CN110452170B - Isoquinoline alkaloid compound and preparation method and application thereof - Google Patents
Isoquinoline alkaloid compound and preparation method and application thereof Download PDFInfo
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- A01N43/40—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom six-membered rings
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Abstract
The invention discloses an isoquinoline alkaloid compound and a preparation method and application thereof. The compound is obtained by taking the whole grass of thalictrum amazonicum as a raw material and performing ethyl acetate extraction, silica gel column chromatography and high pressure liquid chromatography separation and purification. The molecular formula is C 12 H 11 NO 2 Having the following structural formula:
the compound was named: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone. The compound has an inhibiting effect on phytophthora nicotianae which is obviously better than that of agricultural streptomycin, and the control effect on the black shank of tobacco reaches (72.2 +/-3.3)%. The invention has the advantages of wide distribution of production raw materials, large biological yield, high alkaloid content, simple compound preparation process, low production cost and easy realization of industrial application.
Description
Technical Field
The invention belongs to the technical field of biological pesticides, and particularly relates to an isoquinoline alkaloid compound extracted and separated from thalictrum amazonicum for the first time. Meanwhile, the invention also relates to a preparation method of the compound and application of the compound in prevention and treatment of tobacco black shank.
Background
Thalictrum L.belonging to Ranunculaceae is a perennial herb. There are about 200 plants in the world, and about 67 plants in China, most of which are distributed in the southwest. In plants of genus thalictrum worldwide, there are more than 90 species of the genus plants reported as chemical components, and the main active components are alkaloid, saponin, flavone, etc. Over 43 plants of this genus have folk medicinal records in china. Thalictrum aquilegifolium (Latin's name: thalictrum minus) is a herbaceous plant of Thalictrum, the plant is completely hairless, the stem height is 28-55 cm, the stem grows four times and three times and produces feather-shaped compound leaves. The inflorescence is conical; sepals 4, light yellow-green; most stamens, filaments. The emaciation fruit is narrow, oval, spherical and slightly flat. Flowering in 6-7 months. The altitude is 1400-2700 m, and the plants are in grass slope, field side, irrigation bush or forest. Widely distributed in Asia and Europe. Is mainly distributed in the areas of Sichuan, yunnan West, qinghai, xinjiang, gansu, shanxi, and the like. The plant is a common Chinese medicinal material used in Yunnan and Sichuan folks, and can be used for treating dysentery and diarrhea. Early-stage phytochemical research at home and abroad shows that the thalictrum suborioides is rich in alkaloid active ingredients.
The tobacco black shank is one of the most devastating diseases in tobacco production, and is also called tobacco epidemic disease, and the tobacco growers are called black stalk crazy, black root, aconite disease and the like. The main tobacco producing areas in China occur in different degrees, wherein Anhui, shandong and Henan provinces are historically serious disease areas; the occurrence of tobacco in southern areas such as Yunnan, guizhou, sichuan, hunan, guangdong, guangxi and Fujian is also quite common. At present, the control of the black shank is mainly realized by methods such as crop rotation cultivation, variety gene improvement, biological pesticide and the like. Where control with biopesticides is the most common and most easily achieved method.
The discovery of new active substances with plant disease control efficacy from medicinal plant resources has been a hot point of biopesticide research at home and abroad. According to the invention, a novel isoquinoline alkaloid compound is obtained by research and activity screening of chemical components of thalictrum amazonicum, and extraction and separation, and the compound has no related report so far, and is worth mentioning that the compound has a remarkable effect of resisting the activity of tobacco black shank virus.
Disclosure of Invention
The invention aims to provide a novel isoquinoline alkaloid compound.
Another object of the present invention is to provide a method for preparing said isoquinoline alkaloid compounds.
The invention also aims to provide application of the isoquinoline alkaloid compound in prevention and control of tobacco black shank.
The purpose of the invention is realized by the following technical scheme.
All percentages used in the present invention are mass percentages unless otherwise indicated.
