CN112675077A - Water-soluble microemulsion and preparation method thereof - Google Patents
Water-soluble microemulsion and preparation method thereof Download PDFInfo
- Publication number
- CN112675077A CN112675077A CN202110013004.3A CN202110013004A CN112675077A CN 112675077 A CN112675077 A CN 112675077A CN 202110013004 A CN202110013004 A CN 202110013004A CN 112675077 A CN112675077 A CN 112675077A
- Authority
- CN
- China
- Prior art keywords
- water
- microemulsion
- soluble
- surfactant
- soluble microemulsion
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000004530 micro-emulsion Substances 0.000 title claims abstract description 99
- 238000002360 preparation method Methods 0.000 title claims description 21
- 238000000593 microemulsion method Methods 0.000 title description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 34
- 239000004094 surface-active agent Substances 0.000 claims abstract description 22
- 239000011487 hemp Substances 0.000 claims abstract description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 244000025254 Cannabis sativa Species 0.000 claims abstract description 14
- 235000012766 Cannabis sativa ssp. sativa var. sativa Nutrition 0.000 claims abstract description 14
- 235000012765 Cannabis sativa ssp. sativa var. spontanea Nutrition 0.000 claims abstract description 14
- 235000009120 camo Nutrition 0.000 claims abstract description 14
- 235000005607 chanvre indien Nutrition 0.000 claims abstract description 14
- 238000001228 spectrum Methods 0.000 claims abstract description 14
- 241000186427 Cutibacterium acnes Species 0.000 claims abstract description 11
- ZTGXAWYVTLUPDT-UHFFFAOYSA-N cannabidiol Natural products OC1=CC(CCCCC)=CC(O)=C1C1C(C(C)=C)CC=C(C)C1 ZTGXAWYVTLUPDT-UHFFFAOYSA-N 0.000 claims abstract description 11
- 229940055019 propionibacterium acne Drugs 0.000 claims abstract description 11
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 10
- QHMBSVQNZZTUGM-UHFFFAOYSA-N Trans-Cannabidiol Natural products OC1=CC(CCCCC)=CC(O)=C1C1C(C(C)=C)CCC(C)=C1 QHMBSVQNZZTUGM-UHFFFAOYSA-N 0.000 claims abstract description 10
- QHMBSVQNZZTUGM-ZWKOTPCHSA-N cannabidiol Chemical compound OC1=CC(CCCCC)=CC(O)=C1[C@H]1[C@H](C(C)=C)CCC(C)=C1 QHMBSVQNZZTUGM-ZWKOTPCHSA-N 0.000 claims abstract description 10
- 229950011318 cannabidiol Drugs 0.000 claims abstract description 10
- PCXRACLQFPRCBB-ZWKOTPCHSA-N dihydrocannabidiol Natural products OC1=CC(CCCCC)=CC(O)=C1[C@H]1[C@H](C(C)C)CCC(C)=C1 PCXRACLQFPRCBB-ZWKOTPCHSA-N 0.000 claims abstract description 10
- 229910001868 water Inorganic materials 0.000 claims abstract description 10
- 239000004064 cosurfactant Substances 0.000 claims abstract description 8
- 239000000203 mixture Substances 0.000 claims description 18
- 239000003921 oil Substances 0.000 claims description 17
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 230000003266 anti-allergic effect Effects 0.000 claims description 12
- 239000002537 cosmetic Substances 0.000 claims description 12
- 230000002000 scavenging effect Effects 0.000 claims description 12
- 229920001223 polyethylene glycol Polymers 0.000 claims description 11
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- -1 superoxide anions Chemical class 0.000 claims description 9
- 238000009472 formulation Methods 0.000 claims description 8
- 230000002401 inhibitory effect Effects 0.000 claims description 8
- 230000003110 anti-inflammatory effect Effects 0.000 claims description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 5
- 239000012498 ultrapure water Substances 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 4
- 241000218236 Cannabis Species 0.000 claims description 3
- 239000002202 Polyethylene glycol Substances 0.000 claims description 3
- 239000004359 castor oil Substances 0.000 claims description 3
- 235000019438 castor oil Nutrition 0.000 claims description 3
- 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 claims description 3
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 3
- 229920000053 polysorbate 80 Polymers 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- 210000003491 skin Anatomy 0.000 abstract description 19
- 230000000694 effects Effects 0.000 abstract description 16
- 108010003272 Hyaluronate lyase Proteins 0.000 abstract description 12
- 102000001974 Hyaluronidases Human genes 0.000 abstract description 12
- 229960002773 hyaluronidase Drugs 0.000 abstract description 12
- 206010018910 Haemolysis Diseases 0.000 abstract description 8
- 239000002245 particle Substances 0.000 abstract description 8
- 230000028327 secretion Effects 0.000 abstract description 8
- 108090001007 Interleukin-8 Proteins 0.000 abstract description 7
- 102000004890 Interleukin-8 Human genes 0.000 abstract description 7
- 206010061218 Inflammation Diseases 0.000 abstract description 6
- 210000002510 keratinocyte Anatomy 0.000 abstract description 6
- 230000004054 inflammatory process Effects 0.000 abstract description 3
- 230000015271 coagulation Effects 0.000 abstract description 2
- 238000005345 coagulation Methods 0.000 abstract description 2
- 230000012010 growth Effects 0.000 abstract description 2
- 230000032798 delamination Effects 0.000 abstract 1
- 230000007774 longterm Effects 0.000 abstract 1
- 238000009827 uniform distribution Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 22
