CN114031566A - Preparation method of stable isotope labeled diazepam internal standard reagent - Google Patents
Preparation method of stable isotope labeled diazepam internal standard reagent Download PDFInfo
- Publication number
- CN114031566A CN114031566A CN202110894605.XA CN202110894605A CN114031566A CN 114031566 A CN114031566 A CN 114031566A CN 202110894605 A CN202110894605 A CN 202110894605A CN 114031566 A CN114031566 A CN 114031566A
- Authority
- CN
- China
- Prior art keywords
- stable isotope
- diazepam
- isotope labeled
- reaction
- labeled
- 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.)
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- 229960003529 diazepam Drugs 0.000 title claims abstract description 46
- AAOVKJBEBIDNHE-UHFFFAOYSA-N diazepam Chemical compound N=1CC(=O)N(C)C2=CC=C(Cl)C=C2C=1C1=CC=CC=C1 AAOVKJBEBIDNHE-UHFFFAOYSA-N 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 23
- 238000006243 chemical reaction Methods 0.000 claims abstract description 42
- HXITXNWTGFUOAU-UHFFFAOYSA-N phenylboronic acid Chemical compound OB(O)C1=CC=CC=C1 HXITXNWTGFUOAU-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000000126 substance Substances 0.000 claims abstract description 19
- 239000002994 raw material Substances 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 13
- 239000000243 solution Substances 0.000 claims description 24
- 239000003960 organic solvent Substances 0.000 claims description 19
- 239000002904 solvent Substances 0.000 claims description 19
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 claims description 18
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 15
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 14
- 239000012074 organic phase Substances 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims description 12
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical class [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 238000004440 column chromatography Methods 0.000 claims description 12
- 238000005406 washing Methods 0.000 claims description 12
- ZUWXHHBROGLWNH-UHFFFAOYSA-N (2-amino-5-chlorophenyl)-phenylmethanone Chemical compound NC1=CC=C(Cl)C=C1C(=O)C1=CC=CC=C1 ZUWXHHBROGLWNH-UHFFFAOYSA-N 0.000 claims description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 10
- 238000001035 drying Methods 0.000 claims description 10
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
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- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 claims description 7
- JZWOKDTXYPEJEW-UHFFFAOYSA-N 7-chloro-5-phenyl-2,3-dihydro-1h-1,4-benzodiazepine Chemical compound C12=CC(Cl)=CC=C2NCCN=C1C1=CC=CC=C1 JZWOKDTXYPEJEW-UHFFFAOYSA-N 0.000 claims description 7
- PIBWKRNGBLPSSY-UHFFFAOYSA-L palladium(II) chloride Chemical compound Cl[Pd]Cl PIBWKRNGBLPSSY-UHFFFAOYSA-L 0.000 claims description 7
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- FKIIWCPNMDIJBG-UHFFFAOYSA-N 2-chloroacetamide (3-chlorophenyl)-phenylmethanone Chemical compound ClC=1C=CC=C(C(=O)C2=CC=CC=C2)C1.ClCC(=O)N FKIIWCPNMDIJBG-UHFFFAOYSA-N 0.000 claims description 6
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- NFHFRUOZVGFOOS-UHFFFAOYSA-N palladium;triphenylphosphane Chemical compound [Pd].C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1.C1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 NFHFRUOZVGFOOS-UHFFFAOYSA-N 0.000 claims description 6
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 6
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- 239000002253 acid Substances 0.000 claims description 4
- 150000003863 ammonium salts Chemical class 0.000 claims description 4
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 claims description 4
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- 239000000706 filtrate Substances 0.000 claims description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Substances [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 4
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- 238000010791 quenching Methods 0.000 claims description 4
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- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 claims description 4
- 239000008096 xylene Substances 0.000 claims description 4
- VGCXGMAHQTYDJK-UHFFFAOYSA-N Chloroacetyl chloride Chemical compound ClCC(Cl)=O VGCXGMAHQTYDJK-UHFFFAOYSA-N 0.000 claims description 3
- YZCKVEUIGOORGS-OUBTZVSYSA-N Deuterium Chemical compound [2H] YZCKVEUIGOORGS-OUBTZVSYSA-N 0.000 claims description 3
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-diisopropylethylamine Substances CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 3
- 235000019270 ammonium chloride Nutrition 0.000 claims description 3
- 229910052805 deuterium Inorganic materials 0.000 claims description 3
- VKYKSIONXSXAKP-UHFFFAOYSA-N hexamethylenetetramine Chemical compound C1N(C2)CN3CN1CN2C3 VKYKSIONXSXAKP-UHFFFAOYSA-N 0.000 claims description 3
- VUQUOGPMUUJORT-UHFFFAOYSA-N methyl 4-methylbenzenesulfonate Chemical compound COS(=O)(=O)C1=CC=C(C)C=C1 VUQUOGPMUUJORT-UHFFFAOYSA-N 0.000 claims description 3