An isoquinoline alkaloid compound with molecular formula of C 12 H 11 NO 2 Having the following structural formula:
the compound was named: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone, english name: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone.
A method of preparing said isoquinoline alkaloid compounds comprising the steps of:
(1) Extracting the extractum: drying the whole plant of thalictrum amazonicum, crushing to 35-60 meshes, placing the crushed sample in a glass reaction kettle, performing reflux extraction for 2 times by using 95% ethanol, combining the two extracting solutions, concentrating to a small volume, diluting by using 3% tartaric acid solution, and extracting for 2 times by using ethyl acetate; saturating the water phase with sodium carbonate, extracting with ethyl acetate for 2 times, mixing the ethyl acetate phases, and concentrating under reduced pressure to obtain alkaloid extract;
(2) Silica gel column chromatography: dissolving the extract obtained in the step (1) by using 1.5-3 times of pure methanol or pure ethanol or pure acetone, mixing the extract with 0.8-2.5 times of 80-100 meshes of silica gel, and drying; silica gel filled in the column is 150-200 meshes, and the dosage of the silica gel is 3-6 times of the weight of the extract; performing gradient elution with chloroform-acetone solutions at a weight ratio of 20;
(3) High-pressure liquid chromatography separation and purification: and (3) taking 6 parts of the column chromatography gradient eluent for high-pressure liquid chromatography separation and purification: using Zorbax PrepHT GF of Agilent company, a 21.2mm multiplied by 25cm reverse phase column, using 40% methanol water solution as a mobile phase, the flow rate is 20mL/min, the detection wavelength of an ultraviolet detector is 292nm, collecting a chromatographic peak of 22.7min, accumulating for multiple times, and evaporating to dryness to obtain the isoquinoline alkaloid compound.
And (3) dissolving the compound obtained after the separation and purification by the high pressure liquid chromatography in pure methanol, and separating by a sephadex column chromatography with the pure methanol as a mobile phase to obtain a yellow jelly, namely a pure compound.
Structural identification of the compounds:
the compound of the invention is yellow jelly, and HRESI-MS shows that the excimer ion peak is 224.0680 2 [ M ] +Na] + Is combined with 1 H and 13 c NMR spectrum to confirm the molecular formula as C 12 H 11 NO 2 Being alkaloid compounds. The infrared spectrum of the compound showed that the compound had a hydroxyl group (3389 cm) -1 ) Carbonyl group (1668 cm) -1 ) And aromatic rings (1615, 1547 and 1442 cm) -1 ) The signals, UV spectra, have absorption maxima at 212, 258, 292, and 334nm, also confirming the presence of aromatic ring structures in the compounds. Process for preparing compounds 1 H、 13 C-NMR and DEPT spectra (Table-1) showed that it contained 12 carbons and 11 hydrogens. The method comprises the following steps: a 1,6, 7-substituted isoquinoline core (C-1 to C-10H-3, H-4, H-5, and H-8), an acetyl group (-CO-CH 3 (ii) a C-1 'and C-2'; h 3 -2 '), one methyl group (C-3'; h 3 -2') and a phenolic hydroxyl group (. Delta.)) H 10.75 s). The isoquinoline nucleus can exist through H-3 and C-1, C-4, C-10; h-4 and C-3, C-5, C-9, C-10; h-5 and C-4, C-9, C-10; and HMBC correlation of H-8 and C-1, C-9, C-10 (FIG. 3).
After the parent nucleus of the compound is determined, the positions of the remaining substituents (acetyl, methyl and phenolic hydroxyl) can be determined by further analysis of its HMBC correlation spectrum. By acetylmethyl hydrogen H 3 -2′(δ H 2.46 And C-1 (. Delta.) C 156.5 HMBC correlation, it was determined that the acetyl substitution was at C-1; by aromatic ring methyl hydrogen H 3 -3′(δ H 2.30 And C-6 (. Delta.) C 160.1)、C-7(δ C 154.0 And C-8 (. Delta.) C 127.5 And H-8 (. Delta.) H 8.49 C-3' (delta) C 15.2 HMBC correlation of) can determine that the methyl substitution is at the C-7 position; the phenolic hydroxyl group can be substituted at the C-6 position by a phenolic hydroxyl hydrogen (delta) H 10.75 And C-5 (. Delta.) C 112.4)、C-6(δ C 160.1)、 C-7(δ C 132.2 ) was confirmed. The structure of this compound was confirmed and the compound was named: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone.