- 230000005764 inhibitory process Effects 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 238000002835 absorbance Methods 0.000 description 14
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 11
- 210000003743 erythrocyte Anatomy 0.000 description 8
- 235000011187 glycerol Nutrition 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 6
- 230000008588 hemolysis Effects 0.000 description 6
- WQGWDDDVZFFDIG-UHFFFAOYSA-N pyrogallol Chemical compound OC1=CC=CC(O)=C1O WQGWDDDVZFFDIG-UHFFFAOYSA-N 0.000 description 6
- 150000003254 radicals Chemical class 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 5
- 239000008367 deionised water Substances 0.000 description 5
- 229910021641 deionized water Inorganic materials 0.000 description 5
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 4
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 239000013067 intermediate product Substances 0.000 description 3
- 229940079877 pyrogallol Drugs 0.000 description 3
- 239000012488 sample solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 239000012980 RPMI-1640 medium Substances 0.000 description 2
- CYQFCXCEBYINGO-UHFFFAOYSA-N THC Natural products C1=C(C)CCC2C(C)(C)OC3=CC(CCCCC)=CC(O)=C3C21 CYQFCXCEBYINGO-UHFFFAOYSA-N 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 230000004888 barrier function Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- CYQFCXCEBYINGO-IAGOWNOFSA-N delta1-THC Chemical compound C1=C(C)CC[C@H]2C(C)(C)OC3=CC(CCCCC)=CC(O)=C3[C@@H]21 CYQFCXCEBYINGO-IAGOWNOFSA-N 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 229960004242 dronabinol Drugs 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 235000019441 ethanol Nutrition 0.000 description 2
- 230000007760 free radical scavenging Effects 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- BGNGWHSBYQYVRX-UHFFFAOYSA-N 4-(dimethylamino)benzaldehyde Chemical compound CN(C)C1=CC=C(C=O)C=C1 BGNGWHSBYQYVRX-UHFFFAOYSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 206010012335 Dependence Diseases 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 241000255925 Diptera Species 0.000 description 1
- 101001055222 Homo sapiens Interleukin-8 Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- 206010023126 Jaundice Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 206010040880 Skin irritation Diseases 0.000 description 1
- 229920002385 Sodium hyaluronate Polymers 0.000 description 1
- 102100029613 Transient receptor potential cation channel subfamily V member 1 Human genes 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000002921 anti-spasmodic effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 238000006701 autoxidation reaction Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229930003827 cannabinoid Natural products 0.000 description 1
- 239000003557 cannabinoid Substances 0.000 description 1
- 229940065144 cannabinoids Drugs 0.000 description 1
- 229960003453 cannabinol Drugs 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000812 cholinergic antagonist Substances 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 238000004134 energy conservation Methods 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 150000003278 haem Chemical class 0.000 description 1
- 230000003400 hallucinatory effect Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000002949 hemolytic effect Effects 0.000 description 1
- 239000005457 ice water Substances 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 210000005007 innate immune system Anatomy 0.000 description 1
- 238000000464 low-speed centrifugation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000036556 skin irritation Effects 0.000 description 1
- 231100000475 skin irritation Toxicity 0.000 description 1
- 229940010747 sodium hyaluronate Drugs 0.000 description 1
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
- SWGJCIMEBVHMTA-UHFFFAOYSA-K trisodium;6-oxido-4-sulfo-5-[(4-sulfonatonaphthalen-1-yl)diazenyl]naphthalene-2-sulfonate Chemical compound [Na+].[Na+].[Na+].C1=CC=C2C(N=NC3=C4C(=CC(=CC4=CC=C3O)S([O-])(=O)=O)S([O-])(=O)=O)=CC=C(S([O-])(=O)=O)C2=C1 SWGJCIMEBVHMTA-UHFFFAOYSA-K 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Images
Landscapes
- Cosmetics (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a water-soluble microemulsion, which is characterized by comprising cannabidiol, a surfactant, a cosurfactant and water, wherein the microemulsion comprises the following components in percentage by weight: 0.5-5 wt% of industrial hemp full-spectrum oil; PPG-13-decyltetradecanol polyether-242-40 wt%; 2-40 wt% of PEG/PPG/polytetramethylene glycol-8/5/3 glycerol, and the balance of water. The microemulsion of the invention has clear and bright color, uniform distribution of droplets with the particle size of about 10-30 nm, good thermodynamic stability and no delamination after long-term storage. Meanwhile, the microemulsion can effectively inhibit the activity of hyaluronidase, inhibit the secretion of IL-8 of human keratinocytes and promote the generation of IL-10, remarkably inhibit the growth of staphylococcus aureus and Propionibacterium acnes, remove free radicals, and cannot generate obvious coagulation or hemolytic reaction, so that the inflammatory reaction of skin is relieved, and the anti-inflammation and the relief are realized.
