CN115444003B - Method for preparing high-drug-loading phoxim nanostructure lipid carrier based on microfluidics - Google Patents
Method for preparing high-drug-loading phoxim nanostructure lipid carrier based on microfluidics Download PDFInfo
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- CN115444003B CN115444003B CN202211039810.9A CN202211039810A CN115444003B CN 115444003 B CN115444003 B CN 115444003B CN 202211039810 A CN202211039810 A CN 202211039810A CN 115444003 B CN115444003 B CN 115444003B
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- Prior art keywords
- phoxim
- lipid
- lipid carrier
- drug
- nanostructure
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- 229950001664 phoxim Drugs 0.000 title claims abstract description 161
- ATROHALUCMTWTB-OWBHPGMISA-N phoxim Chemical compound CCOP(=S)(OCC)O\N=C(\C#N)C1=CC=CC=C1 ATROHALUCMTWTB-OWBHPGMISA-N 0.000 title claims abstract description 161
- 150000002632 lipids Chemical class 0.000 title claims abstract description 125
- 239000002086 nanomaterial Substances 0.000 title claims abstract description 48
- 238000000034 method Methods 0.000 title claims abstract description 35
- 238000011068 loading method Methods 0.000 title abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000007788 liquid Substances 0.000 claims abstract description 22
- 239000002736 nonionic surfactant Substances 0.000 claims abstract description 19
- 239000003945 anionic surfactant Substances 0.000 claims abstract description 16
- 239000007787 solid Substances 0.000 claims abstract description 15
- 239000004064 cosurfactant Substances 0.000 claims abstract description 14
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 22
- WRIDQFICGBMAFQ-UHFFFAOYSA-N (E)-8-Octadecenoic acid Natural products CCCCCCCCCC=CCCCCCCC(O)=O WRIDQFICGBMAFQ-UHFFFAOYSA-N 0.000 claims description 21
- LQJBNNIYVWPHFW-UHFFFAOYSA-N 20:1omega9c fatty acid Natural products CCCCCCCCCCC=CCCCCCCCC(O)=O LQJBNNIYVWPHFW-UHFFFAOYSA-N 0.000 claims description 21
- QSBYPNXLFMSGKH-UHFFFAOYSA-N 9-Heptadecensaeure Natural products CCCCCCCC=CCCCCCCCC(O)=O QSBYPNXLFMSGKH-UHFFFAOYSA-N 0.000 claims description 21
- 239000005642 Oleic acid Substances 0.000 claims description 21
- ZQPPMHVWECSIRJ-UHFFFAOYSA-N Oleic acid Natural products CCCCCCCCC=CCCCCCCCC(O)=O ZQPPMHVWECSIRJ-UHFFFAOYSA-N 0.000 claims description 21
- QXJSBBXBKPUZAA-UHFFFAOYSA-N isooleic acid Natural products CCCCCCCC=CCCCCCCCCC(O)=O QXJSBBXBKPUZAA-UHFFFAOYSA-N 0.000 claims description 21
- ZQPPMHVWECSIRJ-KTKRTIGZSA-N oleic acid Chemical compound CCCCCCCC\C=C/CCCCCCCC(O)=O ZQPPMHVWECSIRJ-KTKRTIGZSA-N 0.000 claims description 21
- 239000001397 quillaja saponaria molina bark Substances 0.000 claims description 21
- 229930182490 saponin Natural products 0.000 claims description 21
- LDVVTQMJQSCDMK-UHFFFAOYSA-N 1,3-dihydroxypropan-2-yl formate Chemical compound OCC(CO)OC=O LDVVTQMJQSCDMK-UHFFFAOYSA-N 0.000 claims description 20
- 150000007949 saponins Chemical class 0.000 claims description 20
- 241001122767 Theaceae Species 0.000 claims description 18
- 238000000520 microinjection Methods 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 8
- BWPGKXYWPBQBPV-MWQJAWBESA-N Theasaponin Chemical compound O([C@H]1[C@H](O)[C@H](O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@@H](O)CO1)O[C@H]1[C@@H]([C@@H](O)[C@H](O)CO1)O)O[C@H]1CC[C@]2(C)[C@H]3CC=C4[C@@]([C@@]3(CC[C@H]2[C@@]1(CO)C)C)(C)C[C@@H](O)[C@@]1(CO)[C@@H](OC(C)=O)[C@@H](C(C[C@H]14)(C)C)OC(=O)C(\C)=C/C)C(O)=O)[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BWPGKXYWPBQBPV-MWQJAWBESA-N 0.000 claims description 6
- BWPGKXYWPBQBPV-ZOADXXHESA-N Theasaponin Natural products O=C(O[C@@H]1[C@@H](OC(=O)C)[C@]2(CO)[C@@H](O)C[C@@]3(C)[C@@]4(C)[C@@H]([C@]5(C)[C@H]([C@@](CO)(C)[C@@H](O[C@@H]6[C@@H](O[C@@H]7[C@H](O[C@@H]8[C@@H](O)[C@H](O)[C@H](O)CO8)[C@H](O)[C@@H](O)CO7)[C@@H](O[C@H]7[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O7)[C@H](O)[C@@H](C(=O)O)O6)CC5)CC4)CC=C3[C@@H]2CC1(C)C)/C(=C/C)/C BWPGKXYWPBQBPV-ZOADXXHESA-N 0.000 claims description 6
- QXNVGIXVLWOKEQ-UHFFFAOYSA-N Disodium Chemical class [Na][Na] QXNVGIXVLWOKEQ-UHFFFAOYSA-N 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 239000005457 ice water Substances 0.000 claims description 5
- 239000012528 membrane Substances 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- ULUAUXLGCMPNKK-UHFFFAOYSA-N Sulfobutanedioic acid Chemical compound OC(=O)CC(C(O)=O)S(O)(=O)=O ULUAUXLGCMPNKK-UHFFFAOYSA-N 0.000 claims description 3
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 229940051841 polyoxyethylene ether Drugs 0.000 claims description 2
- 229920000056 polyoxyethylene ether Polymers 0.000 claims description 2
- SFNALCNOMXIBKG-UHFFFAOYSA-N ethylene glycol monododecyl ether Chemical compound CCCCCCCCCCCCOCCO SFNALCNOMXIBKG-UHFFFAOYSA-N 0.000 claims 1
- 239000012530 fluid Substances 0.000 claims 1
- 239000011148 porous material Substances 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 36
- 238000002360 preparation method Methods 0.000 abstract description 32
- 229940079593 drug Drugs 0.000 abstract description 28
- 238000003860 storage Methods 0.000 abstract description 20
- 241000985245 Spodoptera litura Species 0.000 abstract description 12
- 230000000694 effects Effects 0.000 abstract description 7
- 241000607479 Yersinia pestis Species 0.000 abstract description 6
- 241000255777 Lepidoptera Species 0.000 abstract description 3
- 230000007613 environmental effect Effects 0.000 abstract description 3
- 230000002349 favourable effect Effects 0.000 abstract 1
- 239000000575 pesticide Substances 0.000 description 44
- 239000012071 phase Substances 0.000 description 44
- 238000005538 encapsulation Methods 0.000 description 28
- 239000000203 mixture Substances 0.000 description 24
- 239000003921 oil Substances 0.000 description 22
- 238000009472 formulation Methods 0.000 description 20
- 239000004094 surface-active agent Substances 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 238000012512 characterization method Methods 0.000 description 11
- 239000002245 particle Substances 0.000 description 11
- 239000002552 dosage form Substances 0.000 description 10
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 10
- 239000000523 sample Substances 0.000 description 10
- 238000005338 heat storage Methods 0.000 description 9
- 230000008569 process Effects 0.000 description 8
- 230000014759 maintenance of location Effects 0.000 description 7
- 239000008186 active pharmaceutical agent Substances 0.000 description 6
- 229940088679 drug related substance Drugs 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 230000006872 improvement Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 5
