CN114280200B - Method for simultaneously detecting three component contents of praziquantel, clofenacin and milbexime - Google Patents
Method for simultaneously detecting three component contents of praziquantel, clofenacin and milbexime Download PDFInfo
- Publication number
- CN114280200B CN114280200B CN202111657868.5A CN202111657868A CN114280200B CN 114280200 B CN114280200 B CN 114280200B CN 202111657868 A CN202111657868 A CN 202111657868A CN 114280200 B CN114280200 B CN 114280200B
- Authority
- CN
- China
- Prior art keywords
- praziquantel
- impurity
- milbexime
- clofenacin
- solution
- 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.)
- Active
Links
- 229960002957 praziquantel Drugs 0.000 title claims abstract description 50
- FSVJFNAIGNNGKK-UHFFFAOYSA-N 2-[cyclohexyl(oxo)methyl]-3,6,7,11b-tetrahydro-1H-pyrazino[2,1-a]isoquinolin-4-one Chemical compound C1C(C2=CC=CC=C2CC2)N2C(=O)CN1C(=O)C1CCCCC1 FSVJFNAIGNNGKK-UHFFFAOYSA-N 0.000 title claims abstract description 49
- 238000000034 method Methods 0.000 title claims abstract description 36
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 37
- 239000003085 diluting agent Substances 0.000 claims abstract description 23
- 239000000523 sample Substances 0.000 claims abstract description 21
- 239000000243 solution Substances 0.000 claims abstract description 21
- 239000012488 sample solution Substances 0.000 claims abstract description 17
- 239000007788 liquid Substances 0.000 claims abstract description 16
- 239000012490 blank solution Substances 0.000 claims abstract description 14
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000012085 test solution Substances 0.000 claims abstract description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000002347 injection Methods 0.000 claims abstract description 7
- 239000007924 injection Substances 0.000 claims abstract description 7
- 238000000825 ultraviolet detection Methods 0.000 claims abstract description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims abstract description 4
- 238000010828 elution Methods 0.000 claims abstract description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000012535 impurity Substances 0.000 claims description 29
- 239000013558 reference substance Substances 0.000 claims description 14
- RPNVVNYKJLHAFP-UHFFFAOYSA-N n-formyl-n-[2-oxo-2-(1-oxo-3,4-dihydroisoquinolin-2-yl)ethyl]cyclohexanecarboxamide Chemical compound C1CC2=CC=CC=C2C(=O)N1C(=O)CN(C=O)C(=O)C1CCCCC1 RPNVVNYKJLHAFP-UHFFFAOYSA-N 0.000 claims description 10
- 238000005303 weighing Methods 0.000 claims description 9
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 claims description 7
- RAZNCTHEBQFYML-UHFFFAOYSA-N 2-(cyclohexanecarbonyl)-6,7-dihydro-3h-pyrazino[2,1-a]isoquinolin-4-one Chemical compound C1C(=O)N2CCC3=CC=CC=C3C2=CN1C(=O)C1CCCCC1 RAZNCTHEBQFYML-UHFFFAOYSA-N 0.000 claims description 7
- XEYCCEDRIDKXEV-UHFFFAOYSA-N 2-benzoyl-3,6,7,11b-tetrahydro-1h-pyrazino[2,1-a]isoquinolin-4-one Chemical compound C1C(C2=CC=CC=C2CC2)N2C(=O)CN1C(=O)C1=CC=CC=C1 XEYCCEDRIDKXEV-UHFFFAOYSA-N 0.000 claims description 7
- AAEQXEDPVFIFDK-UHFFFAOYSA-N 3-(4-fluorobenzoyl)-2-(2-methylpropanoyl)-n,3-diphenyloxirane-2-carboxamide Chemical compound C=1C=CC=CC=1NC(=O)C1(C(=O)C(C)C)OC1(C=1C=CC=CC=1)C(=O)C1=CC=C(F)C=C1 AAEQXEDPVFIFDK-UHFFFAOYSA-N 0.000 claims description 7
- 239000011259 mixed solution Substances 0.000 claims description 6
- -1 2, 6-difluorobenzoyl Chemical group 0.000 claims description 5
- PVKCAQKXTLCSBC-UHFFFAOYSA-N 1h-isoquinolin-4-one Chemical compound C1=CC=C2C(=O)C=NCC2=C1 PVKCAQKXTLCSBC-UHFFFAOYSA-N 0.000 claims description 3
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 claims description 3
- 125000001951 carbamoylamino group Chemical group C(N)(=O)N* 0.000 claims description 3
- ROORDVPLFPIABK-UHFFFAOYSA-N diphenyl carbonate Chemical compound C=1C=CC=CC=1OC(=O)OC1=CC=CC=C1 ROORDVPLFPIABK-UHFFFAOYSA-N 0.000 claims description 3
- 125000005646 oximino group Chemical group 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims 1
- 229910002027 silica gel Inorganic materials 0.000 claims 1
- 238000001514 detection method Methods 0.000 abstract description 16
- 239000003814 drug Substances 0.000 abstract description 5
- 239000000377 silicon dioxide Substances 0.000 abstract description 3
- 238000012360 testing method Methods 0.000 description 14
- KPYSYYIEGFHWSV-UHFFFAOYSA-N Baclofen Chemical compound OC(=O)CC(CN)C1=CC=C(Cl)C=C1 KPYSYYIEGFHWSV-UHFFFAOYSA-N 0.000 description 12
- 238000011084 recovery Methods 0.000 description 11
- 230000015556 catabolic process Effects 0.000 description 8
- 238000006731 degradation reaction Methods 0.000 description 8
- 239000011550 stock solution Substances 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- 239000000047 product Substances 0.000 description 7
- 241000238631 Hexapoda Species 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000007865 diluting Methods 0.000 description 6
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 244000045947 parasite Species 0.000 description 5
- 239000000575 pesticide Substances 0.000 description 5
- 238000012795 verification Methods 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 description 3
- HRYILSDLIGTCOP-UHFFFAOYSA-N N-benzoylurea Chemical compound NC(=O)NC(=O)C1=CC=CC=C1 HRYILSDLIGTCOP-UHFFFAOYSA-N 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 3
- 239000000077 insect repellent Substances 0.000 description 3
