for alkaline soaking is a sodium carbonate solution with the mass concentration of 10 wt%, the dosage of the alkaline solution per 100g of jujube kernel powder is 350mL, the alkaline soaking time is 8h, and the filtering is carried out after the alkaline soaking; the acid solution used for acid soaking is hydrochloric acid with the mass concentration of 12 wt%, the dosage of the alkali solution per 100g of jujube kernel powder is 250mL, the acid soaking time is 1h, and the filtering is carried out after the acid soaking; the buffer solution used for soaking is phosphoric acid buffer solution with pH 7.2, and the soaking time of the buffer solution is 8 h.
The mass of the jujube kernel volatile oil is finally obtained by weighing, and the yield of the obtained jujube kernel volatile oil is 23.71%.
Example 8:
this example differs from example 4 in that: the step (1) also comprises the steps of sequentially carrying out alkali soaking, acid soaking, buffer solution soaking and air drying on the crushed and sieved jujube kernel raw material; the alkaline solution used for alkaline soaking is a sodium carbonate solution with the mass concentration of 7 wt%, the dosage of the alkaline solution per 100g of jujube kernel powder is 360mL, the alkaline soaking time is 9h, and the filtering is carried out after the alkaline soaking; the acid solution used for acid soaking is hydrochloric acid with the mass concentration of 11 wt%, the dosage of the alkali solution per 100g of jujube kernel powder is 300mL, the acid soaking time is 2h, and the filtering is carried out after the acid soaking; the buffer solution used for soaking is phosphoric acid buffer solution with pH 7.2, and the soaking time of the buffer solution is 6 h.
The mass of the jujube kernel volatile oil is finally obtained through weighing, and the yield of the obtained jujube kernel volatile oil is 30.05%.
From examples 5 to 8 it can be seen that: after alkali soaking, acid soaking and buffer solution soaking are sequentially carried out, the yield of the jujube kernel volatile oil is greatly improved, and the total antioxidant capacity of the obtained volatile oil is not greatly changed. However, if the sequence of alkali soaking, acid soaking and buffer solution soaking is changed, the yield of the jujube kernel volatile oil is reduced by more than 26 percent compared with the yield of the jujube kernel volatile oil without soaking.
Examples of promoting Hair growth in mice
The method for promoting the hair growth of the mice comprises the following steps:
(1) selecting a test mouse;
the mice are pink in skin color, and after birth for 40 days, hair grows to be the C57BL/6J inbred line mice in the resting period, and the female and male are used for half and half, 35-65 days, 15-20 g, and 150 mice in total.
(2) Induction of the hair growth period of the mice;
induction of hair growth period: after the mice had undergone a breeding adaptation period of about one week, rosin and paraffin were mixed at a ratio of 1:1, heating and melting the mixture, smearing the mixture on the back of a mouse when the temperature is proper, and quickly tearing the mixture after the mixture is solidified and hardened to achieve the effect of plucking hairs.
(3) Grouping of test mice;
the test was divided into 5 groups of 30 mice each, 3 replicates each, and 10 replicates each (male and female halves). Group 1 was a model blank control group (distilled water was applied after epilation and normal saline was perfused into the stomach); group 2 was a positive control (appliquid 101 and gastric perfusion saline). The remaining 3 groups were experimental groups, in which:
the 3 rd group of external hair growth preparation is composed of glycerol, propylene glycol and jujube kernel volatile oil, the volume ratio of the glycerol to the propylene glycol is 3:4, and the content of the jujube kernel volatile oil in the external hair growth preparation is 50 mg/mL; the administration amount of the jujube kernel volatile oil for intragastric administration of mice is 1g/Kg, namely 1000 mg of the jujube kernel volatile oil for intragastric administration of mice per kilogram; the jujube kernel volatile oil used in this group was the jujube kernel volatile oil prepared in example 1.
The group 4 of the external application hair growth preparation consists of glycerol, propylene glycol and jujube kernel volatile oil, the volume ratio of the glycerol to the propylene glycol is 3:8, and the content of the jujube kernel volatile oil in the external application hair growth preparation is 100 mg/mL; the administration amount of the jujube kernel volatile oil for intragastric administration of mice is 3g/Kg, namely, the administration is performed according to the weight of the mice, and 3000 mg is administered per kilogram of mice; the jujube kernel volatile oil used in this group was the jujube kernel volatile oil prepared in example 2.