Description
Technical Field
The invention belongs to the technical field of daily cosmetics, and particularly relates to a water-soluble microemulsion and a preparation method thereof.
Background
The skin serves as the first defense barrier of the innate immune system and one of the main functions is to protect the body from external agents. However, as society develops and the ecological environment deteriorates, the skin is challenged more and more. For example, mosquitoes abuse or a large amount of dust easily causes red swelling of the skin of people, and in severe cases, dermatitis, allergy and local hemonecrosis are caused, so that bacteria are enriched on the surface of the skin.
Cannabidiol (CBD) is one of the main components of cannabinoids, has no side effects of addiction and hallucinogenic of Tetra-Hydro Cannabinol (THC), has the functions of anti-inflammation, antibiosis, pain relief, antioxidation, antispasmodic, antitumor and the like, and is mainly related to central cannabis receptor CB1, peripheral cannabis receptor CB2 and capsaicin receptor TRPV1 in pharmacology. With the updating of regulations and the continuous and intensive research on the application functions thereof, CBD at low concentration is beginning to emerge in health care products, food products and skin care products.
The effective components of the cosmetic are partially absorbed by the skin, which determines the quality of the cosmetic. However, the active molecules in cosmetics are complex in structure and low in solubility, and thus effective concentration on skin is difficult to achieve. CN201310068002.X discloses a cannabinol compound microemulsion and a preparation method thereof, but in the prior art, the existence of an oil phase enables the microemulsion to have a large particle size, which is not beneficial to skin permeation and absorption.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the water-soluble microemulsion which effectively permeates skin with small particle size, relieves skin irritation and lightens skin inflammatory reaction.
The invention provides a water-soluble microemulsion which is characterized by comprising cannabidiol, a surfactant, a cosurfactant and water.
Further, in the water-soluble microemulsion, the cannabidiol is selected from: the industrial hemp full-spectrum oil contains cannabidiol and is 45-55 wt%.
Further, in the water-soluble microemulsion, the surfactant is selected from PPG-13-decyltetradecanol polyether-24, Tween 80, PEG40 hydrogenated castor oil.
Further, in the water-soluble microemulsion, the co-surfactant is selected from PEG/PPG/polytetramethylene glycol-8/5/3 glycerol, propylene glycol, isopropanol, polyethylene glycol, glycerol.
Further, in the water-soluble microemulsion, the weight percentages of the industrial hemp full-spectrum oil are: 0.5 to 5 wt%, preferably 0.5 to 2 wt%, and more preferably 0.5 to 1 wt% of the total weight of the water-soluble microemulsion.
Further, in the water-soluble microemulsion, the weight percentage of the surfactant is: 2-40 wt%, preferably 2-30 wt%, more preferably 2-7 wt% of the total weight of the water-soluble microemulsion.
Further, in the water-soluble microemulsion, the weight percentage of the cosurfactant is as follows: 2-40 wt%, preferably 2-30 wt%, more preferably 2-7 wt% of the total weight of the water-soluble microemulsion.
Further, in the water-soluble microemulsion, the surfactant is selected from: PPG-13-decyltetradecanol polyether-24.
Further, in the water-soluble microemulsion, the co-surfactant is selected from: PEG/PPG/polytetramethylene glycol-8/5/3 glycerol.
