Application of oxytropis fallax as cosolvent and insecticide

Abstract

The invention discloses application of an oxytropis falcata extract as a cosolvent of an organic solvent extract of clerodendrum japonicum, oxytropis falcata contains rich active substances such as triterpenoid saponin and the like, and has good hydrotropy for the organic solvent extract of clerodendrum japonicum, so that the problem that the organic solvent extract of clerodendrum japonicum is insoluble in water is solved to a certain extent; compared with tween-80, the oxytropis fallax extract serving as a cosolvent phase is greener and healthier; the invention further discloses an application of the oxytropis fallax extract and the clerodendrum japonicum organic solvent extract which are matched with each other as an insecticide.

Description

Technical Field

[0001] The invention relates to Oxytropis flavonoids, and in particular to application of Oxytropis flavonoids as a cosolvent and insecticide. Background Art

[0002] Oxytropis ochrocephala Bunge, commonly known as Tuanba grass and horse intestine, is widely distributed in western my country and is one of the most serious poisonous weeds that harm natural grasslands in the west. Oxytropis ochrocephala Bunge contains a variety of active ingredients, such as alkaloids, flavonoids, triterpenoid saponins, etc., which makes it of great research value. Its strong adaptability in the wild makes it a dominant species in the local area, with a distribution area of ​​more than 3 million hm2. The rich resource reserves also provide a material basis for subsequent in-depth research.

[0003] Clerodendrum japonicum, commonly known as Dragon Boat Flower and Champion Red, is a shrub of the Verbenaceae family. It is mainly distributed in Guizhou, Yunnan, Hunan and other places in my country. As a traditional Chinese medicinal material, it not only has a wide range of medicinal value, such as dispelling wind and dampness, dispersing blood stasis and swelling, and clearing away heat and detoxification, but also has received certain attention in the field of biological pesticides. Tian Wen et al. found that some specific compounds in the petroleum ether extract of Clerodendrum japonicum, such as methyl palmitate, linalool and hexahydrofarnesyl acetone, showed excellent insecticidal activity. The extract has strong insecticidal activity against the larvae of Spodoptera litura, with an LC50 of 12.534 mg / mL. With the improvement of people's awareness of food safety and environmental protection, the use of traditional chemical pesticides has been increasingly restricted. As a natural, low-toxic and efficient biological pesticide resource, Clerodendrum japonicum extract has broad application prospects. It can not only effectively prevent and control insect pests on crops, reduce the use of chemical pesticides, and reduce the harm of pesticide residues to the environment and human body, but also promote the sustainable development of agriculture.

[0004] At present, more than 30 compounds have been isolated and identified from the tung oil tree. The research on the tung oil tree compounds mainly focuses on the medium polarity and water-soluble components, and the research on the petroleum ether part with low polarity is relatively small. In addition, the solubility of the tung oil tree petroleum ether extract in water is low, which makes it difficult to prepare the concentration of the drug solution, and the stability of long-term storage is not satisfactory, which is quite inconvenient to use. There are few research reports on the n-butanol extract of the tung oil tree, and its insecticidal activity also needs further exploration. Summary of the invention

[0005] In view of this, one of the objects of the present invention is to provide an Oxytropis chinensis as a cosolvent to solve the technical problem of low solubility of the organic solvent extract of Tungia oxyphylla in water; the second object of the present invention is to provide an application of Oxytropis chinensis as an insecticide to improve the insecticidal activity of the organic solvent extract of Tungia oxyphylla.

[0006] One of the purposes of the present invention is achieved through the following technical solutions:

[0007] Application of Oxytropis tinctoria extract as a co-solvent for organic solvent extract of Thunbergia paniculata.

[0008] Furthermore, the Oxytropis flavescentis extract is the total extract of Oxytropis flavescentis.

[0009] Furthermore, the preparation method of the total extract of Oxytropis erythrorhizon is as follows: mixing Oxytropis erythrorhizon and a hydrochloric acid aqueous solution with a pH value of 2.51 at a solid-liquid ratio of 28-31:1, reflux extraction for 80-100 minutes at a temperature of 80-100°C, filtering the extract while hot to obtain a filtrate, repeating the reflux extraction and filtration multiple times, combining the filtrate and drying to obtain the total extract of Oxytropis erythrorhizon.

