CN102190698B - Alga endophytic fungi diterpenoid alkaloid compound, preparation method thereof and application thereof - Google Patents
Alga endophytic fungi diterpenoid alkaloid compound, preparation method thereof and application thereof Download PDFInfo
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- -1 diterpenoid alkaloid compound Chemical class 0.000 title claims abstract description 30
- 241000233866 Fungi Species 0.000 title claims abstract description 29
- 229930013930 alkaloid Natural products 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 238000004809 thin layer chromatography Methods 0.000 claims abstract description 26
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 240000006439 Aspergillus oryzae Species 0.000 claims abstract description 10
- 239000000287 crude extract Substances 0.000 claims abstract description 10
- 238000010828 elution Methods 0.000 claims abstract description 10
- 239000003960 organic solvent Substances 0.000 claims abstract description 10
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 7
- 238000000855 fermentation Methods 0.000 claims abstract description 6
- 230000004151 fermentation Effects 0.000 claims abstract description 6
- 230000000749 insecticidal effect Effects 0.000 claims abstract description 6
- 238000000926 separation method Methods 0.000 claims abstract description 4
- 238000000746 purification Methods 0.000 claims abstract description 3
- 239000007788 liquid Substances 0.000 claims description 20
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 claims description 14
- 239000003208 petroleum Substances 0.000 claims description 14
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Substances CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 12
- 238000010898 silica gel chromatography Methods 0.000 claims description 12
- 239000000463 material Substances 0.000 claims description 9
- 238000003810 ethyl acetate extraction Methods 0.000 claims description 8
- 238000001641 gel filtration chromatography Methods 0.000 claims description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 235000001018 Hibiscus sabdariffa Nutrition 0.000 claims description 5
- 235000005291 Rumex acetosa Nutrition 0.000 claims description 5
- 240000007001 Rumex acetosella Species 0.000 claims description 5
- 235000003513 sheep sorrel Nutrition 0.000 claims description 5
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000003905 agrochemical Substances 0.000 claims description 4
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical compound OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 claims description 4
- 238000000638 solvent extraction Methods 0.000 claims description 4
- 230000003068 static effect Effects 0.000 claims description 4
- UXTMROKLAAOEQO-UHFFFAOYSA-N chloroform;ethanol Chemical compound CCO.ClC(Cl)Cl UXTMROKLAAOEQO-UHFFFAOYSA-N 0.000 claims description 3
- 239000002917 insecticide Substances 0.000 claims description 2
- 239000000575 pesticide Substances 0.000 abstract description 7
- 150000001875 compounds Chemical class 0.000 abstract description 6
- 241000238582 Artemia Species 0.000 abstract description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 abstract description 4
- 238000004440 column chromatography Methods 0.000 abstract 3
- 239000003480 eluent Substances 0.000 abstract 3
- BPQQTUXANYXVAA-UHFFFAOYSA-N Orthosilicate Chemical compound [O-][Si]([O-])([O-])[O-] BPQQTUXANYXVAA-UHFFFAOYSA-N 0.000 abstract 1
- 239000000377 silicon dioxide Substances 0.000 abstract 1
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 10
- 229910052736 halogen Inorganic materials 0.000 description 9
- 150000002367 halogens Chemical class 0.000 description 9
- 229930002995 diterpene alkaloid Natural products 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000238426 Anostraca Species 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000595940 Notostraca Species 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 208000031513 cyst Diseases 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 230000012447 hatching Effects 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 231100000225 lethality Toxicity 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 229960001866 silicon dioxide Drugs 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000006273 synthetic pesticide Substances 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 238000005160 1H NMR spectroscopy Methods 0.000 description 1
- 241000238421 Arthropoda Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 238000003811 acetone extraction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000005276 aerator Methods 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 238000010170 biological method Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 210000001136 chorion Anatomy 0.000 description 1
- 239000012230 colorless oil Substances 0.000 description 1
- AAOVKJBEBIDNHE-UHFFFAOYSA-N diazepam Chemical compound N=1CC(=O)N(C)C2=CC=C(Cl)C=C2C=1C1=CC=CC=C1 AAOVKJBEBIDNHE-UHFFFAOYSA-N 0.000 description 1
- 150000003800 diterpene alkaloid derivatives Chemical class 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000003808 methanol extraction Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 239000002352 surface water Substances 0.000 description 1
Abstract
The invention relates to a pesticide field, and concretely relates to an alga endophytic fungi diterpenoid alkaloid compound, a preparation method thereof and an application thereof. The preparation method comprises the following steps: Aspergillus oryzae cf-2 is inoculated into a medium and ferments statically, a fermentation broth is extracted and condensed through ethyl acetate to obtain a first condensate, mycelium is extracted with an organic solvent and is extracted and condensed with ethyl acetate to obtain a second condensate which is combined with the first condensate to obtain a crude extract; the crude extract is treated with silica column chromatography and is treated with gradient elution by an organic solvent, and an eluent is collected and is detected through thin layer chromatography (TLC); and an eluent component obtained through gradient elution with an eluent volume ratio of 10 to 15:1 is collected, and is treated with gel column chromatography, silicate column chromatography and the TLC for separation and purification to obtain the target compound. The alga endophytic fungi diterpenoid alkaloid compound with a concentration of 100 <mu>g/ml has a fatality of 61.9% to Artemia species according to insecticidal activity tests.