In another aspect, the present invention provides a method for preparing the above water-soluble microemulsion, which comprises the following steps: weighing the components: industrial hemp full spectrum oil, surfactant and cosurfactant for standby; adding industrial hemp full spectrum oil into ethanol, and stirring at high speed at 65 deg.C to dissolve completely; then adding a surfactant and a cosurfactant into the mixture at the temperature of 65 ℃, stirring the mixture at 600rpm until the mixture is uniform, and obtaining clear and transparent self-microemulsion; under the condition of natural cooling, the clear self-microemulsion is slowly dripped into ultrapure water, slowly stirred and fully dissolved to form the microemulsion.
Further, in the preparation method, the surfactant is selected from PPG-13-decyltetradecanol polyether-24, Tween 80, PEG40 hydrogenated castor oil.
Further, in the preparation method, the cosurfactant is selected from PEG/PPG/polytetramethylene glycol-8/5/3 glycerin, propylene glycol, isopropanol, polyethylene glycol and glycerin.
Further, in the preparation method, the surfactant is selected from: PPG-13-decyltetradecanol polyether-24.
Further, in the preparation method, the co-surfactant is selected from: PEG/PPG/polytetramethylene glycol-8/5/3 glycerol.
In another aspect, the present invention provides a water-soluble microemulsion prepared according to the preparation method.
In a further aspect, the present invention relates to the use of the above-described water-soluble microemulsions in the preparation of cosmetics.
In another aspect, the present invention relates to a cosmetic product comprising the above-described water-soluble microemulsion.
In another aspect, the present invention relates to the use of the above water-soluble microemulsion in the preparation of anti-allergic anti-inflammatory cosmetic products.
In another aspect, the present invention relates to the use of the above water-soluble microemulsion in anti-allergy and anti-inflammatory treatment of the skin.
In another aspect, the present invention relates to the use of the above water-soluble microemulsion for the preparation of an anti-allergic anti-inflammatory formulation.
In another aspect, the present invention relates to an anti-allergic anti-inflammatory formulation comprising the above-described water-soluble microemulsion.
In another aspect, the present invention relates to the use of the above water-soluble microemulsion for scavenging free radicals.
In another aspect, the present invention relates to the use of the above-described water-soluble microemulsion for inhibiting, ameliorating or alleviating infection by staphylococcus aureus and/or propionibacterium acnes on the skin.
In another aspect, the present invention relates to a formulation for inhibiting staphylococcus aureus and/or propionibacterium acnes, comprising the above-mentioned water-soluble microemulsion.
In another aspect, the present invention relates to the use of the above-described water-soluble microemulsion for the preparation of a formulation for inhibiting staphylococcus aureus and/or propionibacterium acnes.
In another aspect, the present invention relates to the use of the above water-soluble microemulsion for scavenging free radicals.
In another aspect, the present invention relates to a free radical scavenging formulation comprising the above-described water-soluble microemulsion.
In another aspect, the present invention relates to the use of the above water-soluble microemulsion for scavenging superoxide anions.
In another aspect, the present invention relates to a superoxide anion scavenging formulation comprising the above-described water-soluble microemulsion.
Advantageous effects
The water-soluble microemulsion provided by the invention has the advantages of simple preparation process and stable property, is easy to add into cosmetics of various formulations, and is suitable for industrial production. Meanwhile, the composition can effectively inhibit the activity of hyaluronidase, inhibit the secretion of IL-8 of human keratinocytes and promote the generation of IL-10, remarkably inhibit the growth of staphylococcus aureus and propionibacterium acnes, remove free radicals, and cannot generate obvious coagulation or hemolytic reaction, thereby relieving the inflammatory reaction of skin, realizing anti-inflammation and relieving, and having wide application prospect in anti-allergy and anti-inflammatory cosmetics.
Drawings
FIG. 1 is a transmission electron microscope image of water-soluble microemulsion, and the inset in the lower right corner is a real object photograph of the corresponding solution, as shown in the figure, the microemulsion has small particle size and uniform dispersion, and the emulsion is in a transparent state;
FIG. 2 is a distribution diagram of the particle size of the water-soluble microemulsion, which shows that the particle size of the water-soluble microemulsion is 10-30 nm;
FIG. 3 is a graphical representation of the inhibitory ability of water-soluble microemulsions on hyaluronidase activity;
FIG. 4 is a schematic illustration of the effect of water-soluble microemulsions on human keratinocyte IL-8 secretion;
FIG. 5 is a graph of the effect of water-soluble microemulsions on human keratinocyte IL-10 secretion;
FIG. 6 is a graph showing the effect of the water-soluble microemulsion on scavenging DPPH radicals;
FIG. 7 is a graph showing the results of the degree of hemolysis of a water-soluble microemulsion;
FIG. 8 is a graph showing the effect of water-soluble microemulsion on physiological morphology of erythrocytes.
Detailed Description
The invention is described in detail below with reference to the figures and the embodiments. The present invention includes, but is not limited to, the following embodiments.