- 230000000749 insecticidal effect Effects 0.000 description 3
- CKVMAPHTVCTEMM-ALPQRHTBSA-N milbemycin oxime Chemical compound O1[C@H](C)[C@@H](C)CC[C@@]11O[C@H](C\C=C(C)\C[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)C(=N/O)/[C@H]3OC\2)O)C[C@H]4C1.C1C[C@H](C)[C@@H](CC)O[C@@]21O[C@H](C\C=C(C)\C[C@@H](C)\C=C\C=C/1[C@]3([C@H](C(=O)O4)C=C(C)C(=N/O)/[C@H]3OC\1)O)C[C@H]4C2 CKVMAPHTVCTEMM-ALPQRHTBSA-N 0.000 description 3
- 229940099245 milbemycin oxime Drugs 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 239000012086 standard solution Substances 0.000 description 3
- 241000242722 Cestoda Species 0.000 description 2
- 241000869417 Trematodes Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000000890 antigenic effect Effects 0.000 description 2
- 239000003096 antiparasitic agent Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000004128 high performance liquid chromatography Methods 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- 239000003120 macrolide antibiotic agent Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- ZLBGSRMUSVULIE-GSMJGMFJSA-N milbemycin A3 Chemical compound O1[C@H](C)[C@@H](C)CC[C@@]11O[C@H](C\C=C(C)\C[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 ZLBGSRMUSVULIE-GSMJGMFJSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000012088 reference solution Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- 229920002101 Chitin Polymers 0.000 description 1
- 208000003917 Dirofilariasis Diseases 0.000 description 1
- 241000242711 Fasciola hepatica Species 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 101000777134 Homo sapiens Ubiquitin carboxyl-terminal hydrolase 43 Proteins 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 239000005912 Lufenuron Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- BWCRYQGQPDBOAU-UHFFFAOYSA-N Milbemycin D Natural products C1CC(C)C(C(C)C)OC21OC(CC=C(C)CC(C)C=CC=C1C3(C(C(=O)O4)C=C(C)C(O)C3OC1)O)CC4C2 BWCRYQGQPDBOAU-UHFFFAOYSA-N 0.000 description 1
- 206010062575 Muscle contracture Diseases 0.000 description 1
- 102100031311 Ubiquitin carboxyl-terminal hydrolase 43 Human genes 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 208000006111 contracture Diseases 0.000 description 1
- SRONXYPFSAKOGH-UHFFFAOYSA-N cyclopentadecane Chemical compound C1CCCCCCCCCCCCCC1 SRONXYPFSAKOGH-UHFFFAOYSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000002865 immune cell Anatomy 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 239000002949 juvenile hormone Substances 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 238000012417 linear regression Methods 0.000 description 1
- PWPJGUXAGUPAHP-UHFFFAOYSA-N lufenuron Chemical compound C1=C(Cl)C(OC(F)(F)C(C(F)(F)F)F)=CC(Cl)=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F PWPJGUXAGUPAHP-UHFFFAOYSA-N 0.000 description 1
- 229960000521 lufenuron Drugs 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- FXWHFKOXMBTCMP-WMEDONTMSA-N milbemycin Natural products COC1C2OCC3=C/C=C/C(C)CC(=CCC4CC(CC5(O4)OC(C)C(C)C(OC(=O)C(C)CC(C)C)C5O)OC(=O)C(C=C1C)C23O)C FXWHFKOXMBTCMP-WMEDONTMSA-N 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 230000017448 oviposition Effects 0.000 description 1
- 238000010525 oxidative degradation reaction Methods 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000037081 physical activity Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 125000002572 propoxy group Chemical group [*]OC([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 1
- LMKOGFPNETZZTE-UHFFFAOYSA-N pyrido[2,3-f]quinoxaline Chemical class C1=CN=C2C3=NC=CC=C3C=CC2=N1 LMKOGFPNETZZTE-UHFFFAOYSA-N 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000011003 system suitability test Methods 0.000 description 1
- 239000012489 system suitability test solution Substances 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 239000000273 veterinary drug Substances 0.000 description 1
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a method for simultaneously detecting the contents of praziquantel, clofenacin and milbexime, which comprises the steps of preparing a blank solution, preparing a sample solution containing praziquantel, clofenacin and milbexime, preparing a system applicability test solution, respectively injecting the solution into a liquid chromatograph, and recording a chromatogram under ultraviolet wavelength; the diluent is acetonitrile, octadecylsilane chemically bonded silica is used as a chromatographic column in a liquid chromatograph, the gradient elution is carried out, the flow rate is 1.0mL/min, the column temperature is 30 ℃, the ultraviolet detection wavelength is 230nm, and the sample injection amount is 20 mu L; the mobile phase A is phosphoric acid solution with the content of 10-30wt% and the mobile phase B is acetonitrile with the content of 70-90wt%; the detection method is simple and convenient, saves time, reduces cost, has good specificity, accuracy, linearity, durability and precision, is suitable and accurate for detecting the content of the three components, and can be used for daily detection of the pet medicine.
Description
Technical Field
The invention belongs to the technical field of chemical analysis, and particularly relates to a method for simultaneously detecting the contents of praziquantel, clofenacet and milbexime.
Background
Praziquantel is a pyrazinoquinoline derivative with insect repellent activity, developed by Bayer company in the 70 th century, and is a broad-spectrum anti-fluke and tapeworm drug.