Description
Technical field
The present invention relates to field of pesticides, specifically a kind of alga endophytic fungi diterpenoid alkaloid compound and preparation thereof and application.
Background technology
Along with the chemical synthetic pesticide passing of duration of service, the detrimentally affect of its generation is more and more outstanding.Chemical pesticide is produced on a large scale and is unrestrictedly used in a large number, 70%～80% agricultural chemicals directly is penetrated in the environment, residual very serious, soil, surface water, underground water are polluted, the stepping of going forward side by side enters biologic chain, and biological and human health are all caused direct and potential harm.Behind China's accession to the WTO, " green barrier " that causes because chemical pesticide is residual seriously restricting the outlet of agricultural products in China, and the traditional agriculture industry is faced with larger risk and challenge.The Ministry of Agriculture has implemented " pollution-free food action plan " in China from calendar year 2001, be the key measure of implementing this target and use public nuisance-free agricultural chemicals, so people invest the biological pesticide more friendly to environment to sight.Compare with traditional chemical synthetic pesticide, biological pesticide has people and animals and non-target organism safety, and environment compatibility is good, is difficult for producing the advantages such as resistance.The Application and Development of biological pesticide is to human health, and the Sustainable development of environment protection and agricultural all is extremely important.
Summary of the invention
The purpose of this invention is to provide a kind of alga endophytic fungi diterpenoid alkaloid compound and preparation thereof and application.
For achieving the above object, the technical solution used in the present invention is:
A kind of alga endophytic fungi diterpenoid alkaloid compound: described alga endophytic fungi diterpenoid alkaloid compound is suc as formula shown in (I).
Preparation method's following steps:
1) aspergillus oryzae (Aspergillus oryzae) cf-2 was inoculated in the fungi liquid substratum static fermentation 20-25 days, filter, fermented liquid is concentrated through ethyl acetate extraction, the mycelium organic solvent extraction of collecting, then concentrated through ethyl acetate extraction again, fermented liquid gained enriched material and mycelium gained enriched material merge, and are crude extract;
Described aspergillus oryzae (Asp.oryzae) cf-2, it is stored in Chinese Typical Representative culture collection center C CTCC on March 1st, 2010, and deposit number is CCTCC M 2010045, and strain number is cf-2;
2) get step 1) in crude extract carry out silica gel column chromatography, carry out gradient elution with organic solvent, collect elutriant, elutriant detects through thin-layer chromatography;
3) collect step 2) in effluent volume than 10-15: the elution fraction of 1 gradient, the elution fraction of collecting is carried out gel filtration chromatography, silica gel column chromatography and thin-layer chromatography separation and purification, it is the 0.3-0.4 component that purifying is collected the Rf value, namely gets suc as formula the alga endophytic fungi diterpenoid alkaloid compound shown in (I).
Step 1) the organic solvent extraction liquid in is in chloroform, acetone, ethyl acetate, ethanol or the methyl alcohol one or more.Step 2) organic solvent in is petroleum ether-ethyl acetate or sherwood oil-acetone.Step 3) described gel filtration chromatography elutriant is volume ratio 1-2: 1 chloroform-methanol or chloroform-ethanol; The silica gel column chromatography elutriant is volume ratio 5-15: 1 than petroleum ether-ethyl acetate or sherwood oil-acetone; The thin-layer chromatography developping agent is that volume ratio is 5: 1 petroleum ether-ethyl acetate.Step 3) collects the component that detects the sorrel spot through TLC during described gel filtration chromatography; Collect during silica gel column chromatography with effluent volume than 10-15: the elutriant of 1 gradient.
