Natural plant acarus killing emulsion as well as preparation method and application thereof

Abstract

The invention discloses a natural plant acarus killing emulsion as well as a preparation method and application thereof. The natural plant mite-killing emulsion is prepared from, by mass, 5%-10% of litsea cubeba volatile oil, 5%-10% of herba ecliptae total saponins, 4%-8% of emulsifier, 3%-5% of co-emulsifier, 15%-25% of triglyceride and the balance water. The components are added with distilled water to complement to 100%, and a uniform emulsion is prepared. The effect of killing dust mites is achieved by utilizing the activity of volatile oil and saponin. And the emulsion can enable the medicinal components to be slowly volatilized, so that a long-acting insecticidal effect is achieved. The product can be used in the fields of washing daily necessities, cosmetics and the like.

Description

Technical Field

[0001] The invention relates to the technical field of preparation of traditional Chinese medicine compositions and mite removal in daily chemical products, and in particular to a natural plant mite removal emulsion containing litsea cubeba volatile oil and eclipta saponin, and a preparation method and application thereof. Background Art

[0002] As we all know, mites are widely distributed in the environment we live in. In damp corners, clothes, fabrics and household items that humans come into contact with on a daily basis may be infested with mites. Long-term contact with mites can easily cause allergies, various skin diseases, etc., causing trouble to human life. Currently used mite removers are mostly chemical mite removers, which are easy to leave residues, and are easy to cause harm to health after long-term contact or inhalation, and are prone to drug resistance. Therefore, it is of great significance to develop green, safe, natural plant-based mite removers.

[0003] Many studies have shown that various ingredients in plant extracts have good repelling and killing effects on mites. However, there are not many types of pure Chinese medicine preparations for killing mites. The disclosed plant-based miticides have complex ingredients and are difficult to control in quality because the raw materials contain many medicinal materials; or some active ingredients are volatile and have a short duration of action. Litsea cubeba volatile oil has antibacterial and mite-killing effects (CN105039016A, CN108103607A, CN101278681A), but the unilateral Litsea cubeba oil has limited mite-killing effect, and evaporates too quickly to achieve a long-term effect. Therefore, it is necessary to develop a natural plant mite-killing preparation with high biosafety and long-term effectiveness. 。 Summary of the invention

[0004] The purpose of the present invention is to provide a natural plant mite removal emulsion and a preparation method and application thereof. The natural plant mite removal emulsion provided by the present invention can effectively kill mites for a long time.

[0005] The first aspect of the present invention provides a natural plant mite removal emulsion, which comprises, by weight percentage: 5%-10% of Litsea cubeba volatile oil, 5%-10% of Eclalphia total saponins, 4%-8% of emulsifier, 3%-5% of auxiliary emulsifier, 15%-20% of triglyceride, and the balance is water.

[0006] Furthermore, the volatile oil of Litsea cubeba is prepared by the following method: fresh fruits of Litsea cubeba are taken, placed in a round-bottom flask, deionized water with a solid-liquid ratio of 1:10~1:20 (W / V) is added, ultrasonic pretreatment is performed for 10 minutes, heating to boiling, and the volatile oil is extracted by steam distillation for 2~4 hours, and the upper oily liquid in the volatile oil extractor is collected to obtain the volatile oil of Litsea cubeba.

[0007] Furthermore, the total saponins of Eclampsia chinensis were prepared by the following method: (1) Extraction: Take the dried whole herb of Eclampsia chinensis, add 8 to 12 times (V / W) of 80% ethanol, reflux extraction twice, each time for 60 minutes, filter, combine the filtrate, and recover the solvent under reduced pressure until there is no alcohol taste to obtain a concentrated solution. (2) Macroporous resin purification: Take the concentrated solution of step (1) and purify it by AB-8 macroporous resin column chromatography. The loading ratio is: V 浓缩液 / W 大孔树脂 =2:1, repeat the loading twice, let it stand for 20 minutes after the loading, first elute with 30% (V / V) ethanol for 2 column volumes and discard; then elute with 80% (V / V) ethanol for 4 column volumes, collect the eluate, recover the solvent, put it into a freeze dryer for freeze drying, and obtain the dry powder of total saponins of Eclaludin.

[0008] Furthermore, the emulsifier is selected from one or more of polysorbate 20 and polysorbate 80; preferably polysorbate 80.