[0010] Furthermore, the organic solvent extract of Tung oil tree is a petroleum ether extract of Tung oil tree or a n-butanol extract of Tung oil tree.

[0011] Further, the following steps are included:

[0012] The Oxytropis flavonoides extract and ultrapure water are mixed to obtain an Oxytropis flavonoides extract aqueous solution; the Oxytropis flavonoides extract aqueous solution is mixed with the Thunb.

[0013] Furthermore, the Oxytropis flavescentis extract is the total extract of Oxytropis flavescentis.

[0014] Furthermore, the preparation method of the total extract of Oxytropis erythrorhizon is as follows: mixing Oxytropis erythrorhizon and a hydrochloric acid aqueous solution with a pH value of 2.51 at a solid-liquid ratio of 28-31:1, reflux extraction for 80-100 minutes at a temperature of 80-100°C, filtering the extract while hot to obtain a filtrate, repeating the reflux extraction and filtration multiple times, combining the filtrate and drying to obtain the total extract of Oxytropis erythrorhizon.

[0015] Furthermore, the concentration of the aqueous solution of Oxytropis flavonoides extract is 17.65 mg / mL.

[0016] Furthermore, the organic solvent extract of Achyranthes bidentata is a petroleum ether extract of Achyranthes bidentata or a n-butanol extract of Achyranthes bidentata.

[0017] Furthermore, the aqueous solution of the Oxytropis flavonoides extract is 200-250 mL, and the petroleum ether extract of the Thunb.

[0018] Furthermore, the aqueous solution of the Oxytropis flavonoides extract is 30-60 mL, and the n-butanol extract of the Thunbergia oxyphylla is 1 g.

[0019] The second object of the present invention is achieved through the following technical solutions:

[0020] Application of Oxytropis flavonoides extract and Thunbergia oxyphylla organic solvent extract as insecticide.

[0021] Furthermore, the method for mixing the extract of Oxytropis flavonoides with the organic solvent extract of Thunb. chinensis comprises the following steps:

[0022] The Oxytropis flavonoides extract and ultrapure water are mixed to obtain an Oxytropis flavonoides extract aqueous solution; the Oxytropis flavonoides extract aqueous solution is mixed with the Thunb.

[0023] Furthermore, the Oxytropis flavescentis extract is the total extract of Oxytropis flavescentis.

[0024] Furthermore, the preparation method of the total extract of Oxytropis erythrorhizon is as follows: mixing Oxytropis erythrorhizon and a hydrochloric acid aqueous solution with a pH value of 2.51 at a solid-liquid ratio of 28-31:1, reflux extraction for 80-100 minutes at a temperature of 80-100°C, filtering the extract while hot to obtain a filtrate, repeating the reflux extraction and filtration multiple times, combining the filtrate and drying to obtain the total extract of Oxytropis erythrorhizon.

[0025] Furthermore, the concentration of the aqueous solution of Oxytropis flavonoides extract is 0.3-6 mg / mL and the amount is 30 mL.

[0026] Furthermore, the organic solvent extract of Tung oil tree is a petroleum ether extract of Tung oil tree or a n-butanol extract of Tung oil tree.

[0027] Furthermore, the amount of the Achyranthes bidentata petroleum ether extract or Achyranthes bidentata n-butanol extract is 15 mg respectively.

[0028] The beneficial effects of the present invention are:

[0029] The invention discloses the use of the Oxytropis vulgaris as a solubility enhancer. The Oxytropis vulgaris contains rich active substances such as triterpenoid saponins, and has good solubility for the organic solvent extract of the Achyranthes bidentata, and solves the problem that the organic solvent extract of the Achyranthes bidentata is difficult to dissolve in water to a certain extent. Compared with Tween-80, the use of the Oxytropis vulgaris extract as a solubility enhancer is greener and healthier.