TABLE-1 preparation of the compounds 1 H and 13 c NMR data (C) 5 D 5 N,500 and 125 MHz)
Infrared, ultraviolet and mass spectral data of compounds: UV (CH) 3 OH)λ max (logε)212(4.04)、258(3.36)、292(3.15)、334(3.32)nm；IR(KBr)ν max 3389、3060、2961、 1668、1615、1547、1442、1376、1242、1168、1072、938、826cm -1 ； 13 C-NMR and 1 H-NMR data (C) 5 D 5 N,125 and 500 MHz), see table-1; ESIMS M/z 224 [ M + Na ]] + ；HRESIMS m/z 224.0680[M+Na] + (calculation C) 12 H 11 NNaO 2 ， 224.0687)。
The isoquinoline alkaloid compound is applied to the prevention and treatment of tobacco black shank.
1. The tobacco black shank is caused by phytophthora nicotianae infection, and the ability of the compound of the invention to inhibit the activity of phytophthora is determined, which comprises the following steps:
(1) Preparation of oatmeal agar culture medium: adding 1000mL of water into oatmeal, heating for 1 hour in a boiling water bath, filtering with gauze, adding water to 1000mL, adding sugar and agar, heating to completely melt the agar, filtering with gauze (absorbent cotton in the middle), sterilizing at 121 deg.C for 20min at 15 pounds, cooling to about 45 deg.C, adding ampicillin (5 mg/100 mL) on a sterile operating platform, mixing, pouring into a flat dish, culturing at 28 deg.C for 48 hours, and checking for sterility.
(2) Bacteriostatic experiments: circular filter paper with diameter of 5mm is placed in a culture dish, sterilized under high pressure for 30min under 15 pounds, and then is respectively immersed in 20 mu M compound, 75% ethanol solution and sterilized distilled water after being dried. Respectively sucking 0.2mL of fresh bacterial liquid of phytophthora nicotianae on oatmeal agar medium plates by using a sterile suction pipe on a sterile operating table. Coating with triangular glass coating rod, lightly sticking the filter paper sheets onto corresponding flat plates with tweezers, culturing at 28 deg.C, observing experimental results, and measuring the size of antibacterial zone. Meanwhile, agricultural streptomycin is used as a positive control.
The test result shows that: the diameter of the inhibition zone of the isoquinoline alkaloid compound is 14.6 +/-1.2 mm, and the diameter of the inhibition zone of the positive control agricultural streptomycin is 12.2 +/-1.0 mm. The compound has the effect of inhibiting phytophthora nicotianae remarkably superior to that of positive control agricultural streptomycin and has outstanding activity of inhibiting black shank.
2. The compound of the invention has the following effects of preventing and treating tobacco black shank:
transplanting tobacco seedlings into flowerpots with the diameter of 10cm and the height of 10cm, wherein the culture medium is as follows: sterilized soil, peat, perlite culture (2. After transplanting and seedling slowing, 10g of bacterial grains are added into roots, tobacco seedlings are placed in an artificial climate room for cultivation, the temperature is 30 ℃ in the daytime, the temperature is 28 ℃ in the dark at night, the relative humidity is 95%, and the disease attack of the tobacco seedlings is caused. Before the black shank disease occurs, the tobacco seedlings are subjected to root irrigation treatment by using 20 mu M of the compound, and 10mL of the compound is irrigated to each tobacco seedling; the number of the plants is 2, 10 plants are treated, the treatment is repeated for 3 times, and after 14 days, the disease condition is investigated and the disease index is calculated. The results show that: the compound has a control effect on tobacco black shank of (72.2 +/-3.3)%, and has an obvious control effect on tobacco black shank.