[0009] Furthermore, the co-emulsifier is ethanol with a mass concentration of 70-100%, preferably anhydrous ethanol.

[0010] The second aspect of the present invention provides a method for preparing the above-mentioned natural plant mite removal emulsion, comprising the following specific steps: According to the mass fraction, the volatile oil of Litsea cubeba is dispersed in triglyceride to obtain the oil phase after it is completely dissolved; the total saponins of Eclalaria japonica are added to anhydrous ethanol and dispersed, and then added to distilled water together with the emulsifier polysorbate 80, and the mixture is used as the water phase; at a water bath temperature of 50°C and a stirring speed of 450 rpm, the oil phase is added to the water phase while stirring, so that the oil phase and the water phase are evenly mixed to obtain a uniform emulsion.

[0011] The natural plant mite removal emulsion prepared by the invention has uniform particle size and good stability.

[0012] The natural plant mite removal emulsion of the present invention can be used in the fields of cleaning daily necessities, cosmetics, clothing or environmental mite removal.

[0013] Compared with the prior art, the present invention has at least one of the following beneficial effects: The present invention uses the total saponin formula obtained by extracting and purifying the volatile oil of Litsea cubeba and extracting and purifying it with Eclalphia oleracea. On the one hand, the acaricidal effect of Litsea cubeba oil and the cytotoxic effect of Eclalphia oleracea saponin are fully utilized. On the other hand, Eclalphia oleracea saponin also has a surfactant effect, which promotes the absorption of Litsea cubeba volatile oil by insects. In order to solve the drawbacks of fast volatilization and short efficacy of volatile oil components, the present invention is carefully designed in the dosage form of the two compositions, and the volatile oil of Litsea cubeba is included in the aqueous phase containing Eclalphia oleracea saponin to form an emulsion to achieve a sustained-release long-acting acaricidal effect. In an exemplary embodiment of the present invention, the prepared plant emulsion has a mite killing rate of 80.14% on the first day of use, and a strong mite killing effect is achieved according to the T / CHCIA 002-2022 standard evaluation; and the mite killing rate on the second day after use is 76.09%, and on the third day is 62.92%, which still meets the standard of product effectiveness. It shows that the product can still achieve an effective mite killing effect 3 days after use, which reflects the characteristics of long-term effect compared with other products. The process of the invention is green, environmentally friendly, and can be promoted for use. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 The figures are the 5-day mite killing effects of each group of embodiments and comparative examples. DETAILED DESCRIPTION

[0015] The present invention will be further described below in conjunction with embodiments, but is not limited thereto. Example 1

[0016] Extraction of volatile oil from Litsea cubeba with different extraction solvents: Take fresh Litsea cubeba fruit, place it in a round-bottom flask, add deionized water with a solid-liquid ratio of 1:8~22 (W / V), pre-treat with ultrasound for 10 minutes, heat to boiling, extract volatile oil by steam distillation, the extraction time is 3 hours, collect the upper oily liquid in the volatile oil extractor, and obtain volatile oil from Litsea cubeba. The specific yield of volatile oil from Litsea cubeba under different extraction conditions is shown in Table 1.

[0017] Table 1 Yield of volatile oil from Litsea cubeba under different extraction conditions

[0018] As can be seen from Table 1, when the solid-liquid ratio (W / V) is 1:8, the obtained volatile oil of Litsea cubeba is less, and as the amount of extraction solvent increases, the volume of essential oil extracted also increases. When the solid-liquid ratio (W / V) is 1:20, the yield of volatile oil of Litsea cubeba reaches 3.84, and when the solid-liquid ratio (W / V) ratio of 1:22 is continued to be used for extraction, the yield of volatile oil obtained does not increase much. Therefore, the present invention sets the solid-liquid ratio of the solvent used for extracting volatile oil of Litsea cubeba to 1:10~1:20 (W / V). Example 2

[0019] Extraction of volatile oil from Litsea cubeba at different extraction times: fresh Litsea cubeba fruit was taken and placed in a round-bottom flask, deionized water with a solid-liquid ratio of 1:20 (W / V) was added, ultrasonic pretreatment was performed for 10 minutes, and then heated to boiling. The volatile oil was extracted by steam distillation for 1 to 5 hours, and the upper oily liquid in the volatile oil extractor was collected to obtain the volatile oil from Litsea cubeba. The specific yields of volatile oil from Litsea cubeba at different extraction times are shown in Table 2.