Use: described alga endophytic fungi diterpenoid alkaloid compound has insecticidal action.Described alga endophytic fungi diterpenoid alkaloid compound can be used as agricultural chemical insecticide.
The present invention has the following advantages:
The alga endophytic fungi diterpenoid alkaloid compound that the present invention obtains through extraction, separation by fungi aspergillus oryzae (Asp.oryzae) the cf-2 fermentation that is located away from the different pipe algae of marine red alga, the lethality rate to the halogen worm when the insecticidal activity experiment draws compound in 100 mcg/ml is 61.9%.
Embodiment:
Below in conjunction with embodiment the present invention is further elaborated.
Embodiment 1
Alga endophytic fungi diterpenoid alkaloid compound is suc as formula shown in (I).
Embodiment 2
Preparation method suc as formula the alga endophytic fungi diterpenoid alkaloid compound shown in (I):
Well-grown fungi aspergillus oryzae (Asp.oryzae) cf-2 bacterial classification on the plate of making even, it is stored in Chinese Typical Representative culture collection center C CTCC on March 1st, 2010, and deposit number is CCTCC M2010045, and strain number is cf-2; Being cut into small pieces is inoculated in the PDB liquid nutrient medium, puts the 300mL substratum in every 1L triangular flask, and totally 30 bottles, the static fermentation of room temperature 23 days, the ethyl acetate killing fungus of adding fermented liquid 1/2nd volumes filters, and collects respectively mycelium and fermented liquid.Described PDB liquid nutrient medium consists of every liter and contains 200 milliliters of murphy juices, glucose 20 grams, peptone 5 grams, yeast extract paste 3 grams, 500 milliliters of Chen Haishui, 300 milliliters of distilled water.
Collect approximately 9L of fermented liquid, use ethyl acetate extraction three times, concentrating under reduced pressure; Use 1: 1 chloroform-methanol extraction of volume ratio three times after mycelium is pulverized, use again ethyl acetate extraction, concentrating under reduced pressure; It is similar that enriched material detects its result through thin-layer chromatography, and merging fermented liquid and the two-part enriched material of mycelium is crude extract 8g.Crude extract is carried out 200-300 purpose silica gel column chromatography, with petroleum ether-ethyl acetate with 100: 0,50: 1,20: 1,10: 1,5: 1,1: 1 to 0: 100 gradient is carried out wash-out, collects respectively elutriant, uses thin-layer chromatography (TLC) to detect (using aubepine-sulfuric acid when thin-layer chromatography detects as developer) again, judge, merge identical or similar portions according to the Rf value, obtain 22 components (1-22).Component 15 is namely carried out gel column again with the component under 10: 1 gradient elutions of petroleum ether-ethyl acetate, silicagel column separates with thin-layer chromatography, elutriant 1: 1 chloroform-methanol of volume ratio during gel filtration chromatography, component is developping agent through TLC detection employing petroleum ether-ethyl acetate under the wash-out, collect the component of sorrel spot, carry out wash-out with gradient at 10: 1 to 5: 1 petroleum ether-ethyl acetate during silica gel column chromatography, collect the component under 10: 1 the petroleum ether-ethyl acetate wash-out, the petroleum ether-ethyl acetate that separates with 5: 1 through thin-layer chromatography is developping agent, collecting the Rf value is the component of 0.3-0.4, get compound (3.4 milligrams) shown in the formula (I), detect through TLC, be single, evenly the sorrel spot is defined as pure compound.Through Spectrum Analysis, its Structural Identification is a kind of new diterpene alkaloid, structural formula shown in (I), called after Asporyzin B.
This compound has following physics and chemistry and spectral characteristic:
Colorless oil, proton nmr spectra (
1H NMR, CDCl
3, 500MHz) δ 7.15 (br s), 1.62 (m), 1.78 (m), 1.59 (m), 1.88 (m), 3.17 (dd, 12.2,3.2), 4.77 (m), 1.14 (m), 1.87 (m), (1.50 dd, 13.0,2.6), 1.34 (m), 1.43 (m), 1.44 (m), 1.57 (m), 2.84 (m), (2.38 dd, 18.0,4.9), 2.99 (dd, 18.0,3.4), 7.67 (dd, 7.6,1.5), 7.37 (t, 7.4), (7.48 td, 7.6,1.5), (7.08 d, 7.8), 1.37 (s), 1.01 (s), 5.31 (d, 9.0), 1.66 (s), 1.72 (s), 0.79 (s); Carbon-13 nmr spectra (
13C NMR, CDCl
3, 125MHz) δ 176.6 (s), 59.7 (s), 42.1 (s), 33.9 (t), 22.3 (t), 85.5 (d), 73.8 (d), 48.9 (t), 45.0 (s), 45.0 (d), 23.9 (t), 30.0 (t), 34.9 (d), 48.7 (t), 202.9 (s), 134.8 (s), 129.3 (d), 127.6 (d), 132.7 (d), 127.2 (d), 136.9 (s), 15.3 (q), 19.6 (q), 126.7 (d), 134.9 (s), 18.0 (q), 25.9 (q), 15.5 (q); High resolution mass spectrum (HREIMS) [M]
+M/z 435.2769, calculated value 435.2773.