Example 1 Process for preparing a Water-soluble microemulsion
The method specifically comprises the following steps: weighing the components: 1g of industrial hemp full-spectrum oil (RM 005 available from Yunnan Ci Ma-rui-hemp Biotech Co., Ltd.), 7g of PPG-13-decyltetradecyltetradecylpolyether-24, and 7g of PEG/PPG/polytetramethylene glycol-8/5/3 glycerol for standby; adding industrial hemp full spectrum oil into 0.4g ethanol, and stirring at high speed at 65 deg.C to dissolve completely; then adding PPG-13-decyltetradecanol polyether-24 and PEG/PPG/polytetramethylene glycol-8/5/3 glycerol at the temperature of 65 ℃, stirring at 600rpm until the mixture is uniform to obtain clear and transparent self-microemulsion; under the condition of natural cooling, the clear self-microemulsion is slowly dripped into ultrapure water, and the ultrapure water is slowly stirred and fully dissolved to form transparent microemulsion.
The transparent microemulsion prepared by the invention has the following characteristics: (1) in cosmetics, to make the presence of any non-homogeneous substances or precipitates easily detectable; (2) the microemulsion prepared by the invention is spontaneously formed and has the characteristics of energy conservation and high efficiency; (3) the microemulsion prepared by the invention can be stored for a long time without layering; (4) the microemulsion prepared by the invention does not need to add an oil phase; (5) the microemulsion prepared by the invention is uniformly distributed into droplets with the particle size of about 10-30 nm, and is beneficial to absorption of skin.
Example 2 detection of the inhibitory Capacity of Water-soluble microemulsions on Hyaluronidase Activity
The hyaluronidase inhibition experiment is the most typical in vitro method for evaluating the anti-allergic activity, the anti-allergic activity of the substance is evaluated by taking the hyaluronidase inhibition rate as an index,the greater the hyaluronidase inhibition, the greater the anti-allergic activity. In-vitro hyaluronidase inhibition experiments are carried out by an Elson-Morgan improvement method, and the anti-allergy effect of the water-soluble microemulsion is evaluated. The method comprises the following specific steps: 0.1mL of 2.5M CaCl was taken2Incubating the aqueous solution and 0.5mL hyaluronidase solution (500U/mL, with acetate buffer as solvent) at 37 deg.C for 20 min; adding 0.5mL of sample solution containing different concentrations of industrial hemp full spectrum oil (0.005 wt%, 0.01 wt%, 0.5 wt%, 1 wt%, 2 wt%), or 0.5mL of deionized water as positive control, and keeping the temperature at 37 deg.C for 20 min; adding 0.5mL sodium hyaluronate solution (0.5mg/mL, acetic acid buffer solution as solvent) at 37 deg.C for 30min, and standing at room temperature for 5 min; 0.1mL of 0.4mol/L NaOH solution and 0.5mL of acetylacetone solution (50mL of 1M NaCO) were added3Mixing the aqueous solution with 3.5mL of acetylacetone), heating in a boiling water bath for 15min, and immediately cooling with ice water for 5 min; adding 1.0mL of Ellisib reagent (0.8g p-dimethylaminobenzaldehyde dissolved in 15mL concentrated hydrochloric acid and 15mL absolute ethanol), standing for developing color for 20min, and measuring the absorbance at 535 nm. The results are shown in FIG. 3.
As can be seen from fig. 3, the inhibition rate of the water-soluble microemulsion of the present invention on hyaluronidase has a significant concentration dependency, and the inhibition rate of the water-soluble microemulsion obtained at a content of 1 wt% on hyaluronidase exceeds about 80%. When the concentration is increased to 2 wt%, the inhibition rate is increased to about 98.2%. Therefore, the obtained water-soluble microemulsion has better hyaluronidase inhibition capability, can maintain the cell barrier function to a certain extent, protects the skin from external stimulation and has the anti-allergy effect.
Example 3 Effect of Water-soluble microemulsions on human keratinocyte IL-8 and IL-10 secretion assay
HaCaT was treated with RPMI1640 containing 10% fetal bovine serum, 100U/ml penicillin, 100. mu.g/ml streptomycin at 37 ℃ with 5% CO2Culturing under saturated humidity condition. Selecting cells in logarithmic growth phase, inoculating to 96-well culture plate, culturing for 24 hr, replacing culture medium with RPMI1640 containing industrial hemp full-lineage oil test sample with different concentrations, incubating for 12 hr, collecting cell culture supernatant, and detecting with human IL-8 and IL-10ELISA kitThe concentrations of IL-8 and IL-10 in the clear liquid. The results are shown in FIGS. 4 and 5.