- 239000002994 raw material Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 230000001988 toxicity Effects 0.000 description 5
- 231100000419 toxicity Toxicity 0.000 description 5
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 4
- 241000196324 Embryophyta Species 0.000 description 4
- 240000000249 Morus alba Species 0.000 description 4
- 235000008708 Morus alba Nutrition 0.000 description 4
- 238000002441 X-ray diffraction Methods 0.000 description 4
- 230000002051 biphasic effect Effects 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 229910052802 copper Inorganic materials 0.000 description 4
- 239000010949 copper Substances 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 4
- 238000004945 emulsification Methods 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 238000001069 Raman spectroscopy Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000002329 infrared spectrum Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- VBICKXHEKHSIBG-UHFFFAOYSA-N 1-monostearoylglycerol Chemical compound CCCCCCCCCCCCCCCCCC(=O)OCC(O)CO VBICKXHEKHSIBG-UHFFFAOYSA-N 0.000 description 2
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- 238000001237 Raman spectrum Methods 0.000 description 2
- 239000008346 aqueous phase Substances 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 239000004495 emulsifiable concentrate Substances 0.000 description 2
- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 2
- 230000000749 insecticidal effect Effects 0.000 description 2
- ZXEKIIBDNHEJCQ-UHFFFAOYSA-N isobutanol Chemical compound CC(C)CO ZXEKIIBDNHEJCQ-UHFFFAOYSA-N 0.000 description 2
- -1 lauramidopropyl Chemical group 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000005507 spraying Methods 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- MJJJJEUEZVGFLW-UHFFFAOYSA-N 2-dodecyl-2-sulfobutanedioic acid Chemical compound CCCCCCCCCCCCC(S(O)(=O)=O)(C(O)=O)CC(O)=O MJJJJEUEZVGFLW-UHFFFAOYSA-N 0.000 description 1
- HIQIXEFWDLTDED-UHFFFAOYSA-N 4-hydroxy-1-piperidin-4-ylpyrrolidin-2-one Chemical compound O=C1CC(O)CN1C1CCNCC1 HIQIXEFWDLTDED-UHFFFAOYSA-N 0.000 description 1
- 102000012440 Acetylcholinesterase Human genes 0.000 description 1
- 108010022752 Acetylcholinesterase Proteins 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000251511 Holothuroidea Species 0.000 description 1
- 235000021314 Palmitic acid Nutrition 0.000 description 1
- 235000021355 Stearic acid Nutrition 0.000 description 1
- 231100000605 Toxicity Class Toxicity 0.000 description 1
- 241000819233 Tribulus <sea snail> Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 229940022698 acetylcholinesterase Drugs 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000013871 bee wax Nutrition 0.000 description 1
- 239000012166 beeswax Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 238000003889 chemical engineering Methods 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- MHKKFFHWMKEBDW-UHFFFAOYSA-N dimethyl 2,5-dioxocyclohexane-1,4-dicarboxylate Chemical compound COC(=O)C1CC(=O)C(C(=O)OC)CC1=O MHKKFFHWMKEBDW-UHFFFAOYSA-N 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000021050 feed intake Nutrition 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229930182494 ginsenoside Natural products 0.000 description 1
- 229940089161 ginsenoside Drugs 0.000 description 1
- 125000005456 glyceride group Chemical group 0.000 description 1
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 1
- 229940075507 glyceryl monostearate Drugs 0.000 description 1
- 239000000383 hazardous chemical Substances 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- XUGNVMKQXJXZCD-UHFFFAOYSA-N isopropyl palmitate Chemical compound CCCCCCCCCCCCCCCC(=O)OC(C)C XUGNVMKQXJXZCD-UHFFFAOYSA-N 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000001788 mono and diglycerides of fatty acids Substances 0.000 description 1
- WQEPLUUGTLDZJY-UHFFFAOYSA-N n-Pentadecanoic acid Natural products CCCCCCCCCCCCCCC(O)=O WQEPLUUGTLDZJY-UHFFFAOYSA-N 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- OQCDKBAXFALNLD-UHFFFAOYSA-N octadecanoic acid Natural products CCCCCCCC(C)CCCCCCCCC(O)=O OQCDKBAXFALNLD-UHFFFAOYSA-N 0.000 description 1
- WWZKQHOCKIZLMA-UHFFFAOYSA-N octanoic acid Chemical compound CCCCCCCC(O)=O WWZKQHOCKIZLMA-UHFFFAOYSA-N 0.000 description 1
- 235000021313 oleic acid Nutrition 0.000 description 1
- 238000000614 phase inversion technique Methods 0.000 description 1
- IYDGMDWEHDFVQI-UHFFFAOYSA-N phosphoric acid;trioxotungsten Chemical compound O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.OP(O)(O)=O IYDGMDWEHDFVQI-UHFFFAOYSA-N 0.000 description 1
- 239000000955 prescription drug Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012488 sample solution Substances 0.000 description 1
- 238000009991 scouring Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011426 transformation method Methods 0.000 description 1
- 229940047183 tribulus Drugs 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
- A01N57/10—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds
- A01N57/14—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds containing aromatic radicals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/08—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing solids as carriers or diluents
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/22—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing ingredients stabilising the active ingredients
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/24—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing ingredients to enhance the sticking of the active ingredients
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P7/00—Arthropodicides
- A01P7/04—Insecticides
Landscapes
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Toxicology (AREA)
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Insects & Arthropods (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a method for preparing a high-drug-carrying phoxim nanostructure lipid carrier based on micro-flow control, which adopts an oil phase and a water phase which are respectively composed of components such as phoxim, solid lipid, liquid lipid, anionic surfactant, nonionic surfactant, cosurfactant, water and the like, and prepares the phoxim nanostructure lipid carrier in a micro-flow control system. The invention is favorable for preparing the phoxim nano-structured lipid carrier with environmental protection, high drug loading capacity, strong leaf surface residue capability, stable preparation, stable storage and good control effect, and realizes the efficient control of field lepidoptera pests such as prodenia litura.