[0030] A small amount of hydrochloric acid is used to adjust the pH value of water during the preparation of the total extract of Oxytropis tinctorius, so that the organic solvent used in the preparation is also reduced, which saves costs and improves the safety of the preparation.

[0031] When the concentration of total extract of Oxytropis flavonoides in water was 17.65 mg / mL, the maximum solubility of petroleum ether extract and n-butanol extract of Achyranthes bidentata were 4.12 mg / mL and 19.61 mg / mL, respectively.

[0032] The invention discloses an application of the octulophora chinensis as an insecticide. When the total extract of the octulophora chinensis and the organic solvent extract of the tung oil tree are matched at a certain concentration, the LC50 values ​​against 3-day-old Spodoptera litura larvae are 1.081 mg / mL and 1.324 mg / mL, respectively. The insecticidal effect is greatly improved compared with that of a single extract. For example, the LC50 value of the petroleum ether extract of the tung oil tree against Spodoptera litura is only 12.534 mg / mL, indicating that the insecticidal activity of the two after being matched is also enhanced.

[0033] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description and claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further described in detail below with reference to the accompanying drawings, in which:

[0035] Figure 1 This is a curve diagram showing the relationship between the logarithmic units of different concentrations and the probability values ​​of the mutual matching activity of Oxytropis flavonoides and Thunbergia oxyphylla petroleum ether extracts against Spodoptera litura larvae in Example 3 of the present invention.

[0036] Figure 2 This is a curve diagram showing the relationship between the logarithmic units of different concentrations and the probability values ​​of the mutual activity of Oxytropis flavonoides and Thunbergia oxyphylla n-butanol extracts against Spodoptera litura larvae in Example 3 of the present invention. DETAILED DESCRIPTION

[0037] The preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. It should be understood that the preferred embodiments are only for illustrating the present invention, rather than for limiting the protection scope of the present invention.

[0038] The yellow flowered thorn bean involved in the present invention was collected from the natural grassland of Huangzhong County, Qinghai Province in September 2022, and was identified as yellow flowered thorn bean by Professor Mo Chonghui of Qinghai University, plant number (Tancj202209), and stored in the Natural Drug Chemistry Laboratory of the School of Ethnic Medicine of Guizhou University for Nationalities; the test Spodoptera litura eggs were purchased from Jiyuan Keyun Biological Co., Ltd. They were raised with cabbage at a constant temperature [(27±2)℃] indoors, and the relative humidity was controlled between 40% and 60%. After the eggs were hatched, healthy 3rd instar larvae of uniform size were selected for testing; Clerodendrum japonicum was collected from Dushan County, Guizhou Province, and was identified as Clerodendrum japonicum of the Verbenaceae family by Kunming Zhifen Biotechnology Co., Ltd. The wax leaf specimens were stored in the Natural Drug Chemistry Laboratory of the School of Ethnic Medicine of Guizhou University for Nationalities (specimen number: KMZFC20190708 2).

[0039] Embodiment 1

[0040] Application of Oxytropis tinctoria extract as a co-solvent for organic solvent extract of Thunbergia paniculata.

[0041] First, extract the total extract of Oxytropis erythrorhizon:

[0042] Weigh 150g of the whole plant of Oxytropis chinensis in full bloom at 80 mesh and add it to a 1L round-bottom flask, add a hydrochloric acid aqueous solution with a pH value of 2.51 to the round-bottom flask, the solid-liquid ratio of Oxytropis chinensis to the hydrochloric acid aqueous solution is 29.5mL / g, place the round-bottom flask in a 90°C water bath for reflux extraction for 87.5min, filter the extract while hot to obtain a filtrate, repeat the reflux extraction and filtration twice, combine the filtrates and spin-dry on a rotary evaporator to obtain the total extract of Oxytropis chinensis, and obtain 46.49g of the total extract of Oxytropis chinensis after measurement, and calculate the extraction rate P of the total extract of Oxytropis chinensis according to the following formula:

[0043]

[0044] Where:

[0045] P is the extraction rate of total extract of Oxytropis flavonoides (%);

[0046] M1 is the total extract mass of Oxytropis flavonoides (g);

[0047] M2 is the total mass of Oxytropis tinctoria (g).