- CYPYTURSJDMMMP-WVCUSYJESA-N (1e,4e)-1,5-diphenylpenta-1,4-dien-3-one;palladium Chemical compound [Pd].[Pd].C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1 CYPYTURSJDMMMP-WVCUSYJESA-N 0.000 claims description 2
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- JIVLDFFWTQYGSR-UHFFFAOYSA-N 4,7-dimethyl-[1,10]phenanthroline Chemical compound C1=CC2=C(C)C=CN=C2C2=C1C(C)=CC=N2 JIVLDFFWTQYGSR-UHFFFAOYSA-N 0.000 claims description 2
- DHDHJYNTEFLIHY-UHFFFAOYSA-N 4,7-diphenyl-1,10-phenanthroline Chemical compound C1=CC=CC=C1C1=CC=NC2=C1C=CC1=C(C=3C=CC=CC=3)C=CN=C21 DHDHJYNTEFLIHY-UHFFFAOYSA-N 0.000 claims description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 2
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 claims description 2
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- ROFVEXUMMXZLPA-UHFFFAOYSA-N Bipyridyl Chemical group N1=CC=CC=C1C1=CC=CC=N1 ROFVEXUMMXZLPA-UHFFFAOYSA-N 0.000 claims description 2
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 2
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 claims description 2
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 claims description 2
- RYXZOQOZERSHHQ-UHFFFAOYSA-N [2-(2-diphenylphosphanylphenoxy)phenyl]-diphenylphosphane Chemical compound C=1C=CC=C(P(C=2C=CC=CC=2)C=2C=CC=CC=2)C=1OC1=CC=CC=C1P(C=1C=CC=CC=1)C1=CC=CC=C1 RYXZOQOZERSHHQ-UHFFFAOYSA-N 0.000 claims description 2
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- 229940043376 ammonium acetate Drugs 0.000 claims description 2
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- LNAMMBFJMYMQTO-FNEBRGMMSA-N chloroform;(1e,4e)-1,5-diphenylpenta-1,4-dien-3-one;palladium Chemical class [Pd].[Pd].ClC(Cl)Cl.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1.C=1C=CC=CC=1\C=C\C(=O)\C=C\C1=CC=CC=C1 LNAMMBFJMYMQTO-FNEBRGMMSA-N 0.000 claims description 2
- KFGVRWGDTLZAAO-UHFFFAOYSA-N cyclopenta-1,3-diene dicyclohexyl(cyclopenta-1,3-dien-1-yl)phosphane iron(2+) Chemical compound [Fe++].c1cc[cH-]c1.C1CCC(CC1)P(C1CCCCC1)c1ccc[cH-]1 KFGVRWGDTLZAAO-UHFFFAOYSA-N 0.000 claims description 2
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- 208000032140 Sleepiness Diseases 0.000 description 1
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- 238000009825 accumulation Methods 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000001773 anti-convulsant effect Effects 0.000 description 1
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- 239000002249 anxiolytic agent Substances 0.000 description 1
- 230000000949 anxiolytic effect Effects 0.000 description 1
- 229940125717 barbiturate Drugs 0.000 description 1
- HNYOPLTXPVRDBG-UHFFFAOYSA-N barbituric acid Chemical compound O=C1CC(=O)NC(=O)N1 HNYOPLTXPVRDBG-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- ULELPEYHAFTGRE-UHFFFAOYSA-N bromobenzene formic acid Chemical class C(=O)O.BrC1=CC=CC=C1 ULELPEYHAFTGRE-UHFFFAOYSA-N 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
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- 230000007547 defect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- VAYGXNSJCAHWJZ-UHFFFAOYSA-N dimethyl sulfate Chemical compound COS(=O)(=O)OC VAYGXNSJCAHWJZ-UHFFFAOYSA-N 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000009313 farming Methods 0.000 description 1
- 238000005755 formation reaction Methods 0.000 description 1
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 235000010299 hexamethylene tetramine Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- -1 hypnotic Substances 0.000 description 1
- 239000003326 hypnotic agent Substances 0.000 description 1
- 238000010813 internal standard method Methods 0.000 description 1
- INQOMBQAUSQDDS-UHFFFAOYSA-N iodomethane Chemical compound IC INQOMBQAUSQDDS-UHFFFAOYSA-N 0.000 description 1
- 210000003715 limbic system Anatomy 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000003158 myorelaxant agent Substances 0.000 description 1
- KJONHKAYOJNZEC-UHFFFAOYSA-N nitrazepam Chemical compound C12=CC([N+](=O)[O-])=CC=C2NC(=O)CN=C1C1=CC=CC=C1 KJONHKAYOJNZEC-UHFFFAOYSA-N 0.000 description 1
- 229960001454 nitrazepam Drugs 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 150000002923 oximes Chemical class 0.000 description 1
- 229960000762 perphenazine Drugs 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 229960002244 promethazine hydrochloride Drugs 0.000 description 1
- XXPDBLUZJRXNNZ-UHFFFAOYSA-N promethazine hydrochloride Chemical compound Cl.C1=CC=C2N(CC(C)N(C)C)C3=CC=CC=C3SC2=C1 XXPDBLUZJRXNNZ-UHFFFAOYSA-N 0.000 description 1
- 239000012217 radiopharmaceutical Substances 0.000 description 1
- 229940121896 radiopharmaceutical Drugs 0.000 description 1
- 230000002799 radiopharmaceutical effect Effects 0.000 description 1
- 238000006722 reduction reaction Methods 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000002040 relaxant effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000001624 sedative effect Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- 231100001274 therapeutic index Toxicity 0.000 description 1