Embodiment 3
The insecticidal activity experiment:
The halogen worm (Brine Shrimp) also claim the salt solution fairy shrimp, belongs to Arthropoda in the classification, Crustachia, and Anostraca, salt solution fairy shrimp section, genus artemia, the halogen worm attracts widespread attention as a kind of important and good laboratory animal material always.Adopt this insect that cultivate in the laboratory under the control condition, borrow it to come the power of assessing compound insecticidal activity.
The hatching of artemia cysts: get 100 milligrams of artemia cysts and place 500 ml beakers, add 400 milliliters of artificial seawaters, slowly inflate with a little aerator pump, chorion and unhatched ovum are removed in room temperature hatching 24 hours, and halogen worm larva continues to cultivate 24 hours, and is for subsequent use.
The biological method that causes death of halogen worm: according to the improved method of Solis, get 96 porocyte culture plates, every hole adds the artificial seawater liquid that 190 microlitres contain 10 left and right sides halogen worm larvas, makes the test cultures plate.The sample sets of blank group and each concentration is respectively established three parallel holes, the blank group adds 10 microlitre dimethyl sulfoxide solvents (DMSO), sample sets adds 10 microlitres, concentration is the DMSO solution of the diterpene alkaloid compounds shown in the 2.0 mg/ml formulas (I), the DMSO solution of the diterpene alkaloid compounds shown in its Chinese style (I) is take DMSO as solvent, and the diterpene alkaloid compounds shown in the formula (I) is solute.After the incubated at room temperature 24 hours, under the binocular anatomical lens, detect the dead individual amount of counting halogen worm.
The halogen worm is biological to cause death and actively represents with corrected mortality, and calculation formula is as follows:
Corrected mortality=(control group survival rate-treatment group survival rate)/control group survival rate * 100%
Experimental result: the diterpene alkaloid compounds of the acquisition in above-described embodiment lethality rate to the halogen worm when 100 mcg/ml is 61.9%.
Embodiment 4
Difference from Example 2 is
Well-grown fungi aspergillus oryzae (Asp.oryzae) cf-2 bacterial classification on the plate of making even, be cut into small pieces and be inoculated in the PDB liquid nutrient medium, put the 300mL substratum in every 1L triangular flask, totally 30 bottles, the static fermentation of room temperature 23 days, add 1/2nd volume ethyl acetate killing fungus, filter, collect respectively mycelium and fermented liquid.Described PDB liquid nutrient medium consists of every liter and contains 200 milliliters of murphy juices, glucose 20 grams, peptone 5 grams, yeast extract paste 3 grams, 500 milliliters of Chen Haishui, 300 milliliters of distilled water.
Collect approximately 9L of fermented liquid, use ethyl acetate extraction three times, concentrating under reduced pressure; Use acetone extraction three times after mycelium is pulverized, use again ethyl acetate extraction, concentrating under reduced pressure; It is similar that enriched material detects its result through thin-layer chromatography, and merging fermented liquid and the two-part enriched material of mycelium is crude extract 8g.Crude extract is carried out 200-300 purpose silica gel column chromatography, with sherwood oil-acetone with 100: 0,50: 1,20: 1,15: 1,10: 1,5: 1,1: 1 to 0: 100 gradient was carried out wash-out, collect respectively elutriant, use again thin-layer chromatography (TLC) to detect (using aubepine-sulfuric acid when thin-layer chromatography detects as developer), judge, merge identical or similar portions according to the Rf value, obtain 22 components (1-22).The component of component 15 under namely take sherwood oil-acetone volume ratio as 15: 1 gradient elutions carried out gel column again, silicagel column separates with thin-layer chromatography, elutriant is chloroform-ethanol of 2: 1 of volume ratio during gel filtration chromatography, component is developping agent through TLC detection employing petroleum ether-ethyl acetate under the wash-out, collect the component of sorrel spot, carry out wash-out with gradient at sherwood oil-acetone of 15: 1 to 10: 1 during silica gel column chromatography, collect the component under 15: 1 the sherwood oil-acetone wash-out, the petroleum ether-ethyl acetate that separates with 5: 1 through thin-layer chromatography is developping agent, collecting the Rf value is the component of 0.3-0.4, gets the diterpene alkaloid compounds shown in the formula (I).