As can be seen from the figure, the water-soluble microemulsion with different industrial hemp full-lineage oil concentrations of the invention has the inhibition effect on the IL-8 secretion of human keratinocytes, and has the promotion effect on the IL-10 secretion. As the concentration increases, the inhibition of IL-8 becomes more obvious, and the secretion of IL-10 is in a descending trend.
EXAMPLE 4 Water-soluble microemulsion bacteriostasis test (Staphylococcus aureus, Propionibacterium acnes)
Vertically placing the oxford cup into a culture dish containing the bacterial liquid and the culture medium which are uniformly mixed, slightly pressing, filling the test sample aqueous solution into the cup, covering a cover of the culture dish, placing the culture dish at a proper temperature, culturing for 24 hours, and observing the existence and the size of a bacteriostatic circle around the oxford cup. The size of the inhibition zone reflects the sensitivity of the detection bacteria to the determination drug, and is in negative correlation with the Minimum Inhibitory Concentration (MIC) of the drug to the bacteria to be determined, namely, the larger the inhibition zone is, the smaller the MIC is. The results are shown in Table 1.
TABLE 1 antibacterial zone size of Staphylococcus aureus and Propionibacterium acnes
| Full spectrum oil water soluble microemulsion | Diameter of inhibition zone (Propionibacterium acnes) |
| 2wt％ | 25.4mm |
| 1wt％ | 20mm |
| 0.5wt％ | 20mm |
| 0.1wt％ | 14.8mm |
| 0.05wt％ | 12.6mm |
| 0.005wt％ | 7.3mm |
| H2O | 7.4mm |
The results show that the diameter of the inhibition zone of the matrix water is 7.4mm, and the water-soluble microemulsion of the invention has the inhibition effect on staphylococcus aureus and propionibacterium acnes at 0.05 wt%.
EXAMPLE 5 Water-soluble microemulsion free radical DPPH scavenging experiment
Taking 0.003g of DPPH, using absolute ethyl alcohol to fix the volume to 50mL, using the absorption peak of characteristic purple-red color mass of DPPH solution at 517nm, using ultraviolet-visible spectrophotometry to determine the absorbance reduction after adding the water-soluble microemulsion sample (the concentration of industrial hemp full-spectrum oil is 0.01 wt%) or vitamin C sample (VC) to represent and compare the free radical scavenging capacity of the water-soluble microemulsion of the invention, and the following table adds reaction liquid in groups.
The DPPH assay was loaded as in table 2.
TABLE 2 DPPH method experiment sample adding table
| Sample name | Sample reaction solution |
| A sample | 1mL sample +1mL DPPH solution |
| A control | 1mL of absolute ethanol +1mL of DPPH solution |
| A sample blank | 1mL of absolute ethanol and 1mL of deionized water |
After shaking vigorously, the sample represented by a sample was allowed to stand at room temperature for 30min, and then added to a cuvette to measure absorbance at 517nm, absorbance values represented by a sample, a control and a sample blank were measured, and the clearance SA (%) was calculated as 1- (a sample + a sample blank)/a control × 100, a control — absorbance of DPPH solution without sample addition; sample-absorbance of DPPH solution after addition of sample; a sample blank is the absorbance of the mixture of deionized water and absolute ethanol.
The results are shown in fig. 6, and the experimental results show that the speed of removing DPPH free radicals by the water-soluble microemulsion is higher than that of VC, the antioxidant effect is good, and the water-soluble microemulsion has a strong effect of removing DPPH free radicals and is helpful for relieving skin.
EXAMPLE 6 superoxide anion scavenging experiment with water-soluble microemulsion
The pyrogallol autoxidation reaction generates intermediate products and superoxide anions, the intermediate products and the superoxide anions have obvious ultraviolet absorption peaks at specific wavelengths, and the scavenging capacity of the intermediate products and the superoxide anions is shown by measuring the reduction of absorbance after adding water-soluble microemulsion by using an ultraviolet-visible spectrophotometry. The following table groups the reaction solution.
4.5mL of 0.05mol/L Tris-HCl buffer solution (pH8.2) and 0.4mL of 25mmol/L pyrogallol solution (2.5mmol/L pyrogallol (prepared by 10mmol/L HCl)) were mixed well, and then reacted in a water bath at 25 ℃ for 5 min.
TABLE 3 sample adding table for experiment of superoxide anion method
After shaking vigorously, the mixture was added to a cuvette to measure absorbance at 299nm, and absorbance values indicated by A sample and A control were measured. The results are shown in Table 4.