[0048] The total extraction rate of Oxytropis tinctoria extract was calculated to be 31%.

[0049] Then, the solubility of the total extract of Oxytropis erythrorhizon in water was measured:

[0050] 3g of total extract of Oxytropis erythrorhizon was placed in a 250mL beaker, and ultrapure water was added to the beaker several times at room temperature and stirred. When the volume of ultrapure water was 170mL, there was no suspended matter in the solution, indicating that the total extract of Oxytropis erythrorhizon had been completely dissolved in the ultrapure water and reached a saturated state. At this time, the maximum solubility of the total extract of Oxytropis erythrorhizon in ultrapure water was measured to be 17.65mg / mL. The solubility measurement results of the total extract of Oxytropis erythrorhizon are shown in Table 1:

[0051] Table 1 Solubility measurement results of total extract of Oxytropis tinctoria

[0052]

[0053] Finally, the solubilizing effect of the total extract of Oxytropis chinensis on the petroleum ether extract of Thunbergia paniculata was measured:

[0054] 1g of the petroleum ether extract of Tung Blossom was placed in a 250mL beaker, and 243mL of an aqueous solution of 17.65mg / mL of total extract of Oxytropis chinensis was gradually added to the beaker at room temperature and stirred thoroughly, so that the petroleum ether extract of Tung Blossom was fully dissolved, and there was no suspended matter in the solution, and the saturation state was reached. At this time, the maximum solubility of the petroleum ether extract of Tung Blossom was measured to be 4.12mg / mL. The solubility measurement results of the petroleum ether extract of Tung Blossom when the total extract of Oxytropis chinensis was used as the solubilizing agent are shown in Table 2:

[0055] Table 2 Solubility measurement results of Oxytropis chinensis total extract with petroleum ether dissolution

[0056]

[0057] Embodiment 2

[0058] Application of Oxytropis flavonoides extract as a solvent for n-butanol extract of Thunbergia paniculata.

[0059] First, extract the total extract of Oxytropis erythrorhizon:

[0060] Weigh 150 g of the whole plant of Oxytropis chinensis in full flowering stage with a mesh size of 80 and add it to a 1 L round-bottom flask, add a hydrochloric acid aqueous solution with a pH value of 2.51 to the round-bottom flask, the solid-liquid ratio of Oxytropis chinensis to the hydrochloric acid aqueous solution is 28 mL / g, place the round-bottom flask in a 100° C. water bath for reflux extraction for 80 min, filter the extract while hot to obtain a filtrate, repeat the reflux extraction and filtration twice, combine the filtrates and spin-dry on a rotary evaporator to obtain the total extract of Oxytropis chinensis.

[0061] Then, the solubility of the total extract of Oxytropis chinensis in water was measured:

[0062] 3g of total extract of Oxytropis erythrorhizon was placed in a 250mL beaker, and ultrapure water was added to the beaker several times at room temperature and stirred. When the volume of ultrapure water was 170mL, there was no suspended matter in the solution, indicating that the total extract of Oxytropis erythrorhizon had been completely dissolved in the ultrapure water and reached a saturated state. At this time, the maximum solubility of the total extract of Oxytropis erythrorhizon in ultrapure water was measured to be 17.65mg / mL. The solubility measurement results of the total extract of Oxytropis erythrorhizon are shown in Table 1:

[0063] Table 1 Solubility measurement results of total extract of Oxytropis tinctoria

[0064]

[0065] Finally, the solubilizing effect of the total extract of Oxytropis chinensis on the n-butanol extract of Thunbergia paniculata was measured:

[0066] 1g of the n-butanol extract of Tung Blossom was placed in a 250mL beaker, and 51mL of an aqueous solution of total extract of Oxytropis erythrorhizon with a concentration of 17.65mg / mL was gradually added to the beaker at room temperature and stirred thoroughly, so that the n-butanol extract of Tung Blossom was fully dissolved, and there was no suspended matter in the solution, and the saturation state was reached. At this time, the maximum solubility of the n-butanol extract of Tung Blossom was measured to be 19.61mg / mL. The solubility measurement results of the n-butanol extract of Tung Blossom when the total extract of Oxytropis erythrorhizon was used as the solubility aid are shown in Table 3:

[0067] Table 3 The solubility measurement results of the n-butanol extract of Oxytropis chinensis when the total extract of Oxytropis chinensis was dissolved

[0068]

[0069] Embodiment 3

[0070] Application of Oxytropis flavonoides extract and Thunbergia oxyphylla petroleum ether extract as insecticides.