Claims (7)
1. alga endophytic fungi diterpenoid alkaloid compound, it is characterized in that: described alga endophytic fungi diterpenoid alkaloid compound is suc as formula shown in (I)
2. preparation method by alga endophytic fungi diterpenoid alkaloid compound claimed in claim 1 is characterized in that the following steps preparation:
1) aspergillus oryzae (Aspergillus oryzae) cf-2 was inoculated in the fungi liquid substratum static fermentation 20-25 days, filter, fermented liquid is concentrated through ethyl acetate extraction, the mycelium organic solvent extraction of collecting, then concentrated through ethyl acetate extraction again, fermented liquid gained enriched material and mycelium gained enriched material merge, and are crude extract;
Described aspergillus oryzae (Asp.oryzae) cf-2, it is stored in Chinese Typical Representative culture collection center C CTCC on March 1st, 2010, and deposit number is CCTCC M 2010045, and strain number is cf-2;
2) get step 1) in crude extract carry out silica gel column chromatography, carry out gradient elution with organic solvent, collect elutriant, elutriant detects through thin-layer chromatography, described organic solvent is petroleum ether-ethyl acetate or sherwood oil-acetone;
3) collect step 2) in effluent volume than 10-15: the elution fraction of 1 gradient, the elution fraction of collecting is carried out gel filtration chromatography, silica gel column chromatography and thin-layer chromatography separation and purification, it is the 0.3-0.4 component that purifying is collected the Rf value, namely get suc as formula the alga endophytic fungi diterpenoid alkaloid compound shown in (I), described thin-layer chromatography developping agent is that volume ratio is 5: 1 petroleum ether-ethyl acetate.
3. by the preparation method of alga endophytic fungi diterpenoid alkaloid compound claimed in claim 2, it is characterized in that: step 1) in organic solvent extraction liquid be in chloroform, acetone, ethyl acetate, ethanol or the methyl alcohol one or more.
4. by the preparation method of alga endophytic fungi diterpenoid alkaloid compound claimed in claim 2, it is characterized in that: step 3) described gel filtration chromatography elutriant is volume ratio 1-2: 1 chloroform-methanol or chloroform-ethanol; The silica gel column chromatography elutriant is volume ratio 5-15: 1 petroleum ether-ethyl acetate or sherwood oil-acetone.
5. by the preparation method of alga endophytic fungi diterpenoid alkaloid compound claimed in claim 2, it is characterized in that: step 3) collect the component that detects the sorrel spot through TLC during described gel filtration chromatography; Collect during silica gel column chromatography with effluent volume than 10-15: the elutriant of 1 gradient.
6. application by alga endophytic fungi diterpenoid alkaloid compound claimed in claim 1, it is characterized in that: described alga endophytic fungi diterpenoid alkaloid compound has insecticidal action.
7. by the application of alga endophytic fungi diterpenoid alkaloid compound claimed in claim 6, it is characterized in that: described alga endophytic fungi diterpenoid alkaloid compound can be used as agricultural chemical insecticide.
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| CN101225078A (en) * | 2008-01-18 | 2008-07-23 | 中国科学院海洋研究所 | Sea weed sesquiterpenoids as well as preparation method and application thereof |
| CN101230068A (en) * | 2008-01-25 | 2008-07-30 | 中国科学院海洋研究所 | Polyether triterpene compound as well as preparation method and application thereof |
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| CN101225078A (en) * | 2008-01-18 | 2008-07-23 | 中国科学院海洋研究所 | Sea weed sesquiterpenoids as well as preparation method and application thereof |
| CN101230068A (en) * | 2008-01-25 | 2008-07-30 | 中国科学院海洋研究所 | Polyether triterpene compound as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| Nai-Yun Ji et al..Highly Oxygenated Triterpenoids from the Marine Red Alga Laurencia mariannensis (Rhodomelaceae).《Helvetica Chimica Acta》.2008,第91卷(第10期),1940-1946. * |
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