Compared with the prior art, the invention has the following advantages:
(1) The isoquinoline alkaloid compound is extracted and separated from thalictrum amazonicum for the first time, has an inhibition effect on phytophthora nicotianae which is remarkably superior to that of agricultural streptomycin, and has a control effect on tobacco black shank of (72.2 +/-3.3)%. The application of the compound provides a novel efficient and safe biological pesticide molecular structure for preventing and treating the black-stem disease of tobacco.
(2) The production raw material (Asia-Europe thalictrum) of the invention has wide distribution, large biological output, high alkaloid content, simple compound preparation process, low production cost and easy realization of industrialized application.
Drawings
FIG. 1 is a nuclear magnetic resonance carbon spectrum of an isoquinoline alkaloid compound of the present invention;
FIG. 2 is the NMR spectrum of isoquinoline alkaloid compounds of the present invention;
FIG. 3 is a graph relating the major HMBC groups of isoquinoline alkaloids.
Detailed Description
The present invention is further described in detail with reference to the drawings and examples, which are not intended to limit the technical scope of the present invention, and all changes and equivalents which are made based on the teachings of the present invention should fall within the protective scope of the present invention.
Example 1
Preparation of isoquinoline Alkaloids Compound C 12 H 11 NO 2 The method comprises the steps of extract extraction, silica gel column chromatography and high-pressure liquid chromatography separation, and specifically comprises the following steps:
(1) Extracting the extractum: taking the whole plant of the Asia-Europe thalictrum, drying in the sun, and crushing to 35-60 meshes. Weighing 3.5-5.2 kg of crushed sample, placing the sample in a 20L glass reaction kettle, adding 8-12L of 95% ethanol, performing reflux extraction for 35-60 min, and filtering out an extracting solution; and adding 8-12L of 95% ethanol into the filter residue again, performing reflux extraction for 35-60 min, and filtering an extracting solution. Combining the two extracting solutions, concentrating to a small volume, diluting with 4-6L of 3% tartaric acid solution, and extracting with 4-6L of ethyl acetate for 2 times. After extraction, the water phase is saturated by sodium carbonate, and is extracted for 2 times by 4-6L ethyl acetate, and the extracted ethyl acetate phases are combined and concentrated under reduced pressure to obtain 85.6-128.5 g of alkaloid part extract.
(2) Silica gel column chromatography: dissolving the extract by pure methanol or pure ethanol or pure acetone with the weight ratio of 1.5-3 times, mixing the extract with 120-280 g (80-100 meshes) of crude silica gel, drying, performing column chromatography by 260-440 g of silica gel (150-200 meshes), and performing chloroform: acetone (20.
(3) High-pressure liquid chromatography separation and purification: and (4) selecting an elution part of 6: using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reverse phase column of Agilent company, using 40% methanol water solution as mobile phase, flow rate is 20mL/min, ultraviolet detector detection wavelength is 292nm, collecting 22.7min chromatographic peak, accumulating for multiple times, evaporating to dryness to obtain crude compound. Dissolving the crude product with methanol, purifying with Sephadex column to obtain pure product.
The raw material of the Asian European thaliana used by the invention is not limited by regions and varieties, and can be realized, and the invention is further explained by the raw material of the Asian European thaliana from Yunnan:
example 2
The crabgrass asia sample is from Yunnan university. Drying the whole plant of Asia-Europe Thalictrum, and pulverizing to about 45 mesh. Weighing 3.8kg of crushed sample, placing the sample in a 20L glass reaction kettle, adding 8L of 95% ethanol, performing reflux extraction for 38min, and filtering out an extracting solution; adding 8L of 95% ethanol into the residue again, reflux extracting for 40min, filtering to remove extractive solution, mixing the two extractive solutions, and concentrating to small volume. Then diluted with 6L of 3% tartaric acid solution and extracted 2 times with 6L of ethyl acetate. After extraction, the water phase is saturated with sodium carbonate, extracted for 2 times with 6L ethyl acetate again, the extracted ethyl acetate phases are combined, and the alkaloid part extract 86.4g is obtained by decompression and concentration. Mixing the extract with 120g (80-100 mesh) of crude silica gel, oven drying, performing column chromatography with 280g of silica gel (150-200 mesh), chloroform: acetone (20. The elution fraction of 6: using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reverse phase column of Agilent company, using 40% methanol water solution as mobile phase, flow rate is 20mL/min, ultraviolet detector detection wavelength is 292nm, collecting 22.7min chromatographic peak, accumulating for multiple times, evaporating to dryness to obtain crude compound. Dissolving the crude product with methanol, purifying with Sephadex column to obtain pure compound.