Description
Technical Field
The invention belongs to the field of plant protection, and particularly relates to a method for preparing a high-drug-loading phoxim nanostructure lipid carrier based on microfluidics.
Background
Currently, the primary control method for field pests remains chemical pesticide control (Journal of hazardous materials,2020, 385:121525). Organophosphorus pesticides phoxim are used as a common traditional pesticide, the target of which is acetylcholinesterase of pests, but the effectiveness decreases with the increase of the application amount (Journal of Applied Entomology,2021,145 (5): 440-448). Phoxim has broad-spectrum toxicity to insects such as lepidoptera, and a representative pest is prodenia litura (Ningxia agricultural science and technology, 1982 (04): 40). The 1-2 instar prodenia litura larvae feed on a small amount of mesophyll, the feed intake of the 3 instar or more larvae is obviously increased, even the whole plant is destroyed, and meanwhile, the resistance of the large instar prodenia litura larvae to pesticides is enhanced, so that the large instar prodenia litura larvae are difficult to kill in the field (Biological Control,2020, 150:104348).
In order to effectively prevent and control prodenia litura and solve the problem that phoxim is insoluble in water, the prior patent and commodity medicine formula emulsifiable concentrates often contain a large amount of organic solvents (CN 110999900A; CN1729781; CN 102845461A), and the organic solvents in the phoxim emulsifiable concentrates are difficult to degrade in the environment and increase the danger of pesticide storage and transportation. Thus, the new formulation of phoxim should reduce the content of organic solvents or use degradable components while ensuring the foliar deposition and insecticidal capabilities of the pesticide.
The lipid carrier with nano structure can effectively improve the encapsulation efficiency of the medicine, enhance the slow release and transdermal capability of the medicine (Chinese journal of Chinese traditional medicine, 2017,42 (13): 2473-2478), and is expected to realize the reduction and synergy of pesticides. Nanostructured lipid carriers have found wide application in the pharmaceutical and cosmetic fields, due to their transdermal capabilities, while their use in the pesticide field is still at a relatively early stage. Most pesticides such as phoxim are lipophilic, so that the lipid can be used as a nano pesticide carrier material, and the nano structure lipid carrier is used for encapsulating phoxim, so that the adsorption capacity of pesticide blades can be improved, and the insecticidal capacity can be enhanced. Meanwhile, according to the principle of environmental friendliness, the surfactant in the structural lipid carrier raw material can be selected from natural nonionic surfactant or degradable anionic surfactant. Besides the composition of the components, the preparation method has important influence on the quality of the nanostructure lipid carrier.
The preparation method of the nanostructure lipid carrier comprises high-energy emulsification method and low-energy emulsification method (Nanoscale, 2021,13 (7): 4051-4059). The structural lipid carrier prepared by the high-energy emulsification method has smaller particle size and good stability, but the method has complex equipment and high energy consumption (Colloids and Surfaces A: physicochemical AND ENGINEERING ASPECTS,2020, 601:124982). Phase inversion is commonly used in low energy emulsification processes, but the process of preparation typically involves a drop-wise mixing of the oil and water phases, and thus the process is less controllable and stable (International journal of pharmaceutics,2014,477 (1-2): 208-217). The microfluidic method is a highly controllable preparation method of the nanostructure lipid carrier, and can realize high product quality, high heat transfer efficiency, automatic preparation, good batch processing repeatability and less external pollution (Nanoscale, 2021,13,19352-19366; nanoscales, 2019,11 (19): 9410-9421). The method can control the preparation of the nanostructure lipid carrier by adjusting the chip shape, the composition and the flow rate of the liquid in the channel (Langmuir, 2018,34 (13): 3961-3970;Chemical Engineering Science,2021,235:116468). The nano-structured lipid carrier prepared by the micro-flow control is mainly used for loading medical drugs, is not used for preparing pesticides, can be used for preparing high-drug-loading phoxim nano-structured lipid carrier by the micro-flow control, reduces the use of organic solvents in a formula, and is very important for efficient green control of lepidoptera insect pests such as prodenia litura.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a method for preparing a high-drug-carrying phoxim nanostructure lipid carrier based on microfluidics. The formula and the process in the preparation process are optimized, so that the environment friendliness of the phoxim pesticide is improved, the water accounts for more than 50%, and the raw materials are safe and degradable; the drug loading rate of the nano phoxim prepared in multiple batches reaches more than 98%, and the preparation stability is obviously improved; the prepared phoxim nanostructure lipid carrier has the advantages that the leaf surface retention rate is more than 62%, the semi-lethal concentration is as low as 3.16mg/L for 72 hours, the cold storage stability is more than 95%, the heat storage stability is more than 59%, the preparation method is superior to the original medicine and the commodity medicine, the prevention and control effect of phoxim is enhanced, and the high-quality and high-efficiency preparation of phoxim nanometer pesticide is realized.