[0071] First, extract the total extract of Oxytropis erythrorhizon:

[0072] Weigh 150 g of the whole plant of Oxytropis chinensis in full flowering period with a mesh size of 80 and add it to a 1 L round-bottom flask, add a hydrochloric acid aqueous solution with a pH value of 2.51 to the round-bottom flask, the solid-liquid ratio of Oxytropis chinensis to the hydrochloric acid aqueous solution is 31 mL / g, place the round-bottom flask in an 80° C. water bath for reflux extraction for 100 min, filter the extract while hot to obtain a filtrate, repeat the reflux extraction and filtration twice, combine the filtrates and spin-dry on a rotary evaporator to obtain the total extract of Oxytropis chinensis.

[0073] Then, the anti-spodoptera litura activity of the total extract of Oxytropis lutea was tested:

[0074] The insecticidal activity of total extract of Oxytropis chinensis against Spodoptera litura was tested by the insect and leaf immersion method. Weigh 180 mg of total extract of Oxytropis chinensis and add 30 mL of ultrapure water to prepare a 6 mg / mL aqueous solution of total extract of Oxytropis chinensis. Take 90 healthy 3rd-instar Spodoptera litura of the same size as a group. Use a hydrochloric acid aqueous solution with a pH value of 2.51 as a control. The death standard is that the insect body is shriveled and there is no obvious autonomous reaction when lightly touched, and the number of larvae dead at 24, 48, and 72 hours is recorded.

[0075] Since the mortality rate of the control group is between 5% and 20%, it is necessary to correct the mortality rate. The mortality rate and corrected mortality rate are calculated according to the following formula based on the test results.

[0076]

[0077] The results of the activity of the total extract of Oxytropis lutea against the larvae of Spodoptera litura are shown in Table 4.

[0078] Table 4 Activity of total extract of Oxytropis lutea against larvae of Spodoptera litura

[0079]

[0080] From the table above, we can see that the corrected mortality rate is on the rise with the increase of time. The corrected mortality rate of insects immersed in the insect extract for 72 hours is the highest, which is 23.4%.

[0081] Finally, 5 portions of total extract of Oxytropis chinensis were weighed, and 30 mL of ultrapure water was added to prepare aqueous solutions of Oxytropis chinensis with concentrations of 0.375, 0.75, 1.5, 3, and 6 mg / mL, respectively. 15 mg of petroleum ether extract of Tungia chinensis was added to each concentration of aqueous solution of Oxytropis chinensis, and 90 healthy 3rd-instar Spodoptera litura of the same size were taken as a group, and a hydrochloric acid aqueous solution with a pH value of 2.51 was used as a control. The criterion for death was that the insect body was shrunken and had no obvious autonomous reaction when lightly touched, and the number of larval deaths at 24, 48, and 72 hours was recorded.

[0082] Since the mortality rate of the control group is between 5% and 20%, it is necessary to correct the mortality rate. The mortality rate and corrected mortality rate are calculated according to the following formula based on the test results.

[0083]

[0084] The mutual activity of the total extract of Oxytropis lutea and the petroleum ether extract of Thunbergia paniculata against the larvae of Spodoptera litura is shown in Table 5:

[0085] Table 5 The mutual activity of the total extract of Oxytropis chinensis and the petroleum ether extract of Achyranthes bidentata against the larvae of Spodoptera litura

[0086]

[0087] The data showed that the time of immersion in insect solution was positively correlated with the corrected mortality rate. The longer the time, the higher the corrected mortality rate. And with the increase of concentration, the corrected mortality rate also showed an upward trend. When the concentration was 6 mg / mL, the insecticidal activity was the strongest at 72 hours, and the corrected mortality rate of the petroleum ether extract of the tung oil was 57.7%. Compared with the 6 mg / mL total extract of schizonepeta eryngii, the corrected mortality rate at 72 hours was significantly improved, indicating that adding a certain amount of active part extract of tung oil to the total extract solution of schizonepeta eryngii at the same concentration can improve the insecticidal activity.