The mechanism of action of praziquantel is not fully elucidatedMainstream theory suggests that praziquantel may alter the body to Ca 2+ The permeability of the artificial limb promotes the inflow of the artificial limb to cause the excitation of the insect body activity and the contracture of muscles, so that the artificial limb cannot be well adsorbed on the wall of a blood vessel; on the other hand, the damage to the cortex of the insect body also exposes the antigenic determinants on the surface of the insect body, so that the antigenic determinants are recognized by the immune system of the host and attract immune cells to attack the same. Thus, the insecticidal mechanism of praziquantel can be summarized as two parts, namely the direct action of the drug on the insect body and the immune effect of the host.
The clofenacin is a benzoyl urea pesticide, and is the first large product in the benzoyl urea pesticide. The new major market for clofen is in the field of animal health and is used for controlling fleas on cats and dogs.
Such pesticides are insect growth regulators that inhibit further growth of insect exoskeletons by inhibiting chitin synthesis. The product has no systemic property and stomach toxicity. While the peeling off accompanies the entire life cycle of the insect, benzoylurea insecticides are most effective against the larval molting stage. The product has poor quick-acting property, and is usually compounded with other traditional pesticides to improve the quick-acting property and the lasting effect of the product.
Milbemycin oxime is semisynthetic macrolide antiparasitic drug, and is milbemycin A 3 And milbemycins A 4 Is developed jointly by Sankyo and Ciba-Geigy (now nohua), and is initially used clinically for the control of heartworm disease and other endoparasitic infections.
The mechanism of action of milbexime is similar to that of other macrolide antiparasitic drugs, milbexime acts as an agonist of GABA (gamma-aminobutyric acid) neurotransmitter in nerve cells, and can also act as an invertebrateGlutamate-gated Cl in neural and muscle cells of the species – The channels are combined. In both cases, nerve signaling from the parasite is blocked, paralyzing the parasite, and causing death of the parasite. Milbexime also affects the reproduction of some parasites by reducing the oviposition of the parasite or inducing the production of abnormal eggs. However, due to lack of GABA-mediated mediators and glutamate-gated Cl in cestodes and trematodes – Channels, milbexime is not effective against tapeworms and trematodes.
The insecticidal spectrum of the above 3 components is complementary, and the combination application of the insecticidal composition to the pet insect repellent has very good and broad-spectrum effect. At present, the oral pet insect repellent prepared by the 3 components is already present on overseas markets, but the content detection method is not disclosed. In the presently disclosed data, these three components need to be detected by three different HPLC methods.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide a method for simultaneously detecting the contents of praziquantel, clofenacet and milbeoxime.
To achieve the above object, the solution of the present invention is:
a method for simultaneously detecting the contents of praziquantel, clofenacet and milbexime, which comprises the following steps:
(1) Taking the diluent as a blank solution, injecting the blank solution into a liquid chromatograph, and recording chromatograms at wavelengths of 210nm, 230nm and 255 nm;
(2) Respectively weighing three components of praziquantel, clofenacin and milbexime, adding a diluent to prepare a solution containing 26 mug of praziquantel, 73 mug of clofenacin and 4.8 mug of milbexime as sample solutions per 1mL, injecting the sample solutions into a liquid chromatograph, and recording chromatograms at wavelengths of 210nm, 230nm and 255 nm;
(3) And taking a proper amount of each of praziquantel, praziquantel impurity A, praziquantel impurity B, praziquantel impurity C, clofenacin impurity B, clofenacin impurity C, clofenacin impurity G, milbexime and milbexime impurity D, adding a diluent to dissolve and prepare a mixed solution containing 1mg of each 1mL of each of the above-mentioned reference substances as a system applicability test solution, injecting the mixed solution into a liquid chromatograph, and recording chromatograms at wavelengths of 230nm and 255 nm.
Preferably, in step (2), praziquantel is present in a concentration of 283.20-654.64. Mu.g/mL, clofenacin is present in a concentration of 570.44-1171.20. Mu.g/mL, milbexime is present in a concentration of 27.57-64.32. Mu.g/mL.
Preferably, in step (3), praziquantel impurity A is (11 bRS) -2-benzoyl-1, 2,3,6,7, 11B-hexahydro-4H-pyrazinyl [2,1-a ] isoquinolin-4-one, praziquantel impurity B is 2- (cyclohexaformyl) -2,3,6, 7-tetrahydro-4H-pyrazino [2,1-a ] isoquinolin-4-one, and praziquantel impurity C is N-formyl-N- [ 2-oxo-2- (1-oxo-3, 4-dihydroisoquinolin-2 (1H) -yl) ethyl ] cyclohexanecarboxamide.
Preferably, in step (3), the novel impurity B of clofenacin is N- [ (2, 5-dichloro-4-p-hydroxybenzoyl) carbamoyl ] -2, 6-difluorobenzamide, the novel impurity C of clofenacin is N- [ 3-chloro-4- (1, 2, 3-perfluorobutadienropoxy) phenyl-carbamoyl ] -2, 6-difluorobenzamide, and the novel impurity G of clofenacin is 2, 5-dichloro-4- [3- (2, 6-difluorobenzoyl) ureido ] diphenyl carbonate.
Preferably, in step (3), milbeoxime impurity D is (1 ' r,2r,4's,5s,6r,8' r,10' e,13' r,14' e,16' e,20' r,21' z, 24's) -24' -hydroxy-21 ' - (oximino) -6- (propan-2-yl) -5,11',13',22' tetramethyl-3, 4,5, 6-tetrahydrospirocyclo [ pyran-2, 6' - [3,7,19] trioxabicyclo [15.6.1.14,8.020,24] cyclopentadec [10,14,16,22] tetraen ] -2' -one.
Preferably, in step (1), step (2) and step (3), the diluent is acetonitrile.
Preferably, in step (1), step (2) and step (3), octadecylsilane chemically bonded silica is used as filler NUCLEOSIL C18 column (4.6X108 mm,5 μm) in liquid chromatograph.