In order to solve the problems in the prior art, the invention adopts the following technical scheme:
a method for preparing a high-drug-carrying phoxim nanostructure lipid carrier based on micro-flow control comprises the following steps:
Step 1, respectively preparing a water phase and an oil phase, wherein the water phase is formed by mixing a nonionic surfactant with water, and the oil phase is formed by mixing solid lipid, liquid lipid, phoxim, an anionic surfactant and a cosurfactant;
Step 2, in the microfluidic system, injecting the water phase and the oil phase into a side channel or a main channel of a connecting chip from the micro injection pump 1 and the micro injection pump 2 respectively, controlling the flow rate, mixing microfluid at the channel connection part of a heating platform, introducing the product into an ice water bath for cooling, and filtering through a filter membrane to form the phoxim nanostructure lipid carrier;
wherein the mass fraction of the water phase is 60-80%, and the mass fraction of the oil phase is 20-40%.
As an improvement, the mass fraction of the nonionic surfactant in the water phase is 70-90%, and the mass fraction of the nonionic surfactant is 10-30%.
As an improvement, the mass fraction of solid lipid in the oil phase is 1-10%, the mass fraction of liquid lipid is 10-30%, the mass fraction of phoxim is 30-60%, the mass fraction of anionic surfactant is 10-30%, and the mass fraction of cosurfactant is 10-30%.
As an improvement, the purity of the phoxim is 60-99%, the solid lipid is at least one of stearic acid, beeswax, monoglyceride, lecithin or cholesterol, the liquid lipid is at least one of oleic acid, palmitic acid, castor oil, isopropyl myristate, caprylic/capric glyceride or isopropyl palmitate, the anionic surfactant is at least one of lauryl polyoxyethylene ether sulfosuccinic acid monoester disodium salt (MES), lauryl sulfosuccinic acid monoester disodium salt (DLS), cocoyl monoethanolamide sulfosuccinic acid monoester disodium salt (DMSS) or lauramidopropyl hydroxysulfobetaine (LHSB-35), the nonionic surfactant is at least one of soybean saponin, tea saponin, tribulus saponin, ginsenoside or sea cucumber saponin, and the cosurfactant is at least one of ethanol, n-propanol, isopropanol, n-butanol or isobutanol.
As an improvement, the connecting chip of the microfluidic system is a T-shaped, Y-shaped or cross-shaped O/W-shaped or W/O-shaped chip.
As an improvement, the flow rates of the micro injection pump 1 and the micro injection pump 2 in the micro flow control system are respectively 20-40 mu L/min and 5-15 mu L/min, the temperature at the joint of the heating platform channel is 60-120 ℃, the cooling time in the ice water bath is 5-60min, and the aperture of the filter membrane is 0.1-1 mu m.
As an improvement, the drug loading rate of the phoxim nanostructure lipid carrier reaches more than 98 percent.
The beneficial effects are that:
Compared with the prior art, the method for preparing the high-drug-carrying phoxim nanostructure lipid carrier based on the microfluidics has the advantages that the water accounts for more than 50% by optimizing the formula and the process in the preparation process, and the raw materials are safe and degradable; the drug loading rate of the nano phoxim prepared in multiple batches reaches more than 98%, and the preparation stability is obviously improved; the prepared phoxim nanostructure lipid carrier has the advantages that the leaf surface retention rate is more than 62%, the semi-lethal concentration is as low as 3.16mg/L for 72 hours, the cold storage stability is more than 95%, the heat storage stability is more than 59%, the preparation method is superior to that of raw medicines and commodity medicines, the environment friendliness, the medicine carrying rate and the preparation stability of phoxim pesticides are improved, the prevention and treatment effect of phoxim is enhanced, and the high-quality and high-efficiency preparation of phoxim nanometer pesticides is realized. By the establishment of the technology, a new product and process are provided for the pesticide field, and an important reference basis is provided for realizing the reduction and synergy of the pesticide in the field.
Drawings
FIG. 1 is a schematic diagram of a microfluidic chip;
FIG. 2 is a schematic representation of the nanostructured lipid carrier phoxim prepared in example 1 of the present invention;
FIG. 3 is a schematic representation of a dry structured lipid carrier transmission electron microscope characterization;
FIG. 4 is an X-ray diffraction characterization of nanostructured lipid carriers and nano phoxim;
FIG. 5 is an infrared spectrum characterization of the drug substance, nanostructured lipid carrier and nano phoxim;
FIG. 6 is a Raman spectral characterization of nanostructured lipid carriers and nano phoxim;
Fig. 7 is a physical diagram of the microfluidic system used in example 1 of the present invention.
Detailed Description
The application will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present application and are not intended to limit the scope of the present application. Furthermore, it should be understood that various changes and modifications can be made by one skilled in the art after reading the teachings of the present application, and such equivalents are intended to fall within the scope of the application as defined in the appended claims.
Example 1
This example illustrates the microfluidic preparation of a high drug loaded phoxim nanostructure lipid carrier.
Example 1-1
The oil phase formula is solid lipid (monoglyceride), liquid lipid (oleic acid) and anionic surfactant (MES), and the water phase formula is water and nonionic surfactant (tea saponin). Different lipids (monoglyceride and oleic acid) -surfactants (MES and theasaponin) are arranged in a dual-phase system in a mass ratio of 1:9, 1:4, 3:7, 2:3 and 1:1, wherein the mass ratio of monoglyceride to oleic acid in the lipids is 1:1, the mass ratio of anionic surfactant MES and nonionic surfactant theasaponin in the surfactants is 1:1, and the mass fraction of water is 70%. Then, 10% of cosurfactant n-propanol and 2% of phoxim with total biphasic mass fraction are additionally added into the oil phase as shown in table 1.