[0088] Determination of median lethal concentration of Oxytropis chinensis and Thunbergia paniculata petroleum ether extract after mixing:

[0089] SPSS20.0 software was used for data statistical analysis to calculate the median lethal concentration (LC50) value and 95% confidence limit.

[0090] The logarithm of different total alkaloid concentrations was taken as the horizontal axis, and the corrected mortality rate was processed into probability values ​​as the vertical axis. Figure 1As shown, the probability unit model equation of the oil ether extract of Tung oil is: P = 0.21362x-0.4924 (R2 = 0.999). Finally, it was concluded that the LC50 of the oil ether extract of Tung oil to the 3-day-old larvae of Spodoptera litura was 1.081 mg / mL, and the 95% confidence interval was 0.523-1.795. Compared with the LC50 (12.534 mg / mL) of the single oil ether extract of Tung oil to the Spodoptera litura, it was greatly improved. It further illustrates that the combination of the total extract of Oxytropis lutea and the oil ether extract of Tung oil can enhance the insecticidal activity.

[0091] Embodiment 4

[0092] Application of Oxytropis flavonoides extract and Thunbergia oxyphylla n-butanol extract as insecticides.

[0093] First, extract the total extract of Oxytropis erythrorhizon:

[0094] Weigh 150 g of the whole plant of Oxytropis chinensis in full flowering stage at 80 mesh and add it into a 1L round-bottom flask, add a hydrochloric acid aqueous solution with a pH value of 2.51 into the round-bottom flask, the solid-liquid ratio of Oxytropis chinensis to the hydrochloric acid aqueous solution is 28.6 mL / g, place the round-bottom flask in an 85°C water bath for reflux extraction for 95 minutes, filter the extract while hot to obtain a filtrate, repeat the reflux extraction and filtration twice, combine the filtrate, and then spin-dry on a rotary evaporator to obtain the total extract of Oxytropis chinensis.

[0095] Then, the anti-spodoptera litura activity of the total extract of Oxytropis lutea was tested:

[0096] The insecticidal activity of total extract of Oxytropis chinensis against Spodoptera litura was tested by the insect and leaf immersion method. Weigh 180 mg of total extract of Oxytropis chinensis and add 30 mL of ultrapure water to prepare a 6 mg / mL aqueous solution of total extract of Oxytropis chinensis. Take 90 healthy 3rd-instar Spodoptera litura of the same size as a group. Use a hydrochloric acid aqueous solution with a pH value of 2.51 as a control. The death standard is that the insect body is shriveled and there is no obvious autonomous reaction when lightly touched, and the number of larvae dead at 24, 48, and 72 hours is recorded.

[0097] Since the mortality rate of the control group is between 5% and 20%, it is necessary to correct the mortality rate. The mortality rate and corrected mortality rate are calculated according to the following formula based on the test results.

[0098]

[0099] The results of the activity of the total extract of Oxytropis lutea against the larvae of Spodoptera litura are shown in Table 4.

[0100] Table 4 Results of the activity of total extract of Oxytropis lutea against larvae of Spodoptera litura

[0101]

[0102] From the table above, we can see that the corrected mortality rate is on the rise with the increase of time. The corrected mortality rate of insects immersed in the insect extract for 72 hours is the highest, which is 23.4%.

[0103] Finally, 5 portions of total extract of Oxytropis chinensis were weighed, and 30 mL of ultrapure water was added to prepare aqueous solutions of Oxytropis chinensis with concentrations of 0.375, 0.75, 1.5, 3, and 6 mg / mL, respectively. 15 mg of n-butanol extract of Tungia chinensis was added to each concentration of aqueous solution of Oxytropis chinensis, and 90 healthy 3rd-instar Spodoptera litura of the same size were taken as a group, and a hydrochloric acid aqueous solution with a pH value of 2.51 was used as a control. The death standard was that the insect body was shrunken and had no obvious autonomous reaction when lightly touched, and the number of larval deaths at 24, 48, and 72 hours was recorded.