- 229960004098 thioridazine hydrochloride Drugs 0.000 description 1
- NZFNXWQNBYZDAQ-UHFFFAOYSA-N thioridazine hydrochloride Chemical compound Cl.C12=CC(SC)=CC=C2SC2=CC=CC=C2N1CCC1CCCCN1C NZFNXWQNBYZDAQ-UHFFFAOYSA-N 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
- C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
- C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
- C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
- C07D243/16—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals
- C07D243/18—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals substituted in position 2 by nitrogen, oxygen or sulfur atoms
- C07D243/24—Oxygen atoms
- C07D243/26—Preparation from compounds already containing the benzodiazepine skeleton
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
- C07B59/002—Heterocyclic compounds
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D243/00—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms
- C07D243/06—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4
- C07D243/10—Heterocyclic compounds containing seven-membered rings having two nitrogen atoms as the only ring hetero atoms having the nitrogen atoms in positions 1 and 4 condensed with carbocyclic rings or ring systems
- C07D243/14—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines
- C07D243/16—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals
- C07D243/18—1,4-Benzodiazepines; Hydrogenated 1,4-benzodiazepines substituted in position 5 by aryl radicals substituted in position 2 by nitrogen, oxygen or sulfur atoms
- C07D243/24—Oxygen atoms
- C07D243/28—Preparation including building-up the benzodiazepine skeleton from compounds containing no hetero rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/05—Isotopically modified compounds, e.g. labelled
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- Chemical & Material Sciences (AREA)
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention relates to a preparation method of a stable isotope labeled diazepam internal standard reagent for food safety detection, belonging to the field of research and development of standard substances for food safety detection. The stable isotope labeled diazepam is obtained by using stable isotope labeled phenylboronic acid as a raw material through catalytic addition, acylation, cyclization, methylation, and finally separation and purification. The process has the advantages of simple and easily obtained raw materials required by synthesis, mild reaction conditions, avoidance of use of a highly toxic methylation reagent, capability of ensuring that the chemical purity of the target product stable isotope labeled diazepam can reach more than 98 percent and the isotope abundance can reach more than 98 percent, and capability of meeting the technical requirements of a stable isotope labeled internal standard reagent required by a liquid chromatography tandem mass spectrometry for food safety detection.
Description
Technical Field
The invention relates to a preparation method of a stable isotope labeled diazepam internal standard reagent for food safety detection, belonging to the field of research and development of standard substances for food safety detection.
Background
Diazepam is commonly known as diazepam and is benzodiazepine
The compounds have anxiolytic, sedative, hypnotic, anticonvulsant, antiepileptic and central muscle relaxant effects. The medicine is prepared fromThe compound hypnotic can selectively act on the limbic system of the brain, is combined with a central BDZ receptor to promote the release of gamma-aminobutyric acid or a synaptic transmission function, can induce sleep in larger dose, has the characteristics of high therapeutic index, small influence on respiration, little influence on fast wave sleep, no influence on liver drug enzymes, no anesthesia in large dose and the like compared with barbiturate hypnotics, and is the most common hypnotic in clinic at present. Although the half-life period of the drug is short, metabolites still have activity, so the drug has toxic action on human bodies after excessive residual accumulation. In 2017, 10 and 27, the worldwide health organization international cancer research institution publishes a carcinogen list for preliminary reference arrangement, and diazepam is in a category 3 carcinogen list. Some breeding and feed production enterprises pursue high profit for one side, illegally add the medicine in the production link, especially in livestock breeding, the feeding of diazepam can cause nerve paralysis and quiet somnolence so as to reduce the activity of livestock and achieve the aims of rapidly fattening and shortening the marketing time. However, the livestock and poultry fed with diazepam have the medicine residue, and the poisoning can be caused after the human beings take the medicine and accumulate the medicine for a long time. It has been totally banned from detection in foods of animal origin and use of diazepam in farming has been banned in several countries and regions.