Preferably, in the step (1), the step (2) and the step (3), gradient elution is carried out in a liquid chromatograph, the flow rate is 1.0mL/min, the column temperature is 30 ℃, the ultraviolet detection wavelength is 230nm, and the sample injection amount is 20 mu L; the mobile phase A is 0.01% (v/v) phosphoric acid solution, the mobile phase B is acetonitrile, the content of the mobile phase A is 10-30wt%, and the content of the mobile phase B is 70-90wt%.
By adopting the scheme, the invention has the beneficial effects that:
the detection method is simple and convenient, saves time, reduces cost, has good specificity, accuracy, linearity, durability and precision, is suitable and accurate for detecting the content of the three components, and can be used for daily detection of the pet medicine.
Drawings
FIG. 1 is a blank solution chromatogram at 210nm according to the present invention.
FIG. 2 is a 230nm blank solution chromatogram of the present invention.
FIG. 3 is a 255nm blank solution chromatogram of the present invention.
FIG. 4 is a chromatogram of a 210nm sample solution of the present invention.
FIG. 5 is a chromatogram of a 230nm sample solution of the present invention.
FIG. 6 is a chromatogram of a 255nm sample solution of the present invention.
FIG. 7 is a chromatogram of a 230nm system applicability test solution of the present invention.
FIG. 8 is a chromatogram of a 255nm system suitability test solution of the present invention.
Fig. 9 is a graph of average peak area versus concentration for praziquantel in example 3 of the present invention.
FIG. 10 is a graph of the average peak area versus concentration of clofen in example 3 of the invention.
FIG. 11 is a plot of average peak area versus concentration for milbeoxime in example 3 of the present invention.
Detailed Description
The invention provides a method for simultaneously detecting the contents of praziquantel, clofenacet and milbexime.
From the literature it is known that: praziquantel (Praziquantel) ultraviolet detection wavelength of 210nm [1] The ultraviolet detection wavelength of the clofenacin (Lufenuron) is 255nm [2] Milbeoxime (Milbemycin Oxime) ultraviolet detection wavelength of 230nm [3] 。
The method for simultaneously detecting the contents of praziquantel, clofenacet and milbexime comprises the following steps:
(1) The diluent acetonitrile was used as a blank solution, 20. Mu.L of the blank solution was precisely measured, and the blank solution was injected into a liquid chromatograph to record chromatograms at 3 ultraviolet wavelengths (210 nm, 230nm and 255 nm), as shown in FIGS. 1 to 3.
(2) The three components were weighed and acetonitrile was added to prepare about 26. Mu.g of praziquantel, about 73. Mu.g of clofenacin and about 4.8. Mu.g of milbexime as test solutions per 1mL of praziquantel, 20. Mu.L of the solution was precisely measured, and the solution was injected into a liquid chromatograph to record chromatograms at 3 ultraviolet wavelengths (210 nm, 230nm and 255 nm), as shown in FIGS. 4 to 6 and tables 1 to 3.
(3) And taking a proper amount of each of the reference substances of praziquantel, praziquantel impurity A, praziquantel impurity B, praziquantel impurity C, clofenacin impurity B, clofenacin impurity C, clofenacin impurity G, milbexime and milbexime impurity D, adding acetonitrile to dissolve and dilute the mixture to prepare a mixed solution of which each 1mL contains about 1mg of the reference substances, and taking the mixed solution as a system applicability test solution. 20. Mu.L was precisely measured, and the solution was injected into a liquid chromatograph to record chromatograms at ultraviolet wavelengths of 230nm and 255nm, as shown in FIGS. 7 and 8, and tables 4 and 5.
As can be seen from fig. 1 to 3, the blank solution-acetonitrile peak did not interfere with each component peak (component peaks all appeared after 5 minutes).
As can be seen from fig. 4 to 6, the peak areas of praziquantel and clofen at 210nm are relatively maximum, but 210nm is the terminal absorption for milbexime and the peak area is small, so that the praziquantel and clofen are not selected; the peak area of praziquantel at 255nm is the smallest, and the peak areas of other components at 230nm and 255nm are not greatly different, so that 230nm is selected as the detection wavelength.
As can be seen from fig. 7 to 8, comparing the two wavelength detection results, the separation degree between each component peak and the front and rear impurity peaks in the system applicability test solution is greater than 2.0 at the wavelength of 230nm, and the theoretical plate number of each component peak is greater than 5000. Thus, a detection wavelength of 230nm was used for the three-component content measurement.
TABLE 1
Description DAD1A，Sig＝210，4Ref＝off
TABLE 2
Description DAD1B，Sig＝230，4Ref＝off
TABLE 3 Table 3
Description DAD1C，Sig＝255，4Ref＝off
TABLE 4 Table 4
Description DAD1B，Sig＝230，4Ref＝off
TABLE 5
Description DAD1C，Sig＝255，4Ref＝off
Under the chromatographic condition, four main component peaks-praziquantel, clofenacin and milbexime A 3 Milbeoxime A 4 Each impurity had no interference to the four major component peaks, and the repeatability test praziquantel RSD was 0.38% (n=6), the clofenacin RSD was 0.42% (n=6), and the milbexime RSD was 0.26% (n=6), which can be completely separated.
Wherein,,
1) According to European pharmacopoeia EP9.0 [1] Known impurities of praziquantel are known: praziquantel impurity A, praziquantel impurity B and praziquantel impurity C.
Praziquantel impurity A: (11 bRS) -2-benzoyl-1, 2,3,6,7,11 b-hexahydro-4H-pyrazinyl [2,1-a ] isoquinolin-4-one is a synthetic impurity generated during the production process.
Praziquantel impurity B:2- (cyclohexanecarbonyl) -2,3,6, 7-tetrahydro-4H-pyrazino [2,1-a ] isoquinolin-4-one is a synthetic impurity produced in the production process.
Praziquantel impurity C: N-formyl-N- [ 2-oxo-2- (1-oxo-3, 4-dihydroisoquinolin-2 (1H) -yl) ethyl ] cyclohexanecarboxamide is a degradation impurity of praziquantel.