The group 5 of the external application hair growth preparation consists of glycerol, propylene glycol and jujube kernel volatile oil, the volume ratio of the glycerol to the propylene glycol is 3:6.2, and the content of the jujube kernel volatile oil in the external application hair growth preparation is 76 mg/mL; the administration amount of the jujube kernel volatile oil by intragastric administration for the mice is 2.3g/Kg, namely, the administration by intragastric administration is carried out according to the weight of the mice, and 2300 mg of the administration is carried out for each kilogram of the mice; the jujube kernel volatile oil used in this group was the jujube kernel volatile oil prepared in example 8.
(4) Smearing semen Ziziphi Spinosae volatile oil;
the mode of smearing the jujube kernel volatile oil on the mouse is as follows: depilating the back of mouse with mild depilatory 24 hr before applying semen Ziziphi Spinosae volatile oil, and depilating the skin with a depilatory area of 4cm2Applied to the plucked area 2-3 times daily, in successionAnd (5) continuously applying for 21d, and completely growing the hairs applied to the back hair removal area.
(5) Observing the growth cycle of the mouse hair;
observation of the mouse hair growth cycle: the skin color change of the mice was observed every day, and the time from the application of the drug to the beginning of blackening, the beginning of growing hair and the beginning of graying of the skin color of the depilated area of the back of each mouse was recorded and periodically photographed. The time from application to the beginning of darkening of the skin was taken as the time from telogen to anagen, and the time from the beginning of darkening to the beginning of graying was taken as the total time of anagen.
(6) Measurement of mouse hair length;
measurement of mouse hair length: the hair length was measured every 3 days from the back growth of the mice until the end of the test. Plucking 3 parts of the back of each group of mice from head to tail, wherein 5-10 hairs are plucked at each part, measuring the plucked hairs by using a vernier caliper, and recording the farthest distance between two ends of the hairs as the length of the hairs.
The effects of the hair growth rate and growth time and growth cycle of the mice were observed in comparison with the blank control group and the positive control group, and the test results are shown in tables 1 and 2:
TABLE 1 Effect of Ziziphus jujuba seed volatile oil on the Hair growth Rate and growth time of C57BL/6J mice ((
n＝10)
Note: comparison with blank control group1)P＜0.01，2)P is less than 0.05, and is compared with a positive control group3)P＜0.01，4)P is less than 0.05 (same as table 2); the positive control group was administered with octopus 101.
TABLE 2 Effect of Ziziphus jujuba seed volatile oil on the Hair growth cycle of C57BL/6J mice ((
n＝10)
As can be seen from Table 1, the jujube kernel volatile oil has the function of accelerating hair growth in the early and middle hair growth stages, shortens the hair growth time, and has significant difference (P is less than 0.01) compared with a blank control group; as can be seen from table 2, after the mice had depilated, the skin appeared pink, the hair entered the resting period, after the application of the drug for 7-10 days, the positive control group and the test group 3 began to blacken, after 12-15 days, the mice in the test groups 1 and 2 began to blacken, after 14-16 days, the blank group began to blacken, and the blank control group, the positive control group and the test group all had significant differences. After applying the medicine for 13-17 d, the black skin of the positive control group and the test group began to grow hair, after 17-19 d, the hair of the test group 1 began to grow, and after 21d, the hair of the blank group began to grow. Each test group was significantly different from the blank control group. The application time of each group is not significantly different because each administration group is full of hair in 26-29 days.
The above examples are provided for clarity of illustration only and are not intended to limit the invention to the particular embodiments described. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any obvious variations or modifications which come within the spirit and scope of the invention are desired to be protected by the following claims.