At present, the No. 176 bulletin of Ministry of agriculture has stipulated that the use of benzodiazepines in animal feed and drinking water is strictly prohibited
A quasi drug. The detection method of the related medicine residue comprises the following steps: the detection standard in the feed is the high performance liquid chromatography and liquid chromatography mass spectrometry combined method for synchronously measuring promethazine hydrochloride, chlorpromazine hydrochloride, diazepam, thioridazine hydrochloride and perphenazine in NY/T1458-2007 feed published by Ministry of agriculture and the benzodiazepine in NY/T1757-2009 feed
Determination of quasi-drugs by liquid chromatography-tandem mass spectrometry; the related detection standard method in the food is the residual quantity of the sedative drug in the SN/T2113-2008 import and export animal derived food published by the customs administrationThe detection method comprises a liquid chromatogram-mass spectrum/mass spectrometry method and a liquid chromatogram-mass spectrum/mass spectrometry method for determining the residue of the benzodiazepine drug in the SN/T3847-2019 export food. Wherein the determination of the residual quantity of benzodiazepine drugs in SN/T3847 and 2019 export foods by a liquid chromatography-mass spectrometry/mass spectrometry method adopts a stable isotope internal standard method for quantification and uses diazepam (phenyl-D)5) As an isotopic internal standard.
At present, diazepam labeled by stable isotopes is only provided by foreign standard product companies and can only be solved by import, thereby severely restricting the application of the compound in domestic food safety detection. Up to now, the literature on the Synthesis of stable isotope-labeled Diazepam is Allison F.Fentiman, which is published in 1976, Synthesis of bacterium-labelled drugs of use for use as internal standards in the quantification by selection of microorganisms II Deutrium-labelled benzodiazepines, Chlordizaeposition, Demethyidizepam, Diazepam, and Oxazepam [ J.J.]Journal of laboratory Compounds and Radiopharmaceuticals, 1977, 13(4): 579-. This document is obtained by starting from 2-amino-5-chlorobenzoic acid and carrying out 8-step reactions such as acetylation, Grignard reaction, hydrolysis, oxime formation, cyclization, hydrolysis, methylation, reduction and the like. The related domestic invention patents include Duhongyan and Xuxiaoing published in 2017 (CN 107365276B), a preparation method of diazepam-D5 (CN 107522667B), and diazepam-D8And a method for preparing the same. The method comprises the steps of taking 6-chloro-2-methyl-4H-3, 1-benzoxazine-4-ketone as a raw material, reacting with a deuterated bromobenzene format reagent, hydrolyzing, acylating, cyclizing and methylating to obtain the stable diazepam-D5And diazepam-D8. The methods have the defects of long synthesis steps, strict requirements on the conditions for preparing stable isotope labeling format reagents, high technical difficulty, use of highly toxic and carcinogenic methylating reagents such as dimethyl sulfate or methyl iodide and the like.
Therefore, the development of a preparation method of stable isotope labeled diazepam, which has the advantages of short reaction route, simple operation, mild conditions, avoidance of use of a highly toxic methylation reagent, high total yield and low cost, is urgently needed to meet the increasing application requirement of the compound in the aspect of food safety detection in China.
Disclosure of Invention
The invention aims to provide a preparation method of stable isotope labeled diazepam suitable for food safety detection. The synthesis method has the advantages of easily obtained raw materials, short reaction route, simple operation, mild conditions and no use of toxic and carcinogenic methylation reagents; the final product has high total yield, high chemical purity and undiluted isotopic abundance, and effectively reduces the product cost.
The purpose of the invention can be realized by the following technical scheme: 1. a preparation method of a stable isotope Labeled Diazepam internal standard reagent is disclosed, wherein the chemical structure of the stable isotope Labeled Diazepam internal standard reagent is shown as the formula (Labeled-Diazepam):
wherein R is H or D12C or13C; the preparation method is characterized by comprising the following general preparation steps:
wherein R is H or D12C or13C, the preparation steps are as follows:
(1) under the existence of a catalyst, a ligand, an additive and an organic solvent, adding stable isotope labeled phenylboronic acid, then adding 2-amino-5-chlorobenzonitrile, and reacting for 12-48 hours at the temperature of 30-120 ℃; after TLC tracking reaction raw materials completely react, quenching the reaction mixed solution with water, filtering, extracting the filtrate with an organic solvent, combining organic phases, washing with a saturated sodium bicarbonate solution and a saturated sodium chloride solution in sequence, adding a drying agent for drying, removing the solvent by reduced pressure distillation, and carrying out column chromatography on residues to obtain the stable isotope labeled 2-amino-5-chlorobenzophenone;
the reaction of this step is as follows:
wherein the chemical structure of the stable isotope labeled phenylboronic acid is shown as formula (I), and the chemical structure of the stable isotope labeled 2-amino-5-chlorobenzophenone is shown as formula (II), wherein R is H or D12C or13C；