Wherein, the purity of monoglyceride is 90%, the purity of phoxim is 99%, the purity of MES is 30%, the purity of tea saponin is 95%, and the rest components are all analytically pure.
In a microfluidic system, the flow rates of the microinjection pumps 1 and 2 were controlled to be 22 to 27. Mu.L/min and 6 to 13. Mu.L/min, respectively (Environmental SCIENCE AND Pollution Research, 2022:1-13.), as shown in Table 1. The water phase and the oil phase are respectively injected into a side channel and a main channel of the T-shaped connecting chip, and microfluid is mixed at the joint of the channels with the temperature of 85 ℃ of the heating platform. And then the product is introduced into ice water bath to be cooled for 30min, and filtered through a 0.22 mu m filter membrane to form the lipid carrier phoxim with the nano structure.
100. Mu.L of the nano-pesticide was mixed with 600. Mu.L of n-hexane and turned upside down 10 times for extraction of the non-entrapped phoxim into n-hexane. Subsequently, 100. Mu.L of the extract was aspirated into a 2mL centrifuge tube, the extract was dried and 400. Mu.L of methanol was supplemented, and the concentration of phoxim at C 1 was determined. On the other hand, 100. Mu.L of the nano-pesticide was mixed with 500. Mu.L of methanol, left to stand for 30 minutes until the loaded phoxim was dissolved, and the total phoxim concentration in the mixture was determined to be C 2.
The Encapsulation Efficiency (EE) of the nanostructured lipid carrier for phoxim was calculated as follows: ee= (C 2-C1)/C2 x 100%) in the formula, EE is encapsulation efficiency, C 1 is concentration of non-entrapped phoxim, and C 2 is concentration of total phoxim, resulting in drug loading rate.
TABLE 1 Effect of lipid-surfactant ratios on encapsulation efficiency
As can be seen from table 1, as the ratio of the lipid surfactant increases, the encapsulation efficiency of phoxim decreases; EE drops significantly to 97.79±0.16% at a ratio of 1:1. In order to ensure the encapsulation efficiency of phoxim and reduce the dosage of the surfactant, the ratio of the lipid to the surfactant is determined to be better when the ratio is 1:4.
Examples 1 to 2
The oil phase formula is solid lipid (monoglyceride), liquid lipid (oleic acid) and anionic surfactant (MES), the water phase formula is water and nonionic surfactant (tea saponin), the mass ratio of 1:4 of lipid (monoglyceride and oleic acid) to surfactant (MES and tea saponin) in the biphasic system is selected, the mass ratio of MES and tea saponin in the surfactant is 1:1, the mass ratio of different solid lipid (monoglyceride) to liquid lipid (oleic acid) is set to be 2:1, 1:1, 1:2, 1:3, 1:4 and 1:5, and the mass fraction of water is 70%. Then, 10% of cosurfactant n-propanol and 2% of phoxim with total biphasic mass fraction were additionally added into the oil phase as shown in table 2.
The component purity, microfluidic system preparation parameters, and encapsulation efficiency test methods were as described above, and are shown in table 2.
TABLE 2 Effect of solid-liquid lipid ratio on encapsulation efficiency
As can be seen from Table 2, the encapsulation efficiency increased with decreasing solid-liquid lipid ratio, and was 99.35.+ -. 0.04% at a maximum value of 1:4. However, when the ratio is reduced to 1:5, the encapsulation efficiency is significantly reduced. Therefore, the solid-liquid lipid ratio is preferably 1:4.
Examples 1 to 3
The oil phase formula is set to be solid lipid (monoglyceride), liquid lipid (oleic acid) and anionic surfactant (MES), the water phase formula is set to be water and nonionic surfactant (tea saponin), the mass ratio of 1:4 of lipid (monoglyceride and oleic acid) to surfactant (MES and tea saponin) in the two-phase system is selected to be 1:4, the mass ratio of solid lipid (monoglyceride) to liquid lipid (oleic acid) is set to be 1:3, 1:1, 3:1, 5:1 and 7:1, and the mass ratio of different anionic surfactants (MES) to nonionic surfactant (tea saponin) is set to be 1:3, 1:1, 5:1 and 7:1, and the mass ratio of water is 70%. Then, 10% of cosurfactant n-propanol and 2% of phoxim with total biphasic mass fraction were additionally added into the oil phase as shown in table 3.
The component purity, microfluidic system preparation parameters, and encapsulation efficiency test methods were as described above, and are shown in table 3.
TABLE 3 influence of anionic-nonionic surfactant ratio on encapsulation efficiency
As can be seen from Table 3, the increase in the anionic-nonionic surfactant ratio increases the encapsulation efficiency of phoxim, which is nearly 100%, and the preferred ratio is determined to be 1:3 due to the cost of the theasaponin.
Examples 1 to 4
The oil phase formula is set to be solid lipid (monoglyceride), liquid lipid (oleic acid) and anionic surfactant (MES), the water phase formula is water and nonionic surfactant (tea saponin), the mass ratio of 1:4 of lipid (monoglyceride and oleic acid) to surfactant (MES and tea saponin) in the two-phase system is selected to be 1:4, the mass ratio of solid lipid (monoglyceride) to liquid lipid (oleic acid) is selected to be 1:3, and the mass ratio of anionic surfactant (MES) to nonionic surfactant (tea saponin) is selected to be 70 percent. Then, the cosurfactant n-propanol with the total mass fraction of 10% is additionally added into the oil phase, and the phoxim with different mass fractions of 2%, 4%, 6%, 8% and 10% are respectively added, as shown in table 4.
The component purity, microfluidic system preparation parameters, and encapsulation efficiency test methods were as described above, and are shown in table 4.