[0020] Table 2 Yield of volatile oil from Litsea cubeba at different extraction times

[0021] As can be seen from Table 2, when the extraction time is 1 h, the yield of the volatile oil of Litsea cubeba is low, and when the extraction time is further increased to 4 h, the yield reaches 3.95, and when the extraction time exceeds 3 h, the yield does not increase much. Considering that the long extraction time has a certain influence on the stability of the volatile oil components and the time cost of extraction, the extraction time of the volatile oil of Litsea cubeba is set to 2-4 hours in the present invention. Example 3

[0022] Effect of extraction solvent on total saponin content of Eclipta prostrata: Take the dried whole herb of Eclipta prostrata, add 6-14 times (V / W) of 70-90% ethanol, reflux extract twice, 60 minutes each time, filter, combine the filtrate, decompress the filtrate to recover the solvent until there is no alcohol taste, and obtain a concentrate. The concentrate is purified by AB-8 macroporous resin column chromatography. The loading ratio is: V 浓缩液 / W 大孔树脂 =2:1, repeat the loading twice, let it stand for 20 minutes after the loading, first elute with 30% (V / V) ethanol for 2 column volumes and discard; then elute with 80% (V / V) ethanol for 4 column volumes, collect the eluate, recover the solvent, put it into a freeze dryer for freeze drying, and obtain the dry powder of total saponins of Eclaludin.

[0023] The total saponin content of the Ecliptaegi extract obtained under specific different ethanol concentrations and volume extraction conditions is shown in Table 3. As can be seen from Table 3, when the amount of extraction solvent is 6 times the medicinal material (V / W), the total saponin content of Ecliptaegi is relatively low compared to other materials. When the amount of solvent is 14 times, the total saponin content is not much different compared to 12 times. Therefore, the present invention sets the amount of solvent used in the extraction of Ecliptaegi to 8 to 12 times (V / W). At the same time, the concentration of ethanol solvent was examined, and it was found that the extraction of 70% ethanol and 90% ethanol was not as good as 80% ethanol, so the ethanol concentration was set to 80%.

[0024] Determination method of total saponin content of Eclipta prostrata: The total saponin content in freeze-dried powder of Eclipta prostrata extract was determined by UV-visible spectrophotometry. Weigh the Eclipta prostrata A reference substance and add methanol to prepare a reference substance solution with a concentration of 0.1 mg / mL. Accurately pipette 0.5 mL, 1.0 mL, 1.5 mL, 2.0 mL, 2.5 mL, and 3.0 mL of the reference substance mother solution respectively, place them in a stoppered test tube, evaporate the solvent in a water bath, add 0.5 mL of 1% vanillin-perchloric acid solution respectively, heat in a 60℃ water bath for 15 minutes, cool with ice water for 5 minutes after taking out, add 5 mL of 70% sulfuric acid, shake well, measure the absorbance at a wavelength of 550 nm, and prepare the standard curve equation with absorbance and concentration. Take an appropriate amount of freeze-dried powder of the test sample to be tested, add methanol to dissolve it, and measure its absorbance at a wavelength of 550 nm in the same way as the reference substance, and substitute it into the standard curve equation to calculate the total saponin content of Eclipta prostrata in the test sample.

[0025] Table 3 Content of total saponins of Eclipta prostrata in each group of samples Example 4

[0026] Effect of elution solvent on total saponin content of Eclipta prostrata: Take the dried whole herb of Eclipta prostrata, add 10 times (V / W) of 80% ethanol, reflux extract twice, each time for 60 minutes, filter, combine the filtrate, decompress the filtrate to recover the solvent until there is no alcohol taste, and obtain a concentrate. The concentrate is purified by AB-8 macroporous resin column chromatography. The loading ratio is: V 浓缩液 / W 大孔树脂 =2:1, repeat loading twice, let stand for 20 minutes after loading, first elute with ethanol with a volume concentration of 30% (V / V) for 2 column volumes, discard; then elute with ethanol with a volume concentration of 70~90% (V / V) for 2~5 column volumes, collect the eluate, recover the solvent, put it into a freeze dryer for freeze drying, and obtain the dry powder of total saponins of Eclipta prostrata. The specific total saponin content obtained under different ethanol elution conditions is shown in Table 4.