(2) Dissolving stable isotope labeled 2-amino-5-chlorobenzophenone in an organic solvent, adding chloroacetyl chloride, and reacting at 20-80 ℃ for 1-10 hours; after TLC tracking reaction raw materials completely react, quenching the reaction mixed solution with water, filtering, extracting the filtrate with an organic solvent, combining organic phases, washing with a saturated sodium bicarbonate solution and a saturated sodium chloride solution in sequence, adding a drying agent for drying, removing the solvent by reduced pressure distillation, and carrying out column chromatography on residues to obtain the stable isotope labeled 2-chloroacetamide-5-chlorobenzophenone;
the reaction of this step is as follows:
wherein the chemical structure of the stable isotope labeled 2-chloroacetamide-5-chlorobenzophenone is shown as the formula (III), wherein R is H or D12C or13C；
(3) Dissolving stable isotope labeled 2-chloroacetamide-5-chlorobenzophenone in an organic solvent, adding urotropine and ammonium salt, and reacting at 30-120 ℃ for 1-8 hours; after TLC tracks that the reaction raw materials react completely, the reaction mixed solution is dried by spinning, water is added, organic solvent is used for extraction, organic phases are combined, and the organic phases are washed by saturated sodium bicarbonate solution and saturated sodium chloride solution in turn, and then drying agent is added; distilling under reduced pressure to remove solvent, and subjecting the residue to column chromatography to obtain stable isotope labeled 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
-a 2-ketone;
the reaction of this step is as follows:
wherein the stable isotope is labeled with 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
The chemical structure of the-2-ketone is shown as a formula (IV), wherein R is H or D12C or13C；
(4) Labeling stable isotope with 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
Dissolving-2-ketone in an organic solvent, adding methyl p-toluenesulfonate and an acid-binding agent, and reacting for 2-12 hours at the temperature of 0-100 ℃; TLC tracks the reaction raw materials to react completely, then water is added after filtration, organic solvent is used for extraction, organic phases are combined, saturated sodium bicarbonate solution is used for washing, saturated sodium chloride solution is used for washing, and drying agent is added for drying; and (4) distilling under reduced pressure to remove the solvent, and carrying out column chromatography on the residue to obtain a pure product to obtain the stable isotope labeled diazepam.
The reaction of this step is as follows:
wherein the chemical structure of the stable isotope Labeled Diazepam is shown as the formula (Labeled-Diazepam), wherein R is H or D12C or13C。
In the step (1), the catalyst is one or more of palladium acetate, palladium chloride, bis (triphenylphosphine) palladium dichloride, tetrakis (triphenylphosphine) palladium, bis (diphenylphosphinophenyl ether) palladium dichloride, bis (diphenylphosphinoferrocene) palladium dichloride, palladium acetylacetonate, palladium trifluoroacetate, bis (diphenylphosphinofethanpalladium dichloride), tris (dibenzylideneacetone) dipalladium and tris (dibenzylideneacetone) dipalladium chloroform adductSeed growing; the ligand is one or more of 2,2 ' -bipyridyl, 4 ' -dimethyl-2, 2 ' -bipyridyl, 5 ' -dimethyl-2, 2 ' -bipyridyl, 4 ' -dimethoxy-2, 2 ' -bipyridyl, 1, 10-phenanthroline, 4, 7-dimethyl-1, 10-phenanthroline and 4, 7-diphenyl-1, 10-phenanthroline; the additive is one or more of hydrochloric acid, sulfuric acid, glacial acetic acid, formic acid, trifluoroacetic acid, dichloroacetic acid, methane sulfonic acid, p-toluenesulfonic acid and benzoic acid; the solvent is one or more of tetrahydrofuran, dioxane, toluene, xylene, 2-methyltetrahydrofuran, N-dimethylformamide, methanol, ethanol, isopropanol, N-propanol, benzene and chlorobenzene; the stable isotope labeled phenylboronic acid is deuterium labeled phenylboronic acid,13C-labelled phenylboronic acid or deuterium and13c-ditag phenylboronic acid; the reaction temperature is 30-120 ℃; the reaction time is 12-48 hours.
In the step (2), the organic solvent is one or more of tetrahydrofuran, 2-methyltetrahydrofuran, dioxane, benzene, toluene, xylene, acetonitrile and N, N-dimethylformamide; the reaction temperature is 20-80 ℃; the reaction time is 1-10 hours.
In the step (3), the organic solvent is one or more of methanol, ethanol, n-propanol and isopropanol; the ammonium salt is one or more of ammonium chloride, ammonium formate, ammonium acetate and ammonium sulfate; the reaction temperature is 30-120 ℃; the reaction time is 1-8 hours.
In the step (4), the organic solvent is one or more of chloroform, dichloromethane, N-dimethylformamide, toluene, tetrahydrofuran and dioxane; the acid-binding agent is one or more of triethylamine, N-diisopropylethylamine, 4-dimethylaminopyridine, triethanolamine, sodium bicarbonate, sodium carbonate, potassium bicarbonate, potassium carbonate and cesium carbonate; the reaction temperature is 0-100 ℃; the reaction time is 2-12 hours.
Compared with the prior art, the invention has the beneficial effects that: the synthetic method has the advantages of simple and easily-obtained raw materials, short synthetic steps, simple operation, mild reaction conditions, avoidance of use of highly toxic and carcinogenic methylation reagents, and effective reduction of personnel health and environmental pollution risks. The stable isotope labeled nitrazepam for food safety detection obtained by the preparation method has important significance for relevant food safety supervision and detection work in China.