TABLE 4 influence of phoxim addition on encapsulation efficiency
As can be seen from table 4, when the addition amount of phoxim was increased from 2% to 4%, the encapsulation efficiency of phoxim was significantly reduced from 98.94±0.11% to 97.52 ±0.58%; however, after addition of 10% phoxim, the encapsulation efficiency increased significantly to 99.02±0.10%. Meanwhile, in the present system, the lipid content in the nanostructured lipid carrier is 6%, and the addition amount of phoxim exceeding 10% exceeds the loading capacity of the system, so that the addition amount of phoxim is determined to be preferably 10%.
Examples 1 to 5
The oil phase formula is set to be solid lipid (monoglyceride), liquid lipid (oleic acid) and anionic surfactant (MES), the water phase formula is water and nonionic surfactant (tea saponin), the mass ratio of 1:4 of lipid (monoglyceride and oleic acid) to surfactant (MES and tea saponin) in the two-phase system is selected to be 1:4, the mass ratio of solid lipid (monoglyceride) to liquid lipid (oleic acid) is selected to be 1:3, and the mass ratio of anionic surfactant (MES) to nonionic surfactant (tea saponin) is selected to be 70 percent. After that, 10% of phoxim by mass of the total diphase is additionally added into the oil phase, and 0%,5%,10%,15% and 20% of cosurfactant n-propanol with different mass fractions are added respectively, as shown in table 5.
The component purity, microfluidic system preparation parameters, and encapsulation efficiency test methods were as described above, and are shown in table 5.
TABLE 5 Effect of cosurfactant n-propanol addition on encapsulation efficiency
As can be seen from table 5, the encapsulation efficiency of the nanostructured lipid carrier decreased with increasing co-surfactant n-propanol addition, the maximum encapsulation efficiency of the nanostructured lipid carrier without co-surfactant was 99.02±0.03%, and the encapsulation efficiency was significantly reduced to 87.26 ±0.30% and 73.38±2.74% when the co-surfactant addition was 15% and 20%, respectively. However, when no cosurfactant was added, it was found in the experiment that there was an insoluble matter in the oil phase, and therefore, it was preferable to select to add 5% of cosurfactant.
Example 2
This example illustrates the stability of microfluidic preparation of high drug loaded phoxim nanostructure lipid carriers.
Microfluidic preparation:
biphase formulation (components in parts by mass): 74.8% of water phase and 25.2% of oil phase;
Aqueous phase formula (components in parts by mass): 79.55% of water and 20.45% of tea saponin;
oil phase formula (components in parts by mass): 39.68% of phoxim, 4.05% of monoglyceride, 16.19% of oleic acid, 20.24% of MES and 19.84% of n-propanol.
The component purity and microfluidic system preparation parameters were the same as in example 1.
And (3) preparing by a phase transformation method: the oil and water phases of the above formulation were placed in test tubes and heated at 85 ℃ in an oil bath magnetic stirrer at a speed of 200 rpm. Then, the whole of the aqueous phase was added dropwise to the oily phase and stirred for 30 minutes. Finally, the mixture was ice-water-bath for 30 minutes and passed through a 0.22 μm filter as a phase inversion sample.
The encapsulation efficiency of the nano pesticide prepared by the two methods is respectively measured, and the encapsulation efficiency test method is the same as that of example 1.
And calculating the standard error value of the encapsulation efficiency of the nano pesticide prepared for 10 times, and judging the batch stability of the two preparation methods.
Table 6 encapsulation efficiency of 10 batches of nano pesticides prepared by microfluidic and phase change methods
As shown in table 6, the encapsulation rates of 10 batches of nano pesticides prepared by the microfluidic and phase change methods were calculated with average values of 98.67 ±0.47% and 46.01±12.46%, respectively; the standard error of the encapsulation efficiency of the nano pesticide prepared by the microfluidic method is far smaller than that of the nano pesticide prepared by the phase inversion method, which indicates that the nano pesticide prepared by the method has higher batch stability.
Example 3
This example illustrates the comparison of green high drug loading nano phoxim pesticide to phoxim leaf surface affinity of other dosage forms.
The lipid carrier with the phoxim nanostructure was prepared by the microfluidic method as described in example 2.
The method for preparing the phoxim spraying raw material comprises the following steps of: 100 mug of phoxim with the purity of 99 percent is mixed with 2mL of acetone to form a standard solution, and the standard solution is diluted with water according to the required concentration; the phoxim content of the phoxim commercial formulation was 40% and was purchased from the eastern pesticide chemical plant (thailand, china).
The dilution concentration of the three formulations is 100mg/L phoxim concentration, the diameter of the cultivated mulberry seedling is 80+/-20 mm, the height is 20-30 cm, and the plants are used for detention and anti-scouring tests. The fresh mulberry leaves were then perforated (3 cm diameter) with a punch and weighed to m 1. Then, the leaf disc formed by punching was immersed in the aqueous pesticide solution having the same concentration as above for 15s, the leaf disc was taken out with forceps, left for 30s, and weighed to m 2. Three dosage forms were each assayed and repeated 10 times. The calculation formula of the leaf surface retention of the pesticide is as follows: r= (m 2-m1)/S, where R is the retention, m 1 is the mass of untreated leaf disks, m 2 is the mass of treated leaf disks, and S is the leaf disk area.
Spraying 1ml of pesticide diluent on the mulberry leaves, and naturally airing for 2 hours. Pure water was added dropwise to the mulberry leaves at a rate of 10 ml/min for 8 minutes to simulate rain wash. The leaves were punched out, minced and collected into centrifuge tubes containing 5 ml methanol. The tube was sonicated at 200W for 10min with 5s for both the on time and the off time. Thereafter, the tube was centrifuged at 12000rpm for 3min, 1mL of the supernatant was aspirated and mixed with an equal amount of methanol as a sample. Finally, the concentration of phoxim in the sample was measured, the residual rate was calculated, and the residual rate was calculated from the percentage of the concentration at the time point and the concentration at the time of 0min of rinsing.
TABLE 7 foliar affinity of phoxim sprayed drug substance, phoxim commercial formulation and phoxim nanostructured lipid carrier
As shown in table 7, the leaf retention of the commercial formulation of phoxim was significantly lower than that of the phoxim sprayed drug substance and phoxim nanostructured lipid carrier. The foliar retention makes the phoxim nanostructure lipid carrier easier to be ingested by pests, and reduces the waste caused by rain wash.