[0027] Table 4 Content of total saponins of Eclipta prostrata in each group of samples

[0028] As can be seen from Table 4, when the Ecliptaegi extract was purified by macroporous resin, elution with 70% ethanol may have insufficient solubility, and the amount of total Ecliptaegi saponins obtained was lower than that of elution with 80% ethanol. The amount of total Ecliptaegi saponins obtained by elution with 90% ethanol was also not ideal due to the mixing of impurities such as chlorophyll. Therefore, 80% ethanol was selected as the elution solvent and eluted with 4 column volumes. Example 5

[0029] The natural plant anti-mite emulsion was prepared by using the dried powder of total saponins of Eclipta prostrata obtained by steam distillation for 3 hours with a solid-liquid ratio of 1:20 in Example 1 and the extracted with 10 times (V / W) of 80% ethanol added in Example 3 and eluted with 80% ethanol for 4 column volumes to prepare the natural plant anti-mite emulsion.

[0030] Emulsion formula: 10% volatile oil of Litsea cubeba, 10% total saponins of Eclalphia lappa, 8% polysorbate 80, 5% anhydrous ethanol, 20% triglyceride, and 47% distilled water.

[0031] The preparation method of the emulsion is as follows: according to the formula, the volatile oil of Litsea cubeba is dispersed in triglyceride to obtain an oil phase after the oil phase is completely dissolved; the total saponins of Eclalaria japonica are added to anhydrous ethanol for dispersion, and then added to distilled water together with the emulsifier polysorbate 80, and the mixture is used as the water phase; at a water bath temperature of 50°C and a stirring speed of 450 rpm, the oil phase is added to the water phase while stirring, so that the oil phase and the water phase are evenly mixed to obtain a uniform light green emulsion. Example 6

[0032] The natural plant mite removal emulsion was prepared by using the dried powder of the total saponins of Eclipta prostrata obtained by steam distillation for 3 hours with a solid-liquid ratio of 1:20 in Example 1 and the eluate collected by eluting with 80% ethanol in 12 times the amount (V / W) in Example 3 and eluting with 80% ethanol for 4 column volumes as raw materials. The emulsion formula and preparation method are the same as those in Example 5. Example 7

[0033] The preparation method of this embodiment is the same as that of embodiment 5, except that the emulsion formula is: 5% volatile oil of Litsea cubeba, 10% total saponins of Eclipta prostrata, 6% polysorbate 80, 5% anhydrous ethanol, 16% triglyceride, and 58% distilled water. Example 8

[0034] The preparation method of this embodiment is the same as that of embodiment 5, except that the emulsion formula is: 10% of volatile oil of Litsea cubeba, 5% of total saponins of Eclalphia lappa, 4% of polysorbate 80, 3% of anhydrous ethanol, 25% of triglyceride, and 53% of distilled water. Comparative Example 1

[0035] The preparation method of this comparative example is the same as that of Example 5, except that the emulsion formula is: 10% of Litsea cubeba volatile oil, 8% of polysorbate 80, 5% of anhydrous ethanol, 20% of triglyceride, and 57% of distilled water. Comparative Example 2

[0036] The preparation method of this comparative example is the same as that of Example 5, except that the emulsion formula is: 10% total saponins of Eclalaria japonica, 8% polysorbate 80, 5% anhydrous ethanol, 20% triglyceride, and 57% distilled water. Comparative Example 3

[0037] The preparation method of this comparative example is the same as that of Example 5, except that the raw material for extracting Litsea cubeba is the leaves of Litsea cubeba. Comparative Example 4

[0038] This comparative example is prepared in the same manner as in Example 5, except that the total saponins of Eclalphia lappa were not purified with a macroporous resin. Comparative Example 5

[0039] The preparation method of this comparative example is the same as that of Example 5, except that when the total saponins of Eclalphia lappa were purified by macroporous resin, they were not eluted with 30% (V / V) ethanol for 2 column volumes and then discarded, but were directly eluted with 80% (V / V) ethanol for 4 column volumes, and the eluate was collected, concentrated and dried to obtain the product. Comparative Example 6

[0040] The preparation method of this comparative example is the same as that of Example 5, except that the emulsion formula is: 10% of Litsea cubeba volatile oil, 10% of total saponins of Eclalphia lappa, 8% of polysorbate 80, 20% of triglyceride, and 52% of distilled water.