TABLE 4 absorbance values at 299nm of superoxide anion process
| Time/min | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| A sample/A | 0.23 | 0.381 | 0.501 | 0.62 | 0.721 | 0.817 | 0.895 | 0.975 | 1.113 | 1.201 |
| A control/A | 0.15 | 0.303 | 0.441 | 0.553 | 0.654 | 0.741 | 0.819 | 0.895 | 1.025 | 1.059 |
The experimental result shows that the water-soluble microemulsion can remove superoxide anions along with time, has good antioxidant effect and is beneficial to relieving skin.
EXAMPLE 6 hemolysis assay of water-soluble microemulsions
The particle size of the water-soluble microemulsion is 10-30 nm, so that the water-soluble microemulsion is beneficial to skin permeation and is likely to enter blood. Therefore, the potential hemolysis thereof was evaluated and the blood safety of the water-soluble microemulsion was investigated. In vivo hemolysis (red blood cell rupture) can lead to anemia, jaundice, and other pathological conditions.
According to the principle that heme released by erythrocyte rupture has maximum absorption in a visible light wavelength range, the hemolytic degree of the test solution is detected by adopting a spectrophotometry method.
A4% suspension of red blood cells (v/v, 0.5mL) was incubated with the sample solution at 37 ℃ for 6 hours. The supernatant from the low-speed centrifugation was collected and the absorbance at 562nm was measured. Here, 4% erythrocyte suspension (v/v, 0.5mL) was incubated with deionized water and the centrifuged supernatant was used as a positive control. Calculating Hemolysis ratio (A/A0 × 100), A-absorbance of red blood cell solution added with deionized water; a0-absorbance of the erythrocyte solution after addition of the sample solution. The results are shown in FIGS. 7 and 8.
As can be seen from the figure, the hemolysis rate in the measured concentration range does not exceed 5%, and the blood safety is good. Meanwhile, the physiological morphology of the red blood cells is hardly influenced.
The above examples of the present invention are merely examples for clearly illustrating the present invention and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the claims of the present invention.
Claims (10)
1. A water-soluble microemulsion characterized by comprising cannabidiol, a surfactant, a co-surfactant and water.
2. The water soluble microemulsion according to claim 1 characterized in that said cannabidiol is selected from the group consisting of: the industrial hemp full-spectrum oil contains cannabidiol, and the weight percentage is 45-55 wt%; optionally, the surfactant is selected from PPG-13-decyltetradecanol polyether-24, tween 80, PEG40 hydrogenated castor oil; optionally, the co-surfactant is selected from PEG/PPG/polytetramethylene glycol-8/5/3 glycerol, propylene glycol, isopropanol, polyethylene glycol, glycerol.
3. The water-soluble microemulsion according to claim 2, characterized in that the content in percentage by weight of the industrial cannabis full spectrum oil is: 0.5 to 5 wt%, preferably 0.5 to 2 wt%, and more preferably 0.5 to 1 wt% of the total weight of the water-soluble microemulsion; optionally, the weight percentage of the surfactant is: 2-40 wt%, preferably 2-30 wt%, more preferably 2-7 wt% of the total weight of the water-soluble microemulsion; optionally, the weight percentage of the co-surfactant is: 2-40 wt%, preferably 2-30 wt%, more preferably 2-7 wt% of the total weight of the water-soluble microemulsion.
4. The water-soluble microemulsion according to claim 2 or 3, characterized in that said surfactant is selected from: PPG-13-decyltetradecanol polyether-24; optionally, the co-surfactant is selected from: PEG/PPG/polytetramethylene glycol-8/5/3 glycerol.
5. Cosmetic product comprising a water-soluble microemulsion according to any one of claims 1 to 4.
6. The method of preparing a water-soluble microemulsion according to any one of claims 1 to 4, characterized by comprising the steps of: weighing the components: industrial hemp full spectrum oil, surfactant and cosurfactant for standby; adding industrial hemp full spectrum oil into ethanol, and stirring at high speed at 65 deg.C to dissolve completely; then adding a surfactant and a cosurfactant into the mixture at the temperature of 65 ℃, stirring the mixture at 600rpm until the mixture is uniform, and obtaining clear and transparent self-microemulsion; under the condition of natural cooling, the clear self-microemulsion is slowly dripped into ultrapure water, and the ultrapure water is slowly stirred and fully dissolved to form transparent microemulsion.
7. Anti-allergic anti-inflammatory preparation, characterized in that it comprises a water-soluble microemulsion according to any one of claims 1 to 4.
8. Preparation inhibiting staphylococcus aureus and/or propionibacterium acnes, characterized in that it comprises a water-soluble microemulsion according to any one of claims 1 to 4.
9. Formulation for scavenging free radicals or for scavenging superoxide anions, characterized in that it comprises a water-soluble microemulsion according to any one of claims 1 to 4.
10. Use of a water-soluble microemulsion according to any one of claims 1 to 4 for the preparation of a cosmetic, an anti-allergic, anti-inflammatory preparation, a preparation inhibiting staphylococcus aureus and/or propionibacterium acnes, a preparation scavenging free radicals or a preparation scavenging superoxide anions.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110013004.3A CN112675077A (en) | 2021-01-06 | 2021-01-06 | Water-soluble microemulsion and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110013004.3A CN112675077A (en) | 2021-01-06 | 2021-01-06 | Water-soluble microemulsion and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112675077A true CN112675077A (en) | 2021-04-20 |
Family
ID=75456006
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110013004.3A Pending CN112675077A (en) | 2021-01-06 | 2021-01-06 | Water-soluble microemulsion and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112675077A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113616683A (en) * | 2021-09-16 | 2021-11-09 | 中国农业科学院麻类研究所 | Apocynum venetum flavone self-microemulsion method and application thereof |
| CN114712274A (en) * | 2021-12-23 | 2022-07-08 | 上海交通大学医学院附属第九人民医院 | Soothing and anti-allergy nano vesicle composition and application thereof |
| CN116211742A (en) * | 2022-12-28 | 2023-06-06 | 上海林清轩生物科技有限公司 | Microemulsion and preparation method thereof |
-
2021
- 2021-01-06 CN CN202110013004.3A patent/CN112675077A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113616683A (en) * | 2021-09-16 | 2021-11-09 | 中国农业科学院麻类研究所 | Apocynum venetum flavone self-microemulsion method and application thereof |
| CN114712274A (en) * | 2021-12-23 | 2022-07-08 | 上海交通大学医学院附属第九人民医院 | Soothing and anti-allergy nano vesicle composition and application thereof |
| CN114712274B (en) * | 2021-12-23 | 2023-05-16 | 上海康华时代生物医学科技有限公司 | Nanometer vesicle composition for relieving and resisting allergy and application thereof |
| CN116211742A (en) * | 2022-12-28 | 2023-06-06 | 上海林清轩生物科技有限公司 | Microemulsion and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112675077A (en) | Water-soluble microemulsion and preparation method thereof | |
| CN108324606B (en) | Pitaya stem extract and preparation method and application thereof | |
| CN111557875B (en) | Skin care composition for resisting light pollution and light injury and preparation method and application thereof | |
| CN103536455B (en) | Application of compound bacteria polysaccharide in preparing moisture-keeping cosmetic | |
| CN107320375A (en) | Bioactivity vegetalization cosmetic compositions and its production method | |
| KR100799991B1 (en) | Cosmetic composition for wrinkle care | |
| CN113633562B (en) | Matrine vegetable oil eutectic solvent and preparation method, application and product thereof | |
| CN111407678A (en) | Microemulsion with moisturizing, skin-brightening and anti-aging effects and preparation method thereof | |
| CN111657415B (en) | Curcumin nanoparticle and preparation method and application thereof | |
| Zhao et al. | Chitosan-thymol nanoparticle with pH responsiveness as a potential intelligent botanical fungicide against Botrytis cinerea | |
| CN113413404A (en) | Traditional Chinese medicine preparation with anti-inflammation and acne-removing effects and preparation method and application thereof | |
| KR102654331B1 (en) | Preparation and application of a fermentation composition with anti-photoaging,calming,cooling effects | |
| CN112137937B (en) | Active composition with skin aging delaying function and application thereof | |
| CN110638738B (en) | Method for preparing ecological nutrient solution by treating roses in low-temperature bath and application | |
| CN112402286A (en) | Sunscreen cream and preparation method thereof | |
| CN111840162A (en) | Tea polyphenol and ethanol double-disinfection and sterilization wash-free hand sanitizer and preparation method thereof | |
| CN117883352B (en) | Compact anti-wrinkle composition and application thereof | |
| CN110693795A (en) | Skin care product containing purple rice anthocyanin and preparation method thereof | |
| CN103622875A (en) | Preparation method of catechin hydrogel skin care product | |
| CN104127406A (en) | Application of dihydromyricetin in preparing inhibitor for liver cell oxidative damage | |
| CN112716844B (en) | Hand cream and preparation method thereof | |
| CN104606115B (en) | Rose honey and its application | |
| CN118178271B (en) | Composition for improving skin state and application thereof | |
| CN112137935B (en) | Composition and application thereof | |
| CN115813838B (en) | Cosmetic with anti-aging composition and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210420 |
|
| RJ01 | Rejection of invention patent application after publication |