The preparation method of the stable isotope labeled oxazepam internal standard reagent for monitoring the clinical treatment medicine has the advantages of simple and easily obtained raw materials, mild reaction conditions, high chemical purity of the final product and high total yield, effectively reduces the risks of personnel health and environmental pollution, and basically does not dilute the isotope abundance. The preparation route and the preparation method of the stable isotope labeled diazepam are not reported in documents, the chemical purity of the prepared stable isotope labeled diazepam is more than 98%, and the isotope abundance is more than 98%. The stable isotope labeled diazepam obtained by the preparation method meets the requirements of an internal standard reagent for liquid chromatography tandem mass spectrometry detection, and has important significance for food safety supervision and detection work related in China.
Description of the drawings:
FIG. 1: 2-amino-5-chlorobenzophenone-D5Is/are as follows1H NMR chart
FIG. 2: 2-amino-5-chlorobenzophenone-D5LC-MS diagram of
FIG. 3: 2-chloroacetamido-5-chlorobenzophenone (phenyl-D)5) Is/are as follows1H NMR chart
FIG. 4: 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
-2-one (phenyl-D)5) Is/are as follows1H NMR chart
FIG. 5: 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
-2-one (phenyl-D)5) LC-MS diagram of
FIG. 6: diazepam (phenyl-D)5) Is/are as follows1H NMR chart
FIG. 7: diazepam (phenyl-D)5) Is/are as follows13C NMR chart
FIG. 8: diazepam (phenyl-D)5) HRMS diagram of
FIG. 9: diazepam (phenyl)-D5) UPLC diagram of
Detailed Description
The present invention will be further described with reference to the following examples and the accompanying drawings, which are not intended to limit the scope of the present invention in any way.
Example 1: 2-amino-5-chlorobenzophenone (phenyl-D)5) Preparation of
In a 500mL three-necked flask, deuterated phenylboronic acid (12.7g, 0.1mol), 2-amino-5-chlorobenzophenone (7.63g, 0.05mol), the catalyst tetrakis (triphenylphosphine) palladium (2.89g, 2.5mmol), the ligand 4,4 '-dimethyl-2, 2' -bipyridine (920mg, 5mmol), the additive trifluoroacetic acid (57g, 0.5mol), and tetrahydrofuran were added: water 1: 1 (250 mL); reacting at 90 ℃ for 24 hours; TLC (thin layer chromatography) is used for tracking reaction raw materials to completely react, the reaction liquid is cooled to room temperature, 200mL of water is added, ethyl acetate (3X 300mL) is used for extraction, organic phases are combined, the mixture is washed by saturated sodium bicarbonate solution and saturated sodium chloride solution in sequence, anhydrous sodium sulfate is added for drying, the solvent is removed through reduced pressure distillation, and the residue is subjected to column chromatography to obtain the stable isotope labeled 2-amino-5-chlorobenzophenone (phenyl-D)5) (10.9g, yield 92%).1H NMR(DMSO-D 6600 MHz): δ 7.34(dd, 1H), 7.21(s, 2H), 7.19(d, 1H), 6.90(d, 1H), see fig. 1; LC-MS (ESI +)237[ M + H]+See fig. 2.
Example 2: 2-chloroacetamido-5-chlorobenzophenone (phenyl-D)5) Preparation of
In a 100mL three-necked flask, 2-amino-5-chlorobenzophenone (phenyl-D) was added5) (2.37g, 10mmol), chloroacetyl chloride (1.69g, 15mmol) and dried dioxane (20 mL); introducing nitrogen, and reacting for 12 hours at the temperature of 20 ℃; after TLC tracking reaction raw material completely reacts, reduced pressure distillation is carried outRemoving solvent, adding 20mL of water, extracting with ethyl acetate (3 × 30mL), mixing organic phases, sequentially washing with saturated sodium bicarbonate solution, washing with saturated sodium chloride solution, drying with anhydrous sodium sulfate, distilling under reduced pressure to remove solvent, and subjecting the residue to column chromatography to obtain stable isotope labeled 2-chloroacetamido-5-chlorobenzophenone (phenyl-D)5) (2.97g, yield 95%).1H NMR(DMSO-D 6600 MHz): δ 10.50(s, 1H), 7.70(d, 2H), 7.44(t, 1H), 4.06(s, 1H), see fig. 3.
Example 3: 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
-2-one (phenyl-D)5) Preparation of
Adding stable isotope labeled 2-chloroacetamido-5-chlorobenzophenone (phenyl-D) into a 50mL three-necked bottle5) (3.13g, 10mmol), urotropin (2.80g, 20mmol), ammonium chloride (1.07g, 20mmol), methanol (20 mL); heating to reflux under the protection of nitrogen, and reacting at the temperature for 24 hours; TLC (thin layer chromatography) is used for tracking the complete reaction of the reaction raw materials, the solvent is removed by distillation under reduced pressure, 20mL of water is added, ethyl acetate (3X 30mL) is used for extraction, organic phases are combined, the mixture is washed by saturated sodium bicarbonate solution and saturated sodium chloride solution in turn, anhydrous sodium sulfate is added for drying, the solvent is removed by distillation under reduced pressure, and the residue is subjected to column chromatography to obtain the stable isotope labeled 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
-2-one (phenyl-D)5) (2.97g, yield 95%).1H NMR(DMSO-D6600 MHz): δ 9.41(s, 1H), 7.45(dd, 1H), 7.30(dd, 1H), 7.15(dd, 1H), see FIG. 4; LC-MS (ESI +)276[ M + H]+See fig. 5.
Example 4: diazepam (phenyl-D)5) Preparation of
Adding 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine labeled by stable isotope into a 100mL three-necked bottle
-2-one (phenyl-D)5) (1.38g, 5mmol), methyl p-toluenesulfonate (1.12g, 6mmol), N-diisopropylethylamine (1.29g, 10mmol), dichloromethane (20 mL); reacting for 8 hours at 40 ℃ under the protection of nitrogen; TLC (thin layer chromatography) tracking reaction raw materials, adding 30mL of water after complete reaction, extracting with dichloromethane (3X 30mL), combining organic phases, washing with saturated sodium bicarbonate solution, washing with saturated sodium chloride solution, adding anhydrous sodium sulfate for drying, distilling under reduced pressure to remove solvent, and carrying out column chromatography on residues to obtain the stable isotope labeled diazepam (phenyl-D)5) (1.32g, yield 91%).1HNMR(DMSO-D6600 MHz): δ 7.75(dd, J ═ 24Hz, 1H), 7.64(d, J ═ 18Hz, 1H), 7.23(d, J ═ 6Hz, 1H), 4.60(d, J ═ 18Hz, 1H), 3.81(d, J ═ 18Hz, 1H), 3.31(s, 1H), see fig. 6 for details;13C NMR(DMSO-D6150 MHz): δ 169.68, 168.51, 143.05, 138.23, 131.91, 129.94, 129.34, 128.22, 124.35, 57.15, 34.77, as detailed in fig. 7; HRMS (ESI +)290.10947[ M + H ]]See fig. 8 for details; the chemical purity by HPLC was 99.36%, as detailed in FIG. 9. Isotopic abundance 98.70% D, detailed in table 1.
TABLE 1 diazepam-D5Isotopic abundance of
The quality standard meets the requirement of a stable isotope internal standard reagent required by a liquid chromatogram tandem mass spectrometry for food safety detection.
13C-labeled Stable isotope-labeled diazepam Synthesis procedure as in the previous examplesLabeling diazepam with an agent (phenyl-D)5) The synthetic route of (1).
The above description is only for the purpose of illustrating the preferred embodiments of the present invention, and the present invention should not be limited by the disclosure of the preferred embodiments. Therefore, it is intended that all equivalents and modifications which do not depart from the spirit of the invention disclosed herein are deemed to be within the scope of the invention.
Claims (5)
1. A preparation method of a stable isotope Labeled Diazepam internal standard reagent is disclosed, wherein the chemical structure of the stable isotope Labeled Diazepam internal standard reagent is shown as the formula (Labeled-Diazepam):
wherein R is H or D12C or13C; the preparation method is characterized by comprising the following general preparation steps:
wherein R is H or D12C or13C, the preparation steps are as follows:
(1) under the existence of a catalyst, a ligand, an additive and an organic solvent, adding stable isotope labeled phenylboronic acid, then adding 2-amino-5-chlorobenzonitrile, and reacting for 12-48 hours at the temperature of 30-120 ℃; after TLC tracking reaction raw materials completely react, quenching the reaction mixed solution with water, filtering, extracting the filtrate with an organic solvent, combining organic phases, washing with a saturated sodium bicarbonate solution and a saturated sodium chloride solution in sequence, adding a drying agent for drying, removing the solvent by reduced pressure distillation, and carrying out column chromatography on residues to obtain the stable isotope labeled 2-amino-5-chlorobenzophenone;
the reaction of this step is as follows:
wherein the chemical structure of the stable isotope labeled phenylboronic acid is shown as formula (I), and the chemical structure of the stable isotope labeled 2-amino-5-chlorobenzophenone is shown as formula (II), wherein R is H or D12C or13C；
(2) Dissolving stable isotope labeled 2-amino-5-chlorobenzophenone in an organic solvent, adding chloroacetyl chloride, and reacting at 20-80 ℃ for 1-10 hours; after TLC tracking reaction raw materials completely react, quenching the reaction mixed solution with water, filtering, extracting the filtrate with an organic solvent, combining organic phases, washing with a saturated sodium bicarbonate solution and a saturated sodium chloride solution in sequence, adding a drying agent for drying, removing the solvent by reduced pressure distillation, and carrying out column chromatography on residues to obtain the stable isotope labeled 2-chloroacetamide-5-chlorobenzophenone;
the reaction of this step is as follows:
wherein the chemical structure of the stable isotope labeled 2-chloroacetamide-5-chlorobenzophenone is shown as the formula (III), wherein R is H or D12C or13C；
(3) Dissolving stable isotope labeled 2-chloroacetamide-5-chlorobenzophenone in an organic solvent, adding urotropine and ammonium salt, and reacting at 30-120 ℃ for 1-8 hours; after TLC tracks that the reaction raw materials react completely, the reaction mixed solution is dried by spinning, water is added, organic solvent is used for extraction, organic phases are combined, and the organic phases are washed by saturated sodium bicarbonate solution and saturated sodium chloride solution in turn, and then drying agent is added; distilling under reduced pressure to remove solvent, and subjecting the residue to column chromatography to obtain stable isotope labeled 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
-a 2-ketone;
the reaction of this step is as follows:
wherein the stable isotope is labeled with 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
The chemical structure of the-2-ketone is shown as a formula (IV), wherein R is H or D12C or13C；
(4) Labeling stable isotope with 7-chloro-1, 3-dihydro-5-phenyl-2H-1, 4-benzodiazepine
Dissolving-2-ketone in an organic solvent, adding methyl p-toluenesulfonate and an acid-binding agent, and reacting for 2-12 hours at the temperature of 0-100 ℃; TLC tracks the reaction raw materials to react completely, then water is added after filtration, organic solvent is used for extraction, organic phases are combined, saturated sodium bicarbonate solution is used for washing, saturated sodium chloride solution is used for washing, and drying agent is added for drying; and (4) distilling under reduced pressure to remove the solvent, and carrying out column chromatography on the residue to obtain a pure product to obtain the stable isotope labeled diazepam.
The reaction of this step is as follows:
wherein the chemical structure of the stable isotope Labeled Diazepam is shown as the formula (Labeled-Diazepam), wherein R is H or D12C or13C。
2. The method for preparing a stable isotope labeling diazepam internal standard reagent according to claim 1, wherein in the step (1), the catalyst is palladium acetate, palladium chloride, bis (triphenylphosphine) palladium dichloride, tetrakis (triphenylphosphine) palladium, bis (diphenylphosphinophenyl ether) palladium dichloride, bis (diphenylphosphinoferrocene) palladium dichloride, palladium acetylacetonate, palladium trifluoroacetate, bis (diphenyl phosphine ferrocene) palladium dichlorideOne or more of phenylphosphine) ethane palladium dichloride, tris (dibenzylideneacetone) dipalladium and tris (dibenzylideneacetone) dipalladium chloroform adducts; the ligand is one or more of 2,2 ' -bipyridyl, 4 ' -dimethyl-2, 2 ' -bipyridyl, 5 ' -dimethyl-2, 2 ' -bipyridyl, 4 ' -dimethoxy-2, 2 ' -bipyridyl, 1, 10-phenanthroline, 4, 7-dimethyl-1, 10-phenanthroline and 4, 7-diphenyl-1, 10-phenanthroline; the additive is one or more of hydrochloric acid, sulfuric acid, glacial acetic acid, formic acid, trifluoroacetic acid, dichloroacetic acid, methane sulfonic acid, p-toluenesulfonic acid and benzoic acid; the solvent is one or more of tetrahydrofuran, dioxane, toluene, xylene, 2-methyltetrahydrofuran, N-dimethylformamide, methanol, ethanol, isopropanol, N-propanol, benzene and chlorobenzene; the stable isotope labeled phenylboronic acid is deuterium labeled phenylboronic acid,13One of the C-labeled phenylboronic acids; the reaction temperature is 30-120 ℃; the reaction time is 12-48 hours. .
3. The method for preparing a stable isotope labeling diazepam internal standard reagent according to claim 1, wherein in the step (2), the solvent is one or more of tetrahydrofuran, 2-methyltetrahydrofuran, dioxane, benzene, toluene, xylene, acetonitrile and N, N-dimethylformamide; the reaction temperature is 20-80 ℃; the reaction time is 1-10 hours. .
4. The method for preparing a stable isotope labeling diazepam internal standard reagent according to claim 1, wherein in the step (3), the solvent is one or more of methanol, ethanol, n-propanol and isopropanol; the ammonium salt is one or more of ammonium chloride, ammonium formate, ammonium acetate and ammonium sulfate; the reaction temperature is 30-120 ℃; the reaction time is 1-8 hours.
5. The method for preparing a stable isotope labeling diazepam internal standard reagent according to claim 1, wherein in the step (4), the solvent is one or more of chloroform, dichloromethane, N-dimethylformamide, toluene, tetrahydrofuran and dioxane; the acid-binding agent is one or more of triethylamine, N-diisopropylethylamine, 4-dimethylaminopyridine, triethanolamine, sodium bicarbonate, sodium carbonate, potassium bicarbonate, potassium carbonate and cesium carbonate; the reaction temperature is 0-100 ℃; the reaction time is 2-12 hours.
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| CN107501196A (en) * | 2017-08-08 | 2017-12-22 | 公安部物证鉴定中心 | Intermediate for preparing diazepam D5 and diazepam D8 and preparation method thereof |
| CN107522667A (en) * | 2017-08-08 | 2017-12-29 | 公安部物证鉴定中心 | Diazepam D8 and preparation method thereof |
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