Example 3
The Asian crabgrass sample is from Yunnan Chuxiong. Drying the whole plant of Asia-Europe Thalictrum, and pulverizing to 55 mesh. Weighing 4.6kg of the crushed sample, placing the crushed sample in a 20L glass reaction kettle, adding 12L of 95% ethanol, performing reflux extraction for 25min, and filtering out an extracting solution; adding 95% ethanol 12L into the residue, reflux extracting for 30min, filtering to remove extractive solution, mixing the two extractive solutions, concentrating to small volume, diluting with 3% tartaric acid solution 10L, and extracting with 10L ethyl acetate for 2 times. After extraction, the water phase is saturated with sodium carbonate, and extracted with 10L ethyl acetate for 2 times, and the extracted ethyl acetate phases are combined and concentrated under reduced pressure to obtain 116.8g of alkaloid part extract. Mixing the extract with 200g (80-100 mesh) of crude silica gel, oven drying, and performing column chromatography with 380g of silica gel (150-200 mesh) under the conditions of chloroform: acetone (20. And (4) selecting an elution part of 6: using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reverse phase column of Agilent company, using 40% methanol water solution as mobile phase, flow rate is 20mL/min, ultraviolet detector detection wavelength is 292nm, collecting 22.7min chromatographic peak, accumulating for multiple times, evaporating to dryness to obtain crude compound. Dissolving the crude product with methanol, purifying with Sephadex column to obtain pure compound.
Example 4
The Asia-Europe Thalictrum aquilegifolium sample is from Yunnan Jianchuan. Drying the whole plant of Asia-Europe Thalictrum, and pulverizing to 38 mesh. Weighing 5.0kg of the crushed sample, placing the crushed sample in a 20L glass reaction kettle, adding 12L of 95% ethanol, performing reflux extraction for 30min, and filtering out an extracting solution; adding 10L of 95% ethanol into the filter residue again, reflux extracting for 50min, filtering to remove the extractive solution, mixing the two extractive solutions, concentrating to small volume, diluting with 10L of 3% tartaric acid solution, and extracting with 10L of ethyl acetate for 2 times. After extraction, the water phase is saturated with sodium carbonate, and is extracted with 8L ethyl acetate for 2 times, and the extracted ethyl acetate phases are combined and concentrated under reduced pressure to obtain 124.7g of alkaloid part extract. Mixing the extract with 200g (80-100 mesh) of crude silica gel, oven drying, performing column chromatography with 440g of silica gel (150-200 mesh), chloroform: the acetone (20. And (4) selecting an elution part of 6: using Zorbax PrepHT GF (21.2 mm multiplied by 25 cm) reversed phase column of Agilent company, using 40% methanol water solution as mobile phase, flow rate is 20mL/min, ultraviolet detector detection wavelength is 292nm, collecting 22.7min chromatographic peak, accumulating for multiple times, evaporating to dryness to obtain crude compound. Dissolving the crude product with methanol, purifying with Sephadex column to obtain pure product.