2) According to the United states pharmacopoeia USP43 [2] Known impurities of clofenacin are: clofenacin impurity B, clofenacin impurity C and clofenacin impurity G.
Clofenacin impurity B: n- [ (2, 5-dichloro-4-p-hydroxybenzoyl) carbamoyl ] -2, 6-difluorobenzamide is a synthetic impurity produced in the production process.
Clofenacin impurity C: n- [ 3-chloro-4- (1, 2, 3-perfluorobutadiene propoxy) phenyl-carbamoyl ] -2, 6-difluorobenzamide is a synthetic impurity produced in the production process.
Clofenacin impurity G:2, 5-dichloro-4- [3- (2, 6-difluorobenzoyl) ureido ] diphenyl carbonate is a synthetic impurity produced in the production process.
3) According to the literature [3] Known impurities of milbeoxime are known: milbeoxime impurity D.
Milbeoxime impurity D: (1 ' R,2R,4' S,5S,6R,8' R,10' E,13' R,14' E,16' E,20' R,21' Z,24' S) -24' -hydroxy-21 ' - (oximino) -6- (propan-2-yl) -5,11',13',22' tetramethyl-3, 4,5, 6-tetrahydrospirocyclo [ pyran-2, 6' - [3,7,19] trioxytetracyclo [15.6.1.14,8.020,24] cyclopentadecane [10,14,16,22] tetraen ] -2' -one (milbemycin D oxime) is a synthetic impurity produced during production.
The specific verification process is as follows:
description of the detection method
Chromatographic column: octadecylsilane chemically bonded silica is used as filler (NUCLEOSIL C18X 4.6mm,5 μm or column with comparable performance).
Flow rate: 1.0mL/min.
Column temperature: 30 ℃.
Wavelength: 230nm.
Sample injection volume: 20. Mu.L.
Mobile phase: a linear gradient elution was performed according to Table 6 using 0.01% (v/v) phosphoric acid solution as mobile phase A and acetonitrile as mobile phase B.
TABLE 6
A diluent: acetonitrile
Peak order: praziquantel, clofenacin and milbexime A 3 Milbeoxime A 4 。
Milbeoxime control stock: 23.0mg of milbeoxime reference substance (source: USP, batch number: R086Q0, content: 97.4%) is precisely weighed, put into a 10mL measuring flask, dissolved and diluted to scale with a diluent, and shaken uniformly to obtain the final product.
Control solution (temporary fresh): 23mg of praziquantel reference substance (source: china medicine biological product testing institute, batch number: 100046-201205, content: 99.7%) and 46mg of clofen new reference substance (source: national pesticide quality supervision and inspection center (Shenyang), batch number: 20190601, content: 98.4%) are precisely weighed, 1.0mL of milbexime reference substance stock solution is precisely measured, placed in a 50mL measuring flask, dissolved and diluted to scale by a diluent, and shaken uniformly to obtain the finished product.
Test solution (prepared in the future): the concentration is consistent with that of the reference substance solution.
The invention is further illustrated by the following examples.
Example 1: system applicability
Chromatographic analysis procedure
And (5) performing a system applicability test according to the requirements of an analysis method. After the system was equilibrated, 20. Mu.L of the control solution was taken and injected into a liquid chromatograph, and the chromatogram was recorded. Relative Standard Deviation (RSD) was calculated by advancing 5-needle Std-1, and then by advancing 2-needle Std-2, and Std-2 was calculated with Std-1 for accuracy, as shown in Table 7.
TABLE 7
Equation for Std-1 to calculate Std-2 recovery:
A Std-1 mean of peak areas of each component in Std-1 chromatogram.
A Std-2 Mean of peak areas of each component in Std-2 color spectrum.
C Std-1 Concentration of each component in Std-1.
C Std-2 Concentration of each component in Std-2.
The results of the system suitability test are shown in tables 8 to 11.
TABLE 8
TABLE 9
Table 10
TABLE 11
It can be seen from the above that: the RSD of praziquantel 5 needle STD is 0.056%, which is far less than 2.0%, and the recovery rate of STD-2 to STD-1 is 99.4%; the RSD of the chlorfenacin 5-needle STD is 0.10 percent, which is far less than 2.0 percent, and the recovery rate of STD-2 to STD-1 is 100.3 percent; the RSD of the milbexime 5-needle STD is 0.075%, which is far less than 2.0%, and the recovery of STD-2 to STD-1 is 100.5%.
Example 2: specialization of
The specificity will be confirmed by the interference of the blank and the interference of the degraded impurities.
Blank solution (diluent): acetonitrile is taken for direct sample injection.
Sample: a similar weight of 14.2mg of a mixture of Praziquantel raw material (source: hebei Jiayi pharmaceutical Co., ltd., lot: JYC-200402), clofen raw material (source: zhejiang Cheng, lot: N181101), milbexime raw material (source: zhejiang Zhenzhi pharmaceutical Co., lot: 4037-A180405U) was weighed as a destructive test sample, respectively.
Acid degradation: weighing a sample, placing the sample in a 100mL measuring flask, adding 5.0mL of 1mol/L hydrochloric acid solution, standing at room temperature for 5min, adjusting the pH value to 7.0 by using 1mol/L sodium hydroxide solution, cooling to room temperature, diluting to a scale by using a diluent, and shaking uniformly to obtain the sample.
And (3) alkali degradation: weighing a sample, placing the sample in a 100mL measuring flask, adding 5.0mL of 1mol/L sodium hydroxide solution, standing at room temperature for 5min, adjusting the pH value to 7.0 by using 1mol/L hydrochloric acid solution, cooling to room temperature, diluting to a scale by using a diluent, and shaking uniformly to obtain the sample.
Oxidative degradation: weighing a sample, placing the sample into a 100mL measuring flask, adding 5.0mL of 30% hydrogen peroxide solution, immediately shaking at room temperature, diluting to a scale with a diluent, and shaking uniformly to obtain the product.