[0104] Since the mortality rate of the control group is between 5% and 20%, it is necessary to correct the mortality rate. The mortality rate and corrected mortality rate are calculated according to the following formula based on the test results.

[0105]

[0106] The results of the mutual activity of the total extract of Oxytropis chinensis and the n-butanol extract of Atractylodes chinensis against the larvae of Spodoptera litura are shown in Table 6: Table 6 The results of the mutual activity of the total extract of Oxytropis chinensis and the n-butanol extract of Atractylodes chinensis against the larvae of Spodoptera litura

[0107]

[0108] The data showed that the time of immersion in insect solution was positively correlated with the corrected mortality rate. The longer the time, the higher the corrected mortality rate. And with the increase of concentration, the corrected mortality rate also showed an upward trend. When the concentration was 6 mg / mL, the insecticidal activity was the strongest at 72 hours, and the corrected mortality rate of the n-butanol extract of the scutellaria aviculare was 61.6%. Compared with the 6 mg / mL total extract of Oxytropis chinensis, the corrected mortality rate at 72 hours was significantly improved, indicating that adding a certain amount of active part extract of the scutellaria aviculare to the total extract solution of the same concentration of the scutellaria aviculare can improve the insecticidal activity.

[0109] Determination of median lethal concentration of Oxytropis chinensis and Thunbergia paniculata n-butanol extract after mixing:

[0110] SPSS20.0 software was used for data statistical analysis to calculate the median lethal concentration (LC50) value and 95% confidence limit.

[0111] The logarithm of different total alkaloid concentrations was taken as the horizontal axis, and the corrected mortality rate was processed into probability values ​​as the vertical axis, such as Figure 2As shown, the probability unit model equation of the n-butanol extract of Tung Blossom was: P = 0.47129x-0.2495 (R2 = 0.999). Finally, it was concluded that the LC50 of the n-butanol extract of Tung Blossom against the 3-day-old larvae of Spodoptera litura was 1.324 mg / mL, and the 95% confidence interval was 0.792-2.091. Compared with the single n-butanol extract of Tung Blossom against the LC50 of Spodoptera litura (20.72 mg / mL), it was greatly improved. It further illustrates that the combination of the total extract of Oxytropis lutea and the organic solvent extract of Tung Blossom can enhance the insecticidal activity.

[0112] The total extract of Oxytropis chinensis with a concentration of 17.65 mg / mL was used to dissolve the active part of Tungia chinensis, and the maximum solubility of the petroleum ether extract and the n-butanol extract of Tungia chinensis were 4.12 mg / mL and 19.61 mg / mL, respectively, indicating that some active substances in the total extract of Oxytropis chinensis have good solubility for the active part of Tungia chinensis. To a certain extent, the problem that the active part of Tungia chinensis is difficult to dissolve in water was solved. And the total extract of Oxytropis chinensis and the organic solvent extract of Tungia chinensis were matched at a certain concentration to the 3-day-old larvae of Spodoptera litura, and the LC50 were 1.081 mg / mL and 1.324 mg / mL, respectively. Compared with the single insecticidal effect, it is also greatly improved. It shows that the insecticidal activity of the two after matching is also enhanced. Compared with Tween-80, the total extract of Oxytropis chinensis in this experiment is more green and healthy. In the process of extracting Oxytropis ovata, a small amount of hydrochloric acid is used to adjust the pH value of water, which reduces the amount of organic solvents used in the extraction process, saving costs and improving the safety of the experiment. This further broadens the use of Oxytropis ovata and provides a scientific basis for the development of new green pesticides.

[0113] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution, which should be included in the scope of the claims of the present invention.