Claims (1)
1. An application of jujube kernel volatile oil in the hair growth of mice is characterized in that the jujube kernel volatile oil is taken as an active ingredient for promoting the hair growth of the mice, and the administration by gastric gavage and the smearing of the part to be grown are carried out simultaneously;
the stomach-filling administration amount of the jujube kernel volatile oil for the mice is 2.3g/Kg, the part of the mice to be grown with the hair is coated with an external hair growth preparation taking the jujube kernel volatile oil as an active ingredient, the external hair growth preparation consists of glycerol, propylene glycol and the jujube kernel volatile oil, the volume ratio of the glycerol to the propylene glycol is 3:6.2, and the content of the jujube kernel volatile oil in the external hair growth preparation is 76 mg/mL;
the jujube kernel volatile oil is prepared by the following steps:
(1) pre-treating the jujube kernel raw material: oven drying semen Ziziphi Spinosae at 60 deg.C for 23 hr, pulverizing, and sieving to obtain powder with particle size of 50-70 mesh; the step (1) also comprises the steps of sequentially carrying out alkali soaking, acid soaking, buffer solution soaking and air drying on the crushed and sieved jujube kernel raw material; the alkaline solution used for alkaline soaking is a sodium carbonate solution with the mass concentration of 7 wt%, the dosage of the alkaline solution per 100g of jujube kernel powder is 360mL, the alkaline soaking time is 9h, and the filtering is carried out after the alkaline soaking; the acid solution used for acid soaking is hydrochloric acid with the mass concentration of 11 wt%, the dosage of the alkali solution per 100g of jujube kernel powder is 300mL, the acid soaking time is 2h, and the filtering is carried out after the acid soaking; the buffer solution used for soaking is phosphoric acid buffer solution with pH 7.2, and the soaking time of the buffer solution is 6 h;
(2) charging: adding the pretreated semen Ziziphi Spinosae material and solvent A into supercritical CO2In an extraction kettle of a fluid extraction device; in this example, solvent a is ethyl acetate; the dosage of the solvent A is 25 wt% of the mass of the jujube kernel raw material;
(3) setting the temperature and pressure of the extraction kettle, the separation kettle I and the separation kettle II;
the extraction pressure of the extraction kettle is 32MPa, and the extraction temperature is 45 ℃; the separation pressure of the separation kettle I is 8.3MPa, and the separation temperature is 33 ℃; the separation pressure of the separation kettle II is 5.4MPa, and the separation temperature is 36 ℃;
(4) static extraction: introducing a certain amount of CO2Raising the pressure in the extraction kettle to the set pressure in the step (3), and firstly, taking the solvent A as an entrainer to perform static extraction for 0.7 h;
(5) dynamic primary extraction: CO regulation2The flow rate of the solvent A is 1.4L/min, and then the solvent A is taken as an entrainer to carry out dynamic primary extraction, wherein the dynamic primary extraction time is 1.5 h;
(6) dynamic secondary extraction: CO regulation2The flow rate of the extraction kettle is 1.4L/min, and finally, a solvent B is added into the extraction kettle by an entrainer pump to be used as an entrainer for dynamic secondary extraction, wherein the dynamic secondary extraction time is 2.2 h; the solvent B is a mixture of n-butyl and tert-butyl methyl ether, the volume ratio of n-butyl to tert-butyl methyl ether is 1:3, and the amount of the solvent B is 80 wt% of the mass of the jujube kernel raw material;
(7) separating in separation kettle I for 2.5h and separation kettle II for 1.5h, and mixing the volatile oil obtained in separation kettle I and separation kettle II;
(8) and (3) separating and purifying the jujube kernel volatile oil obtained in the step (7) by column chromatography, wherein a) the column chromatography: ACQUITY UPLC BEH C18 column, 2.1mm × 50mm,1.7 μm; b) the mobile phase consists of a phase A and a phase B: the phase A is as follows: methanol, phase B is: 1mmol/L ammonium formate aqueous solution; c) gradient elution conditions: the volume ratio of the phase A to the phase B is 10:1 at the beginning, the volume ratio of the phase A to the phase B is 8:3 at 1 minute, the volume ratio of the phase A to the phase B is 7:4 at 3 minutes, the volume ratio of the phase A to the phase B is 6:5 at 6 minutes, the volume ratio of the phase A to the phase B is 5:6 at 9 minutes, the volume ratio of the phase A to the phase B is 4:7 at 12 minutes, the volume ratio of the phase A to the phase B is 3:8 at 15 minutes, the volume ratio of the phase A to the phase B is 1:10 at 18 minutes, and the collection starting time is as follows: 1min, end of collection time: 18 min; d) column temperature: 35 ℃; e) flow rate: 0.6 mL/min; removing mobile phase to obtain pure Zizyphi Spinosae semen volatile oil.
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| CN101396381A (en) * | 2007-09-29 | 2009-04-01 | 北京北大维信生物科技有限公司 | Valerian extract and preparation method and use thereof |
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