Thermal degradation: and (3) placing the sample in a transparent glass bottle for sealing, exposing the sample to a high temperature of 90 ℃ for 24 hours, cooling the sample to room temperature, and preparing a sample solution according to a sample solution preparation method.
High humidity degradation: the sample was sealed in a transparent glass bottle, exposed to 92.5% RH high humidity for 10 days, and then taken out, and a sample solution was prepared according to the sample solution preparation method.
The results of the proprietary study are shown in table 12.
Table 12
It can be seen from the above that: the blank solution does not appear a peak at the position of each main peak, and no interference exists; under all strong damage tests, the detected degradation peaks can be completely separated from all main peaks, and the separation degree of all main peaks and front and rear adjacent impurity peaks is more than 2.0; the milbemycin oxime acid degradation and alkali degradation conditions degrade by more than 30%, but the system applicability is good and no interference exists, which indicates that the method has good specificity.
Example 3: linearity of
The linear studies of praziquantel, clofenacin and milbexime in the content measurement gave concentrations ranging from 60 to 140%. The linear study was performed as follows.
Test program and result processing
The standard curve y=ax+b is plotted with the concentration (μg/mL) on the abscissa and the peak area on the ordinate.
And calculating a correlation coefficient r of the standard curve.
Calculating the linear deviation bias%: i.e. intercept b relative to peak area y at 100% concentration 100％ Is a percentage of (2).
Calculating an average response factor (MRF):
the results of the linearity test are shown in tables 13 to 15 and fig. 9 to 11.
TABLE 13
TABLE 14
TABLE 15
It can be seen from the above that: repetitive RSD of repeating three injections at each concentration n＝3 Less than 2.0%, indicating that the method has good system precision at all concentrations; correlation coefficient r: is 0.9997 (praziquantel), 0.9991 (clofenacin) and 1 (milbexime), which are all more than 0.999, which shows that the method has good linear regression; linear deviation Bias%: 1.7% (praziquantel), 1.8% (clofenacin) and 0.00041% (milbexime), all less than 2.0%, the systematic error of the method is smaller.
From the above, praziquantel has good linearity and system precision at the concentration of 283.20-654.64 mug/mL; the clofenacin has good linearity and system precision at the concentration of 570.44-1171.20 mug/mL; milbeoxime has good linearity and system precision at a concentration of 27.57-64.32 mug/mL.
Example 4: accuracy of
The accuracy of the method was evaluated by recovery after adding three components corresponding to 60%, 100% and 140% of the self-control concentration to the blank solution.
Milbeoxime control stock: accurately weighing 23.0mg of milbeoxime reference substance, placing into a 10mL measuring flask, dissolving with diluent, diluting to scale, and shaking.
Control solution: accurately weighing praziquantel reference substance 23mg and clofen new reference substance 46mg, accurately weighing 1.0mL of milbexime standard substance stock solution, placing into a 50mL measuring flask, dissolving with diluent, diluting to scale, and shaking.
Self-control stock: taking 230mg of praziquantel reference substance and 460mg of clofen new reference substance, precisely weighing, placing into a 50mL measuring flask, adding a diluent for dissolution, diluting to a scale, and shaking uniformly to obtain the praziquantel.
60% of the standard sample solution: precisely measuring 6.0mL of self control stock solution and 1.2mL of milbexime control stock solution, placing into a 100mL measuring flask, adding diluent to dilute to scale, and shaking uniformly. 3 parts in total.
100% labeled sample solution: precisely measuring 10.0mL of self-control stock solution and 2.0mL of milbexime control stock solution, placing into a 100mL measuring flask, adding diluent to dilute to scale, and shaking uniformly. 3 parts in total.
140% labeled sample solution: precisely measuring 14.0mL of self control stock solution and 2.8mL of milbexime control stock solution, placing into a 100mL measuring flask, adding diluent to dilute to scale, and shaking uniformly. 3 parts in total.
The standard recovery rates of the standard sample solutions of each stage are calculated according to the external standard method from tables 16 to 18 by repeating the sample injection of the reference solution 5 times and the rest solution 2 times according to the reference solution, 60% standard solution, 100% standard solution and 140% standard solution.
Table 16
TABLE 17
TABLE 18
It can be seen from the above that: standard recovery at 60%, 100%, 140% concentration: all are between 98.0 and 102.0 percent, which shows that the recovery rate is very good at all concentrations; RSD% of recovery from 3 trials at the same concentration n＝3 Are all much smaller than 2.0%; RSD% of average recovery between different concentrations n＝3 Are all much smaller than 2.0%.
From the above, it is clear that praziquantel at a concentration of 283.20-654.64. Mu.g/mL, clofenacin at a concentration of 570.44-1171.20. Mu.g/mL, milbexime at a concentration of 27.57-64.32. Mu.g/mL, namely, equivalent to 60-140% of the content, has good accuracy.
Example 5: precision of
Method reproducibility
6 parts of sample (mixture of three components of the same batch) solutions are prepared in parallel, and the three-component content detection is carried out according to the requirement of a detection method. In order to evaluate the repeatability of the results of the method on testing the same batch of samples under the same experimental environment.
Intermediate precision of the process
The 6 sample solutions used for the intermediate precision were prepared as required using the same lot number as the samples for the repeatability test. The test should be performed by different laboratory staff on different dates with different instruments, at intervals of at least 24 hours. In order to prove the reproducibility of the results of the method when testing the same batch of samples under a variable experimental environment. The results of the precision test are shown in Table 19.
TABLE 19
It can be seen from the above that: the same day method repeatability test: RSD% among the 6 detection results n＝6 No more than 0.42% and much less than 2.0%, indicating good reproducibility of the method. Repeat test by different testers on different days: of the total of 12 detection results, RSD% n＝12 The total content of the components is not more than 0.45 percent and is far less than 2.0 percent, which indicates that the method has good intermediate precision.