Claims

1. Application of Oxytropis tinctoria extract as a co-solvent for organic solvent extract of Thunbergia paniculata.

2. The use according to claim 1, characterized in that: The octulostridium perfringens extract is the octulostridium perfringens total extract. The preparation method of the octulostridium perfringens total extract is as follows: mixing octulostridium perfringens and a hydrochloric acid aqueous solution with a pH value of 2.51 at a solid-liquid ratio of 28-31:1, reflux extraction for 80-100 minutes at a temperature of 80-100°C, filtering the extract while hot to obtain a filtrate, repeating the reflux extraction and filtration for multiple times, and combining the filtrates and drying to obtain the octulostridium perfringens total extract.

3. The use according to claim 1 or 2, characterized in that: The organic solvent extract of Tung oil tree is petroleum ether extract of Tung oil tree or n-butanol extract of Tung oil tree.

4. A method for dissolving an organic solvent extract of Thunbergia paniculata using an extract of Oxytropis erythrorhizon, characterized in that: The steps include: The Oxytropis flavonoides extract and ultrapure water are mixed to obtain an Oxytropis flavonoides extract aqueous solution; the Oxytropis flavonoides extract aqueous solution is mixed with the Thunb.

5. The method for dissolving the organic solvent extract of Thunbergia paniculata using the extract of Oxytropis flavonoides according to claim 4, characterized in that: The octulostridium perfringens extract is the octulostridium perfringens total extract. The preparation method of the octulostridium perfringens total extract is as follows: mixing octulostridium perfringens and a hydrochloric acid aqueous solution with a pH value of 2.51 at a solid-liquid ratio of 28-31:1, reflux extraction for 80-100 minutes at a temperature of 80-100°C, filtering the extract while hot to obtain a filtrate, repeating the reflux extraction and filtration for multiple times, and combining the filtrates and drying to obtain the octulostridium perfringens total extract.

6. The method for dissolving the organic solvent extract of Achyranthes bidentata by using the extract of Oxytropis flavonoides according to claim 4, characterized in that: The concentration of the aqueous solution of the Oxytropis chinensis extract is 17.65 mg / mL; the organic solvent extract of the Achyranthes bidentata is a petroleum ether extract of the Achyranthes bidentata or a n-butanol extract of the Achyranthes bidentata; when the organic solvent extract of the Achyranthes bidentata is a petroleum ether extract of the Achyranthes bidentata, the aqueous solution of the Oxytropis chinensis extract is 200-250 mL, and the petroleum ether extract of the Achyranthes bidentata is 1 g; when the organic solvent extract of the Achyranthes bidentata is a n-butanol extract of the Achyranthes bidentata, the aqueous solution of the Oxytropis chinensis extract is 30-60 mL, and the n-butanol extract of the Achyranthes bidentata is 1 g.

7. The application of the combination of Oxytropis chinensis extract and the organic solvent extract of Thunbergia paniculata as insecticide.

8. The use according to claim 7, characterized in that: The method for mixing an extract of Oxytropis tinctorius with an organic solvent extract of Thunbergia paniculata comprises the following steps: The Oxytropis flavonoides extract and ultrapure water are mixed to obtain an Oxytropis flavonoides extract aqueous solution; the Oxytropis flavonoides extract aqueous solution is mixed with the Thunb.

9. The use according to claim 8, characterized in that: The octulostridium perfringens extract is the octulostridium perfringens total extract. The preparation method of the octulostridium perfringens total extract is as follows: mixing octulostridium perfringens and a hydrochloric acid aqueous solution with a pH value of 2.51 at a solid-liquid ratio of 28-31:1, reflux extraction for 80-100 minutes at a temperature of 80-100°C, filtering the extract while hot to obtain a filtrate, repeating the reflux extraction and filtration for multiple times, and combining the filtrates and drying to obtain the octulostridium perfringens total extract.

10. The use according to claim 8, characterized in that: The concentration of the aqueous solution of the Oxytropis flavonoides extract is 0.3-6 mg / mL and the amount is 30 mL; the organic solvent extract of the Achyranthes bidentata is a petroleum ether extract of the Achyranthes bidentata or a n-butanol extract of the Achyranthes bidentata; the petroleum ether extract of the Achyranthes bidentata or the n-butanol extract of the Achyranthes bidentata are 15 mg respectively.

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