Example 4
This example illustrates the comparison of toxicity of green high drug loaded nano phoxim pesticide with other formulations of phoxim to prodenia litura.
The phoxim spray stock, phoxim commercial formulation, phoxim nanostructure lipid carrier and leaf discs were obtained as described in example 3.
Bioassays assessed toxicity of different phoxim formulations and theasaponin to prodenia litura using a leaf dipping method. The dilution concentration of the treatment is 1-50g/L of theasaponin, 10-20mg/L of phoxim sprayed original drug, 10-50mg/L of phoxim commercial prescription drug and 2.5-12.5mg/L of phoxim nanostructure lipid carrier. Sang Sheshe dishes (3 cm diameter) were immersed in the diluent for 30s and then dried.
The larvae were fed manually in a reproduction room at 25.+ -. 1 ℃ and relative humidity of 60-70% with light/dark cycle of 16/8 h. Larvae at 3 days of 3 years were selected for pesticide toxicity determination. Thereafter, leaf dishes and 24h starved larvae were placed in 35mm agar-bearing plastic dishes, with the other conditions being the same. Larval mortality was calculated after 24h, 48h and 72h treatment, and the concentration of phoxim was calculated for the semi-lethal concentration LC 50 of prodenia litura in different formulations by means of the probit regression model.
Table 8 toxicity of phoxim sprayed drug substance, phoxim commercial formulation and phoxim nanostructured lipid carrier
Three formulations as shown in table 8 were run on the probit model of prodenia litura within 72 hours. At 24h, the standard LC 50 of the crude drug sprayed with phoxim, the commercial formulation of phoxim and the lipid carrier with phoxim nano structure is 20.76, 78.29 and 6.39mg/L respectively, and at 72h LC 50 is continuously reduced to 19.90, 70.67 and 3.16mg/L. Under the semi-lethal concentration, the dosage of the phoxim nanostructure lipid carrier is 15.9 to 30.8 percent of the original drug sprayed by phoxim, and 4.5 to 8.2 percent of the commercial formulation drug of phoxim. Meanwhile, in the embodiment, the prodenia litura larvae treated by 10-50 g/L of tea saponin and 0-10 mg/L of nano-structured lipid carrier (the same preparation method as that of the phoxim nano-structured lipid carrier is except that phoxim is removed) do not die after 24 hours, which indicates that the toxicity of the phoxim nano-structured lipid carrier is not influenced by the carrier.
Example 5
This example illustrates the comparison of cold and hot storage properties of green high drug loading nano phoxim pesticides and phoxim of other dosage forms.
Firstly, diluting the original pesticide sprayed by phoxim, the commercial phoxim and the phoxim nano-drug to 100mg/L phoxim concentration, and pouring 10mL of diluent into a test tube. In the cold storage experiment, 200 mu L of sample solution is firstly sampled, the initial phoxim concentration is detected by using liquid chromatography, and then the residual diluent is placed in a refrigerator at 0 ℃ for cold storage for 15 days, and the concentration of the phoxim in the sample is measured, namely the residual phoxim concentration. The heat storage experiment is basically the same as that of the sample heat storage method, namely, the sample is placed in a water bath kettle with the temperature of 54 ℃ for heat storage for 15d. And calculating the residual rate of the phoxim of the sample after cold storage and hot storage treatment, wherein the residual rate is the percentage of the residual concentration of the phoxim to the original concentration.
Table 9 cold and heat storage capacities of phoxim sprayed drug substance, phoxim commercial formulation and phoxim nanostructured lipid carrier
As shown in Table 9, the residual rates of the three dosage forms of phoxim after 14d of cold storage of the pesticide are 52.21 +/-18.21, 25.84+/-4.60 and 95.23+/-3.90 percent respectively, and the cold storage residual rate of the phoxim nano-drug is obviously higher than that of the other two dosage forms.
The residual rates of the three dosage forms after 14d of heat storage of the pesticide are 19.12+/-1.36, 27.61+/-8.95 and 59.30+/-16.35 percent respectively, and the heat storage residual rate of the phoxim nano-drug is obviously higher than that of the other two dosage forms.
The cold storage stability and the hot storage stability of the pesticide are evaluation indexes for analyzing whether the pesticide can be decomposed in storage, the cold storage stability of the phoxim nano-drug is higher, the hot storage stability of the phoxim nano-drug is effectively improved by replacing a carrier or concentrating components, and the cold storage stability and the hot storage stability provide a foundation for long-term storage and practical application of the phoxim nano-drug.
Example 6
This example illustrates the characterization of a green high drug loading nano phoxim pesticide in relation to other dosage forms of phoxim.
The structural schematic diagram of the nano-structured lipid carrier phoxim prepared by the invention is shown in figure 2.
(1) Particle size and potential characterization
The particle size and Zeta potential of the phoxim nanostructure lipid carrier, the phoxim sprayed active compound and the phoxim commercial formulation are measured by a multi-angle particle size analyzer. All three formulations were diluted 10-fold to 10mL with water and the instrument operating temperature was 25 ℃.
Table 10 particle size and Zeta potential of phoxim sprayed drug substance, phoxim commercial formulation and phoxim nanostructured lipid carrier
The particle sizes and Zeta potentials of the three formulations are shown in Table 10. The particle sizes of the three dosage forms are 768.58 +/-27.47, 362.78+/-1.29 and 120.08+/-0.65 nm respectively, and the particle size of the phoxim nanostructure lipid carrier is greatly reduced. The Zeta potentials of the three dosage forms are-0.77+/-0.32, 0.61+/-0.24 and-39.61 +/-1.33 mV respectively, and the absolute value of the potential of the phoxim nanostructure lipid carrier is larger and more stable.
(2) Particle morphology characterization
Particle morphology as shown in figure 3, particle morphology of the phoxim nanostructure lipid carrier is characterized by a Transmission Electron Microscope (TEM), a preparation method of a TEM sample of the phoxim nanostructure lipid carrier is that the sample is diluted by adding water for 10 times, 1 drop of diluted liquid is sucked by a 10 mu L liquid-transferer and added on a copper mesh covered with a carbon film for standing for 5min, the copper mesh is naturally dried after the excessive liquid is sucked by filter paper, 2% phosphotungstic acid is continuously dripped on the dried copper mesh for 1min, and then the excessive liquid is sucked by the filter paper and the copper mesh is dried to be used as a TEM detection sample.
(3) Crystal form and chemical bond characterization
The samples used in this experiment were all powders, and the treatment method was: and respectively adding 5 times of n-hexane into the samples of the blank nanostructure lipid carrier and the phoxim nanostructure lipid carrier, reversely mixing, discarding the n-hexane, repeatedly cleaning for 3 times, volatilizing the n-hexane remained in the samples, and freeze-drying for 48 hours to obtain powder blank nanostructure lipid carrier and phoxim nanostructure lipid carrier samples, wherein the powder samples are also used for infrared and Raman characterization experiments. The components and crystal forms of the blank nanostructure lipid carrier and the phoxim nanostructure lipid carrier are detected by an X-ray diffraction instrument, the scanning speed is 2 degrees/min, and the scanning angle is 5-90 degrees.
The X-ray diffraction characterization is shown in figure 4, and the X-ray diffraction XRD results of the blank nano-structured lipid carrier and the phoxim nano-structured lipid carrier comprise monoclinic phases of glyceryl monostearate GMS (JCPDS file number 08-0590) and oleic acid (JCPDS file number 37-1811), which shows that the blank NLC contains GMS and oleic acid, and the P-NLC loaded with phoxim also contains the two lipids, and the P-NLC is proved to be the nano-structured lipid carrier. The broad impurity peaks indicate that both the crystalline forms of the blank NLC and P-NLC are quasi-amorphous.
Whether phoxim is successfully encapsulated into a blank nanostructured lipid carrier and the chemical bond change of the phoxim nanostructured lipid carrier component is determined by fourier transform infrared spectroscopy, FTIR. The phoxim original drug used in the infrared spectrum experiment does not contain acetone, and the treatment method of the blank nano-structured lipid carrier and the phoxim nano-structured lipid carrier is the same as the above.
The fourier infrared spectrum is characterized in that as shown in fig. 5, the phoxim-related absorption peak also appears in the phoxim nanostructure lipid carrier, proving that phoxim has been successfully loaded into the nanostructure lipid carrier.
Raman spectroscopy was used to verify changes in the lipid carrier chemical bonds of phoxim nanostructures. The treatment method of the blank nanostructured lipid carrier and the phoxim nanostructured lipid carrier is the same as the above.
The Raman spectrum characterization is shown in figure 6, the nanostructure lipid carrier has no obvious characteristic peak, and the phoxim nanostructure lipid carrier contains characteristic peak. The relevant characteristic peak of the P-NLC in the Raman spectrum indicates that phoxim has been successfully loaded into the NLC.
In summary, by optimizing the formula and the process in the preparation process, the water accounts for more than 50%, and the raw materials are safe and degradable; the drug loading rate of the nano phoxim prepared in multiple batches reaches more than 98%, and the preparation stability is obviously improved; the prepared phoxim nanostructure lipid carrier has the advantages that the leaf surface retention rate is more than 62%, the semi-lethal concentration is as low as 3.16mg/L for 72 hours, the cold storage stability is more than 95%, the heat storage stability is more than 59%, the preparation method is superior to that of raw medicines and commodity medicines, the environment friendliness, the medicine carrying rate and the preparation stability of phoxim pesticides are improved, the prevention and treatment effect of phoxim is enhanced, and the high-quality and high-efficiency preparation of phoxim nanometer pesticides is realized. By the establishment of the technology, a new product and process are provided for the pesticide field, and an important reference basis is provided for realizing the reduction and synergy of the pesticide in the field.
The foregoing is only a preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art, who is within the scope of the present invention, should be covered by the protection scope of the present invention by making equivalents and modifications to the technical solution and the inventive concept thereof.
Claims (2)
1. The method for preparing the high-drug-carrying phoxim nanostructure lipid carrier based on the microfluidics is characterized by comprising the following steps of:
Step 1, respectively preparing a water phase and an oil phase, wherein the water phase is formed by mixing nonionic surfactant tea saponin with water, and the oil phase is formed by mixing solid lipid monoglyceride, liquid lipid oleic acid, phoxim, anionic surfactant lauryl polyoxyethylene ether sulfosuccinic monoester disodium salt and cosurfactant n-propanol; wherein the components in parts by weight are as follows: 74.8% of water phase and 25.2% of oil phase; the total system mass part of water in the water phase is 79.55%, and the nonionic surfactant theasaponin is 20.45%; the total system mass fraction of solid lipid monoglyceride in the oil phase is 4.05%, the mass fraction of liquid lipid oleic acid is 16.19%, the mass fraction of phoxim is 39.68%, the mass fraction of anionic surfactant laureth sulfosuccinic acid monoester disodium salt is 20.24%, and the mass fraction of cosurfactant n-propanol is 19.84%;
And 2, injecting the water phase and the oil phase into a side channel or a main channel of a T-shaped connecting chip from a micro injection pump 1 and a micro injection pump 2 respectively in a micro flow control system, controlling the flow rate, mixing micro fluid at the channel connection part of a heating platform, introducing the product into an ice water bath for cooling, and filtering through a filter membrane to form the phoxim nano structure lipid carrier, wherein the flow rates of the micro injection pump 1 and the micro injection pump 2 in the micro flow control system are 26.67 mu L/min and 8.18 mu L/min respectively, the temperature at the channel connection part of the heating platform is 85 ℃, the cooling time in the ice water bath is 30 min, and the pore diameter of the filter membrane is 0.22 mu m.
2. The method for preparing the high-drug-carrying phoxim nanostructure lipid carrier based on the micro-flow control of claim 1, wherein the drug-carrying rate of the phoxim nanostructure lipid carrier is more than 98%.
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| 微流控技术在纳米药物载体制备中的应用;孙漩嵘 等;《中国药学杂志》;第55卷(第8期);第573-579页 * |
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