From the above, the method has good repeatability and precision.
Example 6: durability study
The key parameters are confirmed under the condition that one chromatographic parameter is changed and other factors (instruments and people) are unchanged, and if a large influence is found, the influence factors need to be strictly controlled in the analysis method setting.
Column temperature (. + -. 2 ℃ C.) was varied.
Wavelength is changed (+ -2 nm).
The flow rate was varied (+ -0.2 mL/min).
The column was changed (different manufacturers same specification: luna C18 (2) 250X 4.6mm,5 μm).
Chromatographic analysis procedure: each HPLC condition was tested for system suitability as described in the content analysis method, and then the content was determined for the same batch of samples. The system suitability durability test results are shown in Table 20.
Table 20
It can be seen from the above that: the difference between the measured content result under each change condition and the measured result under the initial condition is within 2.0%, the RSD of Std-1 peak area under each change condition is within 2.0%, and the recovery rate of Std-2 calculated by Std-1 under each change condition is between 98.0 and 102.0%. After the detection method is slightly changed, the consistency of the content detection result can be ensured, and the method for detecting the content of the three components has good durability.
Example 7: content method verification
The content detection method refers to European pharmacopoeia EP9.0 of new quality of veterinary clofen [4] And (5) making a standard. The method for verifying the content of the invention [5] 。
The main peak and each impurity peak were found to be well separated under the prescribed chromatographic conditions. The verification result shows that the method has good specificity, accuracy, linearity, durability and precision, and is suitable and accurate for detecting the content of the three components.
The summary of the results of the content verification is shown in Table 21.
Table 21 content methodology validation results table
As can be seen from Table 21, the results of the verification of the content analysis method show that the method has good specificity, accuracy, linearity, durability and precision, and is suitable and accurate for simultaneously detecting the content of three components.
Reference is made to:
[1] quality standard of praziquantel version EP9.0 of European pharmacopoeia.
[2] New quality standard of chlorofene version 43 of United states pharmacopoeia.
[3] Quality standard of milbezoxime praziquantel tablet in bulletin 2272 of Ministry of agriculture.
[4] New quality standard of veterinary clofen of European pharmacopoeia EP 9.0.
[5] The method for analyzing the quality standard of veterinary drugs in appendix 9101 of the edition of the Chinese veterinary pharmacopoeia 2020 verifies the guiding principle.
The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments. Those skilled in the art will appreciate that, in light of the principles of the present invention, improvements and modifications can be made without departing from the scope of the invention.
Claims (3)
1. A method for simultaneously detecting the contents of praziquantel, clofenacet and milbexime is characterized by comprising the following steps: the method comprises the following steps:
(1) Taking the diluent as a blank solution, injecting the blank solution into a liquid chromatograph, and recording chromatograms at wavelengths of 210nm, 230nm and 255 nm;
(2) Respectively weighing three components of praziquantel, clofenacin and milbexime, adding a diluent to prepare a solution containing 26 mug of praziquantel, 73 mug of clofenacin and 4.8 mug of milbexime as sample solutions per 1mL, injecting the sample solutions into a liquid chromatograph, and recording chromatograms at wavelengths of 210nm, 230nm and 255 nm;
(3) Taking a proper amount of each of praziquantel, praziquantel impurity A, praziquantel impurity B, praziquantel impurity C, clofenacin impurity B, clofenacin impurity C, clofenacin impurity G, milbexime and milbexime impurity D, adding a diluent to dissolve and prepare a mixed solution containing 1mg of each 1mL of each of the above-mentioned reference substances as a system applicability test solution, injecting the mixed solution into a liquid chromatograph, and recording chromatograms at wavelengths of 230nm and 255 nm;
in the step (3), the praziquantel impurity A is (11 bRS) -2-benzoyl-1, 2,3,6,7, 11B-hexahydro-4H-pyrazinyl [2,1-a ] isoquinolin-4-one, the praziquantel impurity B is 2- (cyclohexaformyl) -2,3,6, 7-tetrahydro-4H-pyrazino [2,1-a ] isoquinolin-4-one, and the praziquantel impurity C is N-formyl-N- [ 2-oxo-2- (1-oxo-3, 4-dihydroisoquinolin-2 (1H) -yl) ethyl ] cyclohexanecarboxamide;
in the step (3), the novel chlorofene impurity B is N- [ (2, 5-dichloro-4-p-hydroxybenzoyl) carbamoyl ] -2, 6-difluorobenzamide, the novel chlorofene impurity C is N- [ 3-chloro-4- (1, 2, 3-perfluorobutadienropoxy) phenyl-carbamoyl ] -2, 6-difluorobenzamide, and the novel chlorofene impurity G is 2, 5-dichloro-4- [3- (2, 6-difluorobenzoyl) ureido ] diphenyl carbonate;
in step (3), the milbeoxime impurity D is (1 ' R,2R,4' S,5S,6R,8' R,10' E,13' R,14' E,16' E,
20'r,21' z, 24's) -24' -hydroxy-21 '- (oximino) -6- (propane-2-yl) -5,11',13',22' tetramethyl-3, 4,5, 6-tetrahydrospiro [ pyran-2, 6'- [3,7,19] trioxytetracyclo [15.6.1.14,8.020,24] cyclopentadec [10,14,16,22] tetraen ] -2' -one;
in the step (1), the step (2) and the step (3), the chromatographic column in the liquid chromatograph is bonded silica gel by octadecylsilane;
mobile phase a is phosphoric acid solution and mobile phase B is acetonitrile.
2. The method for simultaneously detecting the three component contents of praziquantel, clofenacet and milbexime according to claim 1, wherein: in the step (1), the step (2) and the step (3), the diluent is acetonitrile.