[0041] Mite removal performance test: The mite removal performance of the emulsions prepared in the examples and comparative examples was tested with reference to the method described in "Evaluation Method for Repellent, Inhibitor and Killer Mites of Daily Chemical Products" (T / CHCIA 002-2022). Take filter paper and spread it flat in a culture dish, drip the sample to be tested on the filter paper to make it completely soaked, and take another filter paper and drip distilled water on it as a blank control. Put a specified number of dust mites and feed in each culture dish and culture them in an incubator at a temperature of 25°C and a relative humidity of 75%. Every 2 hours, observe the death of dust mites, and every 24 hours, count the mortality rate, remove all dust mites, put in a new batch of dust mites, and continue to observe and count. The mite removal results of each group of test samples are shown in Table 5 and Figure 1 According to the T / CHCIA 002-2022 standard, a mite killing rate of ≥60% indicates that the product is effective, ≥80% indicates that the product has a strong mite killing effect, and ≥95% indicates that it has an extremely strong effect. Figure 1 It can be seen that the mite killing rate of Example 1 and Example 4 (80% ethanol, 12 times) exceeded 80% on the first day of use, achieving a strong mite killing effect. After 3 days of use, the mite killing rate was still over 60%, achieving an effective effect of the product, indicating that the product has a mite killing effect and a long action time.

[0042] Table 5 Mite removal effect of each group of samples

[0043] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A natural plant mite removal emulsion, characterized in that: The composition comprises, by weight percentage: 5%-10% of volatile oil of Litsea cubeba, 5%-10% of total saponin of Eclalphia lappa, 4%-8% of emulsifier, 3%-5% of auxiliary emulsifier, 15%-20% of triglyceride, and the balance of water.

2. The natural plant mite removal emulsion according to claim 1, characterized in that: The volatile oil of Litsea cubeba is prepared by the following method: fresh fruits of Litsea cubeba are taken, placed in a round-bottom flask, deionized water with a solid-liquid ratio of 1:10-1:20 (W / V) is added, ultrasonic pretreatment is performed for 10 minutes, heating is performed to boiling, and volatile oil is extracted by steam distillation for 2-4 hours, and the upper oily liquid in the volatile oil extractor is collected to obtain volatile oil of Litsea cubeba.

3. The natural plant mite removal emulsion according to claim 1, characterized in that: The total saponins of Eclalphia lappa are prepared by the following method: (1) Extraction: Take the dried whole herb of Eclipta prostrata, add 8-12 times (V / W) of 80% ethanol, reflux extract twice, each time for 60 minutes, filter, combine the filtrates, and recover the solvent from the filtrates under reduced pressure until there is no alcohol taste to obtain a concentrated solution; (2) Macroporous resin purification: The concentrated solution from step (1) was purified by AB-8 macroporous resin column chromatography. The loading ratio is: V 浓缩液 / W 大孔树脂 =2:1, repeat the loading twice, let it stand for 20 minutes after the loading, first elute with 30% (V / V) ethanol for 2 column volumes and discard; then elute with 80% (V / V) ethanol for 4 column volumes, collect the eluate, recover the solvent, put it into a freeze dryer for freeze drying, and obtain the dry powder of total saponins of Eclaludin.

4. The natural plant mite removal emulsion according to claim 1, characterized in that: The emulsifier is selected from one or more of polysorbate 20 and polysorbate 80.

5. The natural plant mite removal emulsion according to claim 1, characterized in that: The auxiliary emulsifier is ethanol with a mass concentration of 70-100%.

6. The method for preparing the natural plant mite-killing emulsion according to any one of claims 1 to 5, characterized in that: The method comprises the following specific steps: dispersing the volatile oil of Litsea cubeba in triglyceride to obtain an oil phase after the oil phase is completely dissolved; taking the total saponin of Eclalphia lappa and adding it to an emulsifier to disperse it in distilled water, and adding an emulsifier to mix the mixture to obtain the water phase; adding the oil phase to the water phase while stirring at a water bath temperature of 50°C and a stirring speed of 450 rpm to uniformly mix the oil phase and the water phase to obtain the natural plant mite removal emulsion.

7. Use of the natural plant mite removal emulsion according to any one of claims 1 to 5 in the field of cleaning daily necessities, cosmetics, clothing or environmental mite removal.

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