Example 5
Identification of the structure of a compound
The isoquinoline alkaloid compounds prepared in example 2 were collected and measured by the following method. The compound is yellow jelly, and HRESI-MS shows that the peak of the quasi-molecular ion is 224.0680[ M + Na ]] + Is combined with 1 H and 13 c NMR spectrum to determine the molecular formula of C 12 H 11 NO 2 Are alkaloid compounds. The infrared spectrum of the compound showed that the compound had a hydroxyl group (3389 cm) -1 ) Carbonyl group (1668 cm) -1 ) And aromatic rings (1615, 1547 and 1442 cm) -1 ) Signal, ultraviolet spectrumThe presence of aromatic ring structures in the compounds was also confirmed by the absorption maxima at 212, 258, 292 and 334 nm. Process for preparing compounds 1 H、 13 C-NMR and DEPT spectra (Table-1) showed that it contained 12 carbons and 11 hydrogens. The method comprises the following steps: a 1,6, 7-substituted isoquinoline core (C-1 to C-10H-3, H-4, H-5, and H-8), an acetyl group (-CO-CH 3 (ii) a C-1 'and C-2'; h 3 -2 '), one methyl group (C-3'; h 3 -2') and a phenolic hydroxyl group (. Delta.)) H 10.75 s). The isoquinoline parent nucleus can exist through H-3, C-1, C-4 and C-10; h-4 and C-3, C-5, C-9, C-10; h-5 and C-4, C-9, C-10; and HMBC correlation of H-8 and C-1, C-9, C-10 (FIG. 3).
After the parent nucleus of the compound is determined, the positions of the remaining substituents (acetyl, methyl and phenolic hydroxyl) can be determined by further analysis of its HMBC correlation spectrum. By acetylmethyl hydrogen H 3 -2′(δ H 2.46 And C-1 (. Delta.) C 156.5 HMBC correlation of (a) to (b), the substitution of the acetyl group at C-1 can be determined; by aromatic ring methyl hydrogen H 3 -3′(δ H 2.30 And C-6 (. Delta.) C 160.1)、C-7(δ C 154.0 And C-8 (. Delta.) C 127.5 And H-8 (. Delta.) H 8.49 C-3' (delta) C 15.2 HMBC correlation of) can determine that the methyl substitution is at the C-7 position; the substitution of the phenolic hydroxyl group at the C-6 position can be achieved by a phenolic hydroxyl hydrogen (delta) H 10.75 And C-5 (. Delta.) C 112.4)、C-6(δ C 160.1)、 C-7(δ C 132.2 ) was confirmed. The structure of this compound was confirmed and the compound was named: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone.
Example 6
The compound prepared in example 3 was taken as a yellow gum. The determination method was the same as in example 5, and it was confirmed that the compound prepared in example 3 was 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone, which is the above-mentioned isoquinoline alkaloid compound.
Example 7
The compound prepared in example 4 was taken as a yellow gum. The compound prepared in example 4 was confirmed to be the above-mentioned 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone in the same manner as in example 5.
Example 8
Any of the isoquinoline alkaloid compounds prepared in examples 1-4 are tested for activity against tobacco black shank as follows:
(1) Adding 1000mL of water into oatmeal, heating the oatmeal in a boiling water bath for 1 hour, filtering the oatmeal by using gauze, adding the water for supplementing 1000mL, then adding sugar and agar, heating the oatmeal to completely melt the agar, filtering the oatmeal in a triangular flask by using the gauze (absorbent cotton is added in the middle) while the oatmeal is hot, sterilizing the oatmeal for 20min at 121 ℃ and 15 pounds, taking out the oatmeal and cooling the oatmeal to about 45 ℃, adding ampicillin (5 mg/100 mL) into an aseptic operation table, uniformly mixing the ampicillin and the ampicillin, pouring the ampicillin into the flat dish, culturing the ampicillin for 48 hours at 28 ℃, and checking the ampicillin for later use after the ampicillin is checked to be aseptic.
(2) Circular filter paper with diameter of 5mm is placed in a culture dish, sterilized under high pressure for 30min under 15 pounds, and then is respectively immersed in 20 mu M compound, 75% ethanol solution and sterilized distilled water after being dried. Respectively sucking 0.2mL of fresh bacterial liquid of phytophthora nicotianae on oatmeal agar medium plates by using a sterile suction pipe on a sterile operating table. Coating with triangular glass coating rod, lightly sticking the filter paper sheets onto corresponding flat plates with tweezers, culturing at 28 deg.C, observing experimental results, and measuring the size of antibacterial zone. Meanwhile, agricultural streptomycin is used as a positive control.
(3) The test result shows that: the diameter of the inhibition zone of the isoquinoline alkaloid compounds is 14.6 +/-1.2 mm, and the diameter of the inhibition zone of the positive control agricultural streptomycin is 12.2 +/-1.0 mm. The compound has the advantages of obviously better effect of inhibiting phytophthora nicotianae than positive control agricultural streptomycin and outstanding activity of inhibiting black shank.
(4) Transplanting tobacco seedlings into flowerpots with the diameter of 10cm and the height of 10cm, wherein the culture medium is as follows: sterilized soil, peat and perlite culture (2. After transplanting the slow seedlings, 10g of bacterial grains are added to roots, the tobacco seedlings are placed in a climatic chamber for cultivation, the temperature is 30 ℃ in the daytime, the temperature is 28 ℃ in the dark at night, the ratio of light to dark (12h. To make the tobacco seedling attack. Before the black shank disease occurs, the tobacco seedlings are subjected to root irrigation treatment by using 20 mu M of the compound, and 10mL of the compound is irrigated to each tobacco seedling; the pouring is carried out for 2 times. Each of 10 plants was treated 3 times and 14 days later, the disease onset was investigated, and the disease index was calculated. The results show that: the compound has the effect of controlling the tobacco black shank of (72.2 +/-3.3)%, and has remarkable effect of controlling the tobacco black shank.
Claims (4)
1. An isoquinoline alkaloid compound with molecular formula of C 12 H 11 NO 2 Having the following structural formula:
the compound was named: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone, english name: 1- (6-hydroxy-7-methylisoquinolin-1-yl) ethanone.
2. A process for the preparation of isoquinoline alkaloid compounds as claimed in claim 1 comprising the steps of:
(1) Extracting the extractum: drying the whole grass of thalictrum foeniculaceum, crushing to 35-60 meshes, placing the crushed sample in a glass reaction kettle, performing reflux extraction for 2 times by using 95% ethanol, combining the two extracting solutions, concentrating to a small volume, diluting by using a 3% tartaric acid solution, and extracting for 2 times by using ethyl acetate; saturating the water phase with sodium carbonate, extracting with ethyl acetate for 2 times, mixing the extracted ethyl acetate phases, and concentrating under reduced pressure to obtain alkaloid extract;
(2) Silica gel column chromatography: dissolving the extract obtained in the step (1) by using 1.5-3 times of pure methanol or pure ethanol or pure acetone, mixing the extract with 0.8-2.5 times of 80-100 meshes of silica gel, and drying; silica gel filled in the column is 150-200 meshes, and the dosage of the silica gel is 3-6 times of the weight of the extract; gradient elution is performed with chloroform-acetone solutions of 20;
(3) High-pressure liquid chromatography separation and purification: and (3) taking 6 parts of the column chromatography gradient eluent for high-pressure liquid chromatography separation and purification: using Zorbax PrepHT GF of Agilent company, a 21.2mm multiplied by 25cm reverse phase column, using 40% methanol water solution as a mobile phase, the flow rate is 20mL/min, the detection wavelength of an ultraviolet detector is 292nm, collecting a chromatographic peak of 22.7min, accumulating for multiple times, and evaporating to dryness to obtain the isoquinoline alkaloid compound.
3. The method of claim 2, wherein: and (3) dissolving the compound obtained after the separation and purification by the high pressure liquid chromatography in pure methanol, and separating by a sephadex column chromatography with the pure methanol as a mobile phase to obtain a yellow jelly, namely a pure compound.
4. The use of isoquinoline alkaloid compounds as claimed in claim 1 in the control of tobacco black shank.
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