3. The method for simultaneously detecting the three component contents of praziquantel, clofenacet and milbexime according to claim 1, wherein: in the step (1), the step (2) and the step (3), gradient elution is carried out in the liquid chromatograph, the flow rate is 1.0mL/min, the column temperature is 30 ℃, the ultraviolet detection wavelength is 230nm, and the sample injection amount is 20 mu L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111657868.5A CN114280200B (en) | 2021-12-30 | 2021-12-30 | Method for simultaneously detecting three component contents of praziquantel, clofenacin and milbexime |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111657868.5A CN114280200B (en) | 2021-12-30 | 2021-12-30 | Method for simultaneously detecting three component contents of praziquantel, clofenacin and milbexime |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114280200A CN114280200A (en) | 2022-04-05 |
| CN114280200B true CN114280200B (en) | 2023-07-07 |
Family
ID=80878900
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111657868.5A Active CN114280200B (en) | 2021-12-30 | 2021-12-30 | Method for simultaneously detecting three component contents of praziquantel, clofenacin and milbexime |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114280200B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2015202084A1 (en) * | 2010-04-02 | 2015-05-14 | Boehringer Ingelheim Animal Health USA Inc. | Parasiticidal compositions comprising multiple active agents, methods and uses thereof |
| CN106659160A (en) * | 2014-06-30 | 2017-05-10 | 日产化学工业株式会社 | Insecticide, miticide, nematicide, molluscicide, disinfectant, or bactericide composition, and pest control method |
| CN111208249A (en) * | 2020-01-14 | 2020-05-29 | 信阳农林学院 | Method for determining content of effective components of anthelmintic by high performance liquid chromatography |
-
2021
- 2021-12-30 CN CN202111657868.5A patent/CN114280200B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2015202084A1 (en) * | 2010-04-02 | 2015-05-14 | Boehringer Ingelheim Animal Health USA Inc. | Parasiticidal compositions comprising multiple active agents, methods and uses thereof |
| CN106659160A (en) * | 2014-06-30 | 2017-05-10 | 日产化学工业株式会社 | Insecticide, miticide, nematicide, molluscicide, disinfectant, or bactericide composition, and pest control method |
| CN111208249A (en) * | 2020-01-14 | 2020-05-29 | 信阳农林学院 | Method for determining content of effective components of anthelmintic by high performance liquid chromatography |
Non-Patent Citations (4)
| Title |
|---|
| An LC–MS Method for Determination of Milbemycin Oxime in Dog Plasma;Qianqian Xu 等;Journal of Chromatographic Science;999-1004 * |
| 猫经口内服2种米尔贝肟吡喹酮片的药代动力学比较研究;金之源 等;中国畜牧兽医;第41卷(第12期);157-160 * |
| 超高效液相色谱-串联质谱法同时检测牛乳中4种驱虫药物残留;侯改成 等;食品科学;第36卷(第06期);216-219 * |
| 超高效液相色谱测定柑橘中的虱螨脲、灭幼脲和氟啶脲;张月 等;现代农药;第13卷(第06期);41-43 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114280200A (en) | 2022-04-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Choi et al. | Glutamate neurotoxicity in cortical cell culture | |
| Ullian et al. | Invulnerability of retinal ganglion cells to NMDA excitotoxicity | |
| Staffen et al. | Measuring the progression of idiopathic Parkinson's disease with [123 I] β-CIT SPECT | |
| Anzalone et al. | Cortical cholinergic abnormalities contribute to the amnesic state induced by pyrithiamine‐induced thiamine deficiency in the rat | |
| Haughey et al. | Biosensor screening for veterinary drug residues in foodstuffs | |
| Ota et al. | Metformin prevents methylglyoxal-induced apoptosis of mouse Schwann cells | |
| Renner et al. | Cell-type-specific high throughput toxicity testing in human midbrain organoids | |
| Evans et al. | Development of an in vitro bioassay for measuring susceptibility to macrocyclic lactone anthelmintics in Dirofilaria immitis | |
| Raposo et al. | Evaluation of lacrimal production, osmolarity, crystallization, proteomic profile, and biochemistry of capuchin monkeys’ tear film | |
| CN116183771B (en) | Detection method of related substances in levofloxacin preparation | |
| Bavunoğlu et al. | Oleander poisoning as an example of self-medication attempt | |
| JP5303209B2 (en) | Evaluation method, screening method, and production method of substance that lowers blood glucose level | |
| CN114280200B (en) | Method for simultaneously detecting three component contents of praziquantel, clofenacin and milbexime | |
| Chen et al. | Simultaneous Determination of Malachite Green, Chloramphenicols, Sulfonamides, and Fluoroquinolones Residues in Fish by Liquid Chromatography‐Mass Spectrometry | |
| Zhu et al. | Acute and chronic toxicity of deltamethrin, permethrin, and dihaloacetylated heterocyclic pyrethroids in mice | |
| Jenkins et al. | A new primary screening test for anthelmintics utilizing the parasitic stages of Nippostrongylus brasiliensis, in vitro | |
| Du et al. | Azobenzene-avermectin B1a derivatives for optical modulation of insect behaviors | |
| Bennett et al. | Evaluation of transdermal application of glipizide in a pluronic lecithin gel to healthy cats | |
| Hacıoğlu et al. | Development and Validation of HPLC Method for determination of Nateglinide in Drug Substances | |
| CN105853386B (en) | Tablet containing dapagliflozin propylene glycol hydrate and preparation method thereof | |
| Jian Wang et al. | Systemic administration of domoic acid-induced spinal cord lesions in neonatal rats | |
| Vieira et al. | Dose–response and histopathological study, with special attention to the hypophysis, of the differential effects of domoic acid on rats and mice | |
| KR101189639B1 (en) | Oral formulations comprising metformin and &-glycosidase inhibitor and processes for the preparation thereof | |
| CN114527230A (en) | Method for detecting concentration of vitamin A and vitamin E in human dry blood spots | |
| US20090124018A1 (en) | Mix for quality control of the medicine "glycine tablets for sublingual applying o.1g." and its preparation method |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |