CN110456087A - 一种舍曲林检测试剂及其制备和使用方法 - Google Patents
一种舍曲林检测试剂及其制备和使用方法 Download PDFInfo
- Publication number
- CN110456087A CN110456087A CN201910767565.5A CN201910767565A CN110456087A CN 110456087 A CN110456087 A CN 110456087A CN 201910767565 A CN201910767565 A CN 201910767565A CN 110456087 A CN110456087 A CN 110456087A
- Authority
- CN
- China
- Prior art keywords
- sertraline
- solution
- added
- reagent
- mass fraction
- 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.)
- Granted
Links
- 229960002073 sertraline Drugs 0.000 title claims abstract description 186
- VGKDLMBJGBXTGI-SJCJKPOMSA-N sertraline Chemical compound C1([C@@H]2CC[C@@H](C3=CC=CC=C32)NC)=CC=C(Cl)C(Cl)=C1 VGKDLMBJGBXTGI-SJCJKPOMSA-N 0.000 title claims abstract description 127
- 239000003153 chemical reaction reagent Substances 0.000 title claims abstract description 72
- 238000001514 detection method Methods 0.000 title claims abstract description 44
- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- 102000004190 Enzymes Human genes 0.000 claims abstract description 62
- 108090000790 Enzymes Proteins 0.000 claims abstract description 62
- -1 Sertraline maleimide derivatives Chemical class 0.000 claims abstract description 37
- 230000005847 immunogenicity Effects 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 78
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 36
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 36
- 150000001875 compounds Chemical class 0.000 claims description 36
- 239000000203 mixture Substances 0.000 claims description 30
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 22
- 239000002904 solvent Substances 0.000 claims description 21
- 238000006243 chemical reaction Methods 0.000 claims description 20
- 238000003018 immunoassay Methods 0.000 claims description 19
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 18
- 238000000746 purification Methods 0.000 claims description 18
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 17
- 230000002163 immunogen Effects 0.000 claims description 17
- 102100031126 6-phosphogluconolactonase Human genes 0.000 claims description 16
- 108010029731 6-phosphogluconolactonase Proteins 0.000 claims description 16
- 108010018962 Glucosephosphate Dehydrogenase Proteins 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 16
- 230000015572 biosynthetic process Effects 0.000 claims description 15
- 239000000872 buffer Substances 0.000 claims description 15
- 238000003786 synthesis reaction Methods 0.000 claims description 15
- 239000012043 crude product Substances 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- 239000002253 acid Substances 0.000 claims description 13
- 239000000758 substrate Substances 0.000 claims description 13
- 238000010171 animal model Methods 0.000 claims description 12
- 239000005457 ice water Substances 0.000 claims description 12
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 12
- 108010077055 methylated bovine serum albumin Proteins 0.000 claims description 12
- 230000011987 methylation Effects 0.000 claims description 12
- 238000007069 methylation reaction Methods 0.000 claims description 12
- 239000011541 reaction mixture Substances 0.000 claims description 10
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 9
- 239000007983 Tris buffer Substances 0.000 claims description 9
- 238000002347 injection Methods 0.000 claims description 9
- 239000007924 injection Substances 0.000 claims description 9
- 239000005416 organic matter Substances 0.000 claims description 9
- 239000000741 silica gel Substances 0.000 claims description 9
- 229910002027 silica gel Inorganic materials 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 9
- 241000283707 Capra Species 0.000 claims description 8
- 229910052681 coesite Inorganic materials 0.000 claims description 8
- 229910052906 cristobalite Inorganic materials 0.000 claims description 8
- 239000000377 silicon dioxide Substances 0.000 claims description 8
- 229910052682 stishovite Inorganic materials 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 8
- 229910052905 tridymite Inorganic materials 0.000 claims description 8
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 238000000605 extraction Methods 0.000 claims description 7
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 7
- 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 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 6
- XPDWGBQVDMORPB-UHFFFAOYSA-N Fluoroform Chemical compound FC(F)F XPDWGBQVDMORPB-UHFFFAOYSA-N 0.000 claims description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 6
- WMFOQBRAJBCJND-UHFFFAOYSA-M Lithium hydroxide Chemical compound [Li+].[OH-] WMFOQBRAJBCJND-UHFFFAOYSA-M 0.000 claims description 6
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 claims description 6
- 239000007832 Na2SO4 Substances 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 6
- 239000002671 adjuvant Substances 0.000 claims description 6
- 150000001412 amines Chemical class 0.000 claims description 6
- 239000007864 aqueous solution Substances 0.000 claims description 6
- 239000012267 brine Substances 0.000 claims description 6
- 150000001718 carbodiimides Chemical class 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 235000019253 formic acid Nutrition 0.000 claims description 6
- 229950006238 nadide Drugs 0.000 claims description 6
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims description 6
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 claims description 6
- 230000035479 physiological effects, processes and functions Effects 0.000 claims description 6
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 6
- FGIUAXJPYTZDNR-UHFFFAOYSA-N potassium nitrate Chemical compound [K+].[O-][N+]([O-])=O FGIUAXJPYTZDNR-UHFFFAOYSA-N 0.000 claims description 6
- 239000008213 purified water Substances 0.000 claims description 6
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N sodium azide Substances [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims description 6
- 229910052938 sodium sulfate Inorganic materials 0.000 claims description 6
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000003643 water by type Substances 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- 102000014914 Carrier Proteins Human genes 0.000 claims description 4
- 108010078791 Carrier Proteins Proteins 0.000 claims description 4
- 101710088194 Dehydrogenase Proteins 0.000 claims description 4
- 229910019142 PO4 Inorganic materials 0.000 claims description 4
- 210000004369 blood Anatomy 0.000 claims description 4
- 239000008280 blood Substances 0.000 claims description 4
- 230000006837 decompression Effects 0.000 claims description 4
- 238000004090 dissolution Methods 0.000 claims description 4
- 239000010452 phosphate Substances 0.000 claims description 4
- 210000002381 plasma Anatomy 0.000 claims description 4
- 239000012460 protein solution Substances 0.000 claims description 4
- 210000002966 serum Anatomy 0.000 claims description 4
- CCMKPCBRNXKTKV-UHFFFAOYSA-N 1-hydroxy-5-sulfanylidenepyrrolidin-2-one Chemical compound ON1C(=O)CCC1=S CCMKPCBRNXKTKV-UHFFFAOYSA-N 0.000 claims description 3
- YOETUEMZNOLGDB-UHFFFAOYSA-N 2-methylpropyl carbonochloridate Chemical compound CC(C)COC(Cl)=O YOETUEMZNOLGDB-UHFFFAOYSA-N 0.000 claims description 3
- WDBQJSCPCGTAFG-QHCPKHFHSA-N 4,4-difluoro-N-[(1S)-3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-pyridin-3-ylpropyl]cyclohexane-1-carboxamide Chemical compound FC1(CCC(CC1)C(=O)N[C@@H](CCN1CCC(CC1)N1C(=NN=C1C)C(C)C)C=1C=NC=CC=1)F WDBQJSCPCGTAFG-QHCPKHFHSA-N 0.000 claims description 3
- BWGRDBSNKQABCB-UHFFFAOYSA-N 4,4-difluoro-N-[3-[3-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)-8-azabicyclo[3.2.1]octan-8-yl]-1-thiophen-2-ylpropyl]cyclohexane-1-carboxamide Chemical compound CC(C)C1=NN=C(C)N1C1CC2CCC(C1)N2CCC(NC(=O)C1CCC(F)(F)CC1)C1=CC=CS1 BWGRDBSNKQABCB-UHFFFAOYSA-N 0.000 claims description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 3
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- 239000007821 HATU Substances 0.000 claims description 3
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical compound O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 claims description 3
- LFZAGIJXANFPFN-UHFFFAOYSA-N N-[3-[4-(3-methyl-5-propan-2-yl-1,2,4-triazol-4-yl)piperidin-1-yl]-1-thiophen-2-ylpropyl]acetamide Chemical compound C(C)(C)C1=NN=C(N1C1CCN(CC1)CCC(C=1SC=CC=1)NC(C)=O)C LFZAGIJXANFPFN-UHFFFAOYSA-N 0.000 claims description 3
- 238000010266 Sephadex chromatography Methods 0.000 claims description 3
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical class OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 3
- 150000004075 acetic anhydrides Chemical class 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 229910021529 ammonia Inorganic materials 0.000 claims description 3
- 239000000908 ammonium hydroxide Substances 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- 238000000502 dialysis Methods 0.000 claims description 3
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 229910052742 iron Inorganic materials 0.000 claims description 3
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L magnesium chloride Substances [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 claims description 3
- 229910001629 magnesium chloride Inorganic materials 0.000 claims description 3
- QAWFLJGZSZIZHO-UHFFFAOYSA-N methyl 4-bromobutanoate Chemical class COC(=O)CCCBr QAWFLJGZSZIZHO-UHFFFAOYSA-N 0.000 claims description 3
- 230000003647 oxidation Effects 0.000 claims description 3
- 238000007254 oxidation reaction Methods 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N phosphoric acid Substances OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 3
- 239000011574 phosphorus Substances 0.000 claims description 3
- 229910052698 phosphorus Inorganic materials 0.000 claims description 3
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 3
- 235000010333 potassium nitrate Nutrition 0.000 claims description 3
- 239000004323 potassium nitrate Substances 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 239000012264 purified product Substances 0.000 claims description 3
- 239000000376 reactant Substances 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 239000011780 sodium chloride Substances 0.000 claims description 3
- 235000011121 sodium hydroxide Nutrition 0.000 claims description 3
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical class [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 3
- 235000010288 sodium nitrite Nutrition 0.000 claims description 3
- 238000005303 weighing Methods 0.000 claims description 3
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims description 2
- 102000002322 Egg Proteins Human genes 0.000 claims description 2
- 108010000912 Egg Proteins Proteins 0.000 claims description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 2
- 241001494479 Pecora Species 0.000 claims description 2
- 102000009843 Thyroglobulin Human genes 0.000 claims description 2
- 108010034949 Thyroglobulin Proteins 0.000 claims description 2
- 238000002835 absorbance Methods 0.000 claims description 2
- 230000008859 change Effects 0.000 claims description 2
- 229940126214 compound 3 Drugs 0.000 claims description 2
- 229940125898 compound 5 Drugs 0.000 claims description 2
- 235000014103 egg white Nutrition 0.000 claims description 2
- 210000000969 egg white Anatomy 0.000 claims description 2
- 108060003552 hemocyanin Proteins 0.000 claims description 2
- 150000003949 imides Chemical class 0.000 claims description 2
- 238000011534 incubation Methods 0.000 claims description 2
- 229920001184 polypeptide Polymers 0.000 claims description 2
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 2
- 102000004169 proteins and genes Human genes 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 210000003296 saliva Anatomy 0.000 claims description 2
- 229960002175 thyroglobulin Drugs 0.000 claims description 2
- 210000002700 urine Anatomy 0.000 claims description 2
- 238000004458 analytical method Methods 0.000 claims 1
- 238000003818 flash chromatography Methods 0.000 claims 1
- 238000002156 mixing Methods 0.000 claims 1
- 230000009870 specific binding Effects 0.000 claims 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid group Chemical class S(O)(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 10
- 230000035945 sensitivity Effects 0.000 abstract description 5
- 238000002474 experimental method Methods 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 238000003556 assay Methods 0.000 abstract description 2
- 239000012472 biological sample Substances 0.000 abstract description 2
- 230000003053 immunization Effects 0.000 abstract description 2
- 238000002649 immunization Methods 0.000 abstract description 2
- 239000000523 sample Substances 0.000 description 8
- 239000005556 hormone Substances 0.000 description 7
- 229940088597 hormone Drugs 0.000 description 7
- 238000005259 measurement Methods 0.000 description 7
- 229940079593 drug Drugs 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 6
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 229940076279 serotonin Drugs 0.000 description 3
- KWGRBVOPPLSCSI-WPRPVWTQSA-N (-)-ephedrine Chemical compound CN[C@@H](C)[C@H](O)C1=CC=CC=C1 KWGRBVOPPLSCSI-WPRPVWTQSA-N 0.000 description 2
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 description 2
- KWTSXDURSIMDCE-QMMMGPOBSA-N (S)-amphetamine Chemical compound C[C@H](N)CC1=CC=CC=C1 KWTSXDURSIMDCE-QMMMGPOBSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- DNXHEGUUPJUMQT-CBZIJGRNSA-N Estrone Chemical compound OC1=CC=C2[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 DNXHEGUUPJUMQT-CBZIJGRNSA-N 0.000 description 2
- QGXBDMJGAMFCBF-UHFFFAOYSA-N Etiocholanolone Natural products C1C(O)CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CCC21 QGXBDMJGAMFCBF-UHFFFAOYSA-N 0.000 description 2
- HEMJJKBWTPKOJG-UHFFFAOYSA-N Gemfibrozil Chemical compound CC1=CC=C(C)C(OCCCC(C)(C)C(O)=O)=C1 HEMJJKBWTPKOJG-UHFFFAOYSA-N 0.000 description 2
- BHHGXPLMPWCGHP-UHFFFAOYSA-N Phenethylamine Chemical compound NCCC1=CC=CC=C1 BHHGXPLMPWCGHP-UHFFFAOYSA-N 0.000 description 2
- 235000008331 Pinus X rigitaeda Nutrition 0.000 description 2
- 235000011613 Pinus brutia Nutrition 0.000 description 2
- 241000018646 Pinus brutia Species 0.000 description 2
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 2
- UCTWMZQNUQWSLP-UHFFFAOYSA-N adrenaline Chemical compound CNCC(O)C1=CC=C(O)C(O)=C1 UCTWMZQNUQWSLP-UHFFFAOYSA-N 0.000 description 2
- 150000001408 amides Chemical class 0.000 description 2
- VIROVYVQCGLCII-UHFFFAOYSA-N amobarbital Chemical compound CC(C)CCC1(CC)C(=O)NC(=O)NC1=O VIROVYVQCGLCII-UHFFFAOYSA-N 0.000 description 2
- 229940025084 amphetamine Drugs 0.000 description 2
- AEMFNILZOJDQLW-QAGGRKNESA-N androst-4-ene-3,17-dione Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CCC2=C1 AEMFNILZOJDQLW-QAGGRKNESA-N 0.000 description 2
- 229960005471 androstenedione Drugs 0.000 description 2
- AEMFNILZOJDQLW-UHFFFAOYSA-N androstenedione Natural products O=C1CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 AEMFNILZOJDQLW-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 238000010219 correlation analysis Methods 0.000 description 2
- 229960003638 dopamine Drugs 0.000 description 2
- 229960003627 gemfibrozil Drugs 0.000 description 2
- QRMZSPFSDQBLIX-UHFFFAOYSA-N homovanillic acid Chemical compound COC1=CC(CC(O)=O)=CC=C1O QRMZSPFSDQBLIX-UHFFFAOYSA-N 0.000 description 2
- JYGXADMDTFJGBT-VWUMJDOOSA-N hydrocortisone Chemical compound O=C1CC[C@]2(C)[C@H]3[C@@H](O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 JYGXADMDTFJGBT-VWUMJDOOSA-N 0.000 description 2
- 150000002576 ketones Chemical class 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229960002748 norepinephrine Drugs 0.000 description 2
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 description 2
- DHHVAGZRUROJKS-UHFFFAOYSA-N phentermine Chemical compound CC(C)(N)CC1=CC=CC=C1 DHHVAGZRUROJKS-UHFFFAOYSA-N 0.000 description 2
- 239000013062 quality control Sample Substances 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- KDYFGRWQOYBRFD-UHFFFAOYSA-N succinic acid Chemical compound OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- DNXHEGUUPJUMQT-UHFFFAOYSA-N (+)-estrone Natural products OC1=CC=C2C3CCC(C)(C(CC4)=O)C4C3CCC2=C1 DNXHEGUUPJUMQT-UHFFFAOYSA-N 0.000 description 1
- UIKROCXWUNQSPJ-VIFPVBQESA-N (-)-cotinine Chemical compound C1CC(=O)N(C)[C@@H]1C1=CC=CN=C1 UIKROCXWUNQSPJ-VIFPVBQESA-N 0.000 description 1
- SCPADBBISMMJAW-UHFFFAOYSA-N (10S)-3c.17t-Dihydroxy-10r.13c-dimethyl-17c-((R)-1-hydroxy-aethyl)-(5tH.8cH.9tH.14tH)-hexadecahydro-1H-cyclopenta[a]phenanthren Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(O)C)(O)C1(C)CC2 SCPADBBISMMJAW-UHFFFAOYSA-N 0.000 description 1
- PROQIPRRNZUXQM-UHFFFAOYSA-N (16alpha,17betaOH)-Estra-1,3,5(10)-triene-3,16,17-triol Natural products OC1=CC=C2C3CCC(C)(C(C(O)C4)O)C4C3CCC2=C1 PROQIPRRNZUXQM-UHFFFAOYSA-N 0.000 description 1
- RDJGLLICXDHJDY-NSHDSACASA-N (2s)-2-(3-phenoxyphenyl)propanoic acid Chemical compound OC(=O)[C@@H](C)C1=CC=CC(OC=2C=CC=CC=2)=C1 RDJGLLICXDHJDY-NSHDSACASA-N 0.000 description 1
- METKIMKYRPQLGS-GFCCVEGCSA-N (R)-atenolol Chemical compound CC(C)NC[C@@H](O)COC1=CC=C(CC(N)=O)C=C1 METKIMKYRPQLGS-GFCCVEGCSA-N 0.000 description 1
- MURXBMUMXOJVCR-UHFFFAOYSA-N 1,2-bis($l^{1}-oxidanyl)ethane Chemical compound [O]CC[O] MURXBMUMXOJVCR-UHFFFAOYSA-N 0.000 description 1
- ICBPURKUPVLVCM-UHFFFAOYSA-N 1,5-dimethyl-2-phenylpyrazol-3-one;2-hydroxy-2-phenylacetic acid Chemical compound OC(=O)C(O)C1=CC=CC=C1.CN1C(C)=CC(=O)N1C1=CC=CC=C1 ICBPURKUPVLVCM-UHFFFAOYSA-N 0.000 description 1
- FUFLCEKSBBHCMO-UHFFFAOYSA-N 11-dehydrocorticosterone Natural products O=C1CCC2(C)C3C(=O)CC(C)(C(CC4)C(=O)CO)C4C3CCC2=C1 FUFLCEKSBBHCMO-UHFFFAOYSA-N 0.000 description 1
- ZESRJSPZRDMNHY-YFWFAHHUSA-N 11-deoxycorticosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 ZESRJSPZRDMNHY-YFWFAHHUSA-N 0.000 description 1
- NVKAWKQGWWIWPM-ABEVXSGRSA-N 17-β-hydroxy-5-α-Androstan-3-one Chemical compound C1C(=O)CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CC[C@H]21 NVKAWKQGWWIWPM-ABEVXSGRSA-N 0.000 description 1
- DBPWSSGDRRHUNT-CEGNMAFCSA-N 17α-hydroxyprogesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@@](C(=O)C)(O)[C@@]1(C)CC2 DBPWSSGDRRHUNT-CEGNMAFCSA-N 0.000 description 1
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 1
- 150000003923 2,5-pyrrolediones Chemical class 0.000 description 1
- NGBBVGZWCFBOGO-UHFFFAOYSA-N 3,4-Methylenedioxyamphetamine Chemical compound CC(N)CC1=CC=C2OCOC2=C1 NGBBVGZWCFBOGO-UHFFFAOYSA-N 0.000 description 1
- GMLREHXYJDLZOU-LEPYJNQMSA-N 3-Acetylmorphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4OC(C)=O GMLREHXYJDLZOU-LEPYJNQMSA-N 0.000 description 1
- LKQDFQLSEHWIRK-UKBVDAKRSA-N 3alpha,17alpha-Dihydroxy-5beta-pregnan-20-one Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@@](C(=O)C)(O)[C@@]2(C)CC1 LKQDFQLSEHWIRK-UKBVDAKRSA-N 0.000 description 1
- QGXBDMJGAMFCBF-BNSUEQOYSA-N 3alpha-hydroxy-5beta-androstan-17-one Chemical compound C1[C@H](O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC[C@@H]21 QGXBDMJGAMFCBF-BNSUEQOYSA-N 0.000 description 1
- BMUKKTUHUDJSNZ-UHFFFAOYSA-N 4-[1-hydroxy-2-(1-phenoxypropan-2-ylamino)propyl]phenol Chemical compound C=1C=C(O)C=CC=1C(O)C(C)NC(C)COC1=CC=CC=C1 BMUKKTUHUDJSNZ-UHFFFAOYSA-N 0.000 description 1
- SCPADBBISMMJAW-UHHUKTEYSA-N 5beta-Pregnane-3alpha,17alpha,20alpha-triol Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@@]([C@@H](O)C)(O)[C@@]2(C)CC1 SCPADBBISMMJAW-UHHUKTEYSA-N 0.000 description 1
- QGXBDMJGAMFCBF-HLUDHZFRSA-N 5α-Androsterone Chemical compound C1[C@H](O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC[C@H]21 QGXBDMJGAMFCBF-HLUDHZFRSA-N 0.000 description 1
- 108060003345 Adrenergic Receptor Proteins 0.000 description 1
- 102000017910 Adrenergic receptor Human genes 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- PQSUYGKTWSAVDQ-ZVIOFETBSA-N Aldosterone Chemical compound C([C@@]1([C@@H](C(=O)CO)CC[C@H]1[C@@H]1CC2)C=O)[C@H](O)[C@@H]1[C@]1(C)C2=CC(=O)CC1 PQSUYGKTWSAVDQ-ZVIOFETBSA-N 0.000 description 1
- PQSUYGKTWSAVDQ-UHFFFAOYSA-N Aldosterone Natural products C1CC2C3CCC(C(=O)CO)C3(C=O)CC(O)C2C2(C)C1=CC(=O)CC2 PQSUYGKTWSAVDQ-UHFFFAOYSA-N 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- KLYCPFXDDDMZNQ-UHFFFAOYSA-N Benzyne Chemical compound C1=CC#CC=C1 KLYCPFXDDDMZNQ-UHFFFAOYSA-N 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- VBGLYOIFKLUMQG-UHFFFAOYSA-N Cannabinol Chemical compound C1=C(C)C=C2C3=C(O)C=C(CCCCC)C=C3OC(C)(C)C2=C1 VBGLYOIFKLUMQG-UHFFFAOYSA-N 0.000 description 1
- MFYSYFVPBJMHGN-ZPOLXVRWSA-N Cortisone Chemical compound O=C1CC[C@]2(C)[C@H]3C(=O)C[C@](C)([C@@](CC4)(O)C(=O)CO)[C@@H]4[C@@H]3CCC2=C1 MFYSYFVPBJMHGN-ZPOLXVRWSA-N 0.000 description 1
- MFYSYFVPBJMHGN-UHFFFAOYSA-N Cortisone Natural products O=C1CCC2(C)C3C(=O)CC(C)(C(CC4)(O)C(=O)CO)C4C3CCC2=C1 MFYSYFVPBJMHGN-UHFFFAOYSA-N 0.000 description 1
- UIKROCXWUNQSPJ-UHFFFAOYSA-N Cotinine Natural products C1CC(=O)N(C)C1C1=CC=CN=C1 UIKROCXWUNQSPJ-UHFFFAOYSA-N 0.000 description 1
- MQJKPEGWNLWLTK-UHFFFAOYSA-N Dapsone Chemical compound C1=CC(N)=CC=C1S(=O)(=O)C1=CC=C(N)C=C1 MQJKPEGWNLWLTK-UHFFFAOYSA-N 0.000 description 1
- PHMBVCPLDPDESM-YWIQKCBGSA-N Ecgonine Natural products C1[C@H](O)[C@@H](C(O)=O)[C@H]2CC[C@@H]1N2C PHMBVCPLDPDESM-YWIQKCBGSA-N 0.000 description 1
- 102000005915 GABA Receptors Human genes 0.000 description 1
- 108010005551 GABA Receptors Proteins 0.000 description 1
- 102000004300 GABA-A Receptors Human genes 0.000 description 1
- 108090000839 GABA-A Receptors Proteins 0.000 description 1
- JMBQKKAJIKAWKF-UHFFFAOYSA-N Glutethimide Chemical compound C=1C=CC=CC=1C1(CC)CCC(=O)NC1=O JMBQKKAJIKAWKF-UHFFFAOYSA-N 0.000 description 1
- 240000008669 Hedera helix Species 0.000 description 1
- 102000000543 Histamine Receptors Human genes 0.000 description 1
- 108010002059 Histamine Receptors Proteins 0.000 description 1
- HEFNNWSXXWATRW-UHFFFAOYSA-N Ibuprofen Chemical compound CC(C)CC1=CC=C(C(C)C(O)=O)C=C1 HEFNNWSXXWATRW-UHFFFAOYSA-N 0.000 description 1
- PWWVAXIEGOYWEE-UHFFFAOYSA-N Isophenergan Chemical compound C1=CC=C2N(CC(C)N(C)C)C3=CC=CC=C3SC2=C1 PWWVAXIEGOYWEE-UHFFFAOYSA-N 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- XADCESSVHJOZHK-UHFFFAOYSA-N Meperidine Chemical compound C=1C=CC=CC=1C1(C(=O)OCC)CCN(C)CC1 XADCESSVHJOZHK-UHFFFAOYSA-N 0.000 description 1
- 102000010909 Monoamine Oxidase Human genes 0.000 description 1
- 108010062431 Monoamine oxidase Proteins 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- GIYXAJPCNFJEHY-UHFFFAOYSA-N N-methyl-3-phenyl-3-[4-(trifluoromethyl)phenoxy]-1-propanamine hydrochloride (1:1) Chemical compound Cl.C=1C=CC=CC=1C(CCNC)OC1=CC=C(C(F)(F)F)C=C1 GIYXAJPCNFJEHY-UHFFFAOYSA-N 0.000 description 1
- CMWTZPSULFXXJA-UHFFFAOYSA-N Naproxen Natural products C1=C(C(C)C(O)=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-UHFFFAOYSA-N 0.000 description 1
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 1
- ONBWJWYUHXVEJS-ZTYRTETDSA-N Normorphine Chemical compound C([C@@H](NCC1)[C@@H]2C=C[C@@H]3O)C4=CC=C(O)C5=C4[C@@]21[C@H]3O5 ONBWJWYUHXVEJS-ZTYRTETDSA-N 0.000 description 1
- BRUQQQPBMZOVGD-XFKAJCMBSA-N Oxycodone Chemical compound O=C([C@@H]1O2)CC[C@@]3(O)[C@H]4CC5=CC=C(OC)C2=C5[C@@]13CCN4C BRUQQQPBMZOVGD-XFKAJCMBSA-N 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- CZWCKYRVOZZJNM-UHFFFAOYSA-N Prasterone sodium sulfate Natural products C1C(OS(O)(=O)=O)CCC2(C)C3CCC(C)(C(CC4)=O)C4C3CC=C21 CZWCKYRVOZZJNM-UHFFFAOYSA-N 0.000 description 1
- YWYQTGBBEZQBGO-BERLURQNSA-N Pregnanediol Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](O)C)[C@@]2(C)CC1 YWYQTGBBEZQBGO-BERLURQNSA-N 0.000 description 1
- ORNBQBCIOKFOEO-YQUGOWONSA-N Pregnenolone Natural products O=C(C)[C@@H]1[C@@]2(C)[C@H]([C@H]3[C@@H]([C@]4(C)C(=CC3)C[C@@H](O)CC4)CC2)CC1 ORNBQBCIOKFOEO-YQUGOWONSA-N 0.000 description 1
- 206010039897 Sedation Diseases 0.000 description 1
- 206010041349 Somnolence Diseases 0.000 description 1
- CYQFCXCEBYINGO-UHFFFAOYSA-N THC Natural products C1=C(C)CCC2C(C)(C)OC3=CC(CCCCC)=CC(O)=C3C21 CYQFCXCEBYINGO-UHFFFAOYSA-N 0.000 description 1
- 208000001871 Tachycardia Diseases 0.000 description 1
- 206010044565 Tremor Diseases 0.000 description 1
- YWYQTGBBEZQBGO-UHFFFAOYSA-N UC1011 Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(O)C)C1(C)CC2 YWYQTGBBEZQBGO-UHFFFAOYSA-N 0.000 description 1
- 229960001138 acetylsalicylic acid Drugs 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- NDAUXUAQIAJITI-UHFFFAOYSA-N albuterol Chemical compound CC(C)(C)NCC(O)C1=CC=C(O)C(CO)=C1 NDAUXUAQIAJITI-UHFFFAOYSA-N 0.000 description 1
- 229960002478 aldosterone Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 229940098184 amytal Drugs 0.000 description 1
- 229960003473 androstanolone Drugs 0.000 description 1
- 229940061641 androsterone Drugs 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 229960002274 atenolol Drugs 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- YXKTVDFXDRQTKV-HNNXBMFYSA-N benzphetamine Chemical compound C([C@H](C)N(C)CC=1C=CC=CC=1)C1=CC=CC=C1 YXKTVDFXDRQTKV-HNNXBMFYSA-N 0.000 description 1
- 229960002837 benzphetamine Drugs 0.000 description 1
- ZTGXAWYVTLUPDT-UHFFFAOYSA-N cannabidiol Natural products OC1=CC(CCCCC)=CC(O)=C1C1C(C(C)=C)CC=C(C)C1 ZTGXAWYVTLUPDT-UHFFFAOYSA-N 0.000 description 1
- 229960003453 cannabinol Drugs 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- ZPEIMTDSQAKGNT-UHFFFAOYSA-N chlorpromazine Chemical compound C1=C(Cl)C=C2N(CCCN(C)C)C3=CC=CC=C3SC2=C1 ZPEIMTDSQAKGNT-UHFFFAOYSA-N 0.000 description 1
- 229960001076 chlorpromazine Drugs 0.000 description 1
- 230000001713 cholinergic effect Effects 0.000 description 1
- 230000003153 cholinolytic effect Effects 0.000 description 1
- 125000000490 cinnamyl group Chemical group C(C=CC1=CC=CC=C1)* 0.000 description 1
- 229960004362 clorazepate Drugs 0.000 description 1
- XDDJGVMJFWAHJX-UHFFFAOYSA-M clorazepic acid anion Chemical compound C12=CC(Cl)=CC=C2NC(=O)C(C(=O)[O-])N=C1C1=CC=CC=C1 XDDJGVMJFWAHJX-UHFFFAOYSA-M 0.000 description 1
- 229940125782 compound 2 Drugs 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000004246 corpus luteum Anatomy 0.000 description 1
- 229960004544 cortisone Drugs 0.000 description 1
- 229950006073 cotinine Drugs 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- PHMBVCPLDPDESM-UHFFFAOYSA-N d-Pseudoekgonin Natural products C1C(O)C(C(O)=O)C2CCC1N2C PHMBVCPLDPDESM-UHFFFAOYSA-N 0.000 description 1
- KWGRBVOPPLSCSI-UHFFFAOYSA-N d-ephedrine Natural products CNC(C)C(O)C1=CC=CC=C1 KWGRBVOPPLSCSI-UHFFFAOYSA-N 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- CZWCKYRVOZZJNM-USOAJAOKSA-N dehydroepiandrosterone sulfate Chemical compound C1[C@@H](OS(O)(=O)=O)CC[C@]2(C)[C@H]3CC[C@](C)(C(CC4)=O)[C@@H]4[C@@H]3CC=C21 CZWCKYRVOZZJNM-USOAJAOKSA-N 0.000 description 1
- CYQFCXCEBYINGO-IAGOWNOFSA-N delta1-THC Chemical compound C1=C(C)CC[C@H]2C(C)(C)OC3=CC(CCCCC)=CC(O)=C3[C@@H]21 CYQFCXCEBYINGO-IAGOWNOFSA-N 0.000 description 1
- 229960003654 desoxycortone Drugs 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- UZBQIPPOMKBLAS-UHFFFAOYSA-N diethylazanide Chemical compound CC[N-]CC UZBQIPPOMKBLAS-UHFFFAOYSA-N 0.000 description 1
- XYYVYLMBEZUESM-UHFFFAOYSA-N dihydrocodeine Natural products C1C(N(CCC234)C)C2C=CC(=O)C3OC2=C4C1=CC=C2OC XYYVYLMBEZUESM-UHFFFAOYSA-N 0.000 description 1
- ZZVUWRFHKOJYTH-UHFFFAOYSA-N diphenhydramine Chemical compound C=1C=CC=CC=1C(OCCN(C)C)C1=CC=CC=C1 ZZVUWRFHKOJYTH-UHFFFAOYSA-N 0.000 description 1
- 229960000520 diphenhydramine Drugs 0.000 description 1
- 208000002173 dizziness Diseases 0.000 description 1
- DLNKOYKMWOXYQA-UHFFFAOYSA-N dl-pseudophenylpropanolamine Natural products CC(N)C(O)C1=CC=CC=C1 DLNKOYKMWOXYQA-UHFFFAOYSA-N 0.000 description 1
- 230000003291 dopaminomimetic effect Effects 0.000 description 1
- 229960004242 dronabinol Drugs 0.000 description 1
- PHMBVCPLDPDESM-FKSUSPILSA-N ecgonine Chemical compound C1[C@H](O)[C@H](C(O)=O)[C@H]2CC[C@@H]1N2C PHMBVCPLDPDESM-FKSUSPILSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229960002179 ephedrine Drugs 0.000 description 1
- 229960005309 estradiol Drugs 0.000 description 1
- 229930182833 estradiol Natural products 0.000 description 1
- 229960001348 estriol Drugs 0.000 description 1
- PROQIPRRNZUXQM-ZXXIGWHRSA-N estriol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H]([C@H](O)C4)O)[C@@H]4[C@@H]3CCC2=C1 PROQIPRRNZUXQM-ZXXIGWHRSA-N 0.000 description 1
- 229960003399 estrone Drugs 0.000 description 1
- 229960001419 fenoprofen Drugs 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 229960000389 fluoxetine hydrochloride Drugs 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- XOXYHGOIRWABTC-UHFFFAOYSA-N gentisin Chemical compound C1=C(O)C=C2C(=O)C3=C(O)C=C(OC)C=C3OC2=C1 XOXYHGOIRWABTC-UHFFFAOYSA-N 0.000 description 1
- 229960002972 glutethimide Drugs 0.000 description 1
- 230000010224 hepatic metabolism Effects 0.000 description 1
- LLPOLZWFYMWNKH-CMKMFDCUSA-N hydrocodone Chemical compound C([C@H]1[C@H](N(CC[C@@]112)C)C3)CC(=O)[C@@H]1OC1=C2C3=CC=C1OC LLPOLZWFYMWNKH-CMKMFDCUSA-N 0.000 description 1
- 229960000240 hydrocodone Drugs 0.000 description 1
- OROGSEYTTFOCAN-UHFFFAOYSA-N hydrocodone Natural products C1C(N(CCC234)C)C2C=CC(O)C3OC2=C4C1=CC=C2OC OROGSEYTTFOCAN-UHFFFAOYSA-N 0.000 description 1
- 229960000890 hydrocortisone Drugs 0.000 description 1
- 238000005984 hydrogenation reaction Methods 0.000 description 1
- BCGWQEUPMDMJNV-UHFFFAOYSA-N imipramine Chemical compound C1CC2=CC=CC=C2N(CCCN(C)C)C2=CC=CC=C21 BCGWQEUPMDMJNV-UHFFFAOYSA-N 0.000 description 1
- 229960004801 imipramine Drugs 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 229940095990 inderal Drugs 0.000 description 1
- 230000000053 inderal effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- JEIPFZHSYJVQDO-UHFFFAOYSA-N iron(III) oxide Inorganic materials O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
- 229960004819 isoxsuprine Drugs 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- RDOIQAHITMMDAJ-UHFFFAOYSA-N loperamide Chemical compound C=1C=CC=CC=1C(C=1C=CC=CC=1)(C(=O)N(C)C)CCN(CC1)CCC1(O)C1=CC=C(Cl)C=C1 RDOIQAHITMMDAJ-UHFFFAOYSA-N 0.000 description 1
- 229960001571 loperamide Drugs 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000010297 mechanical methods and process Methods 0.000 description 1
- 230000005226 mechanical processes and functions Effects 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- MYWUZJCMWCOHBA-VIFPVBQESA-N methamphetamine Chemical compound CN[C@@H](C)CC1=CC=CC=C1 MYWUZJCMWCOHBA-VIFPVBQESA-N 0.000 description 1
- 229960001252 methamphetamine Drugs 0.000 description 1
- CRVGTESFCCXCTH-UHFFFAOYSA-N methyl diethanolamine Chemical compound OCCN(C)CCO CRVGTESFCCXCTH-UHFFFAOYSA-N 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- NETZHAKZCGBWSS-CEDHKZHLSA-N nalbuphine Chemical compound C([C@]12[C@H]3OC=4C(O)=CC=C(C2=4)C[C@@H]2[C@]1(O)CC[C@@H]3O)CN2CC1CCC1 NETZHAKZCGBWSS-CEDHKZHLSA-N 0.000 description 1
- 229960000805 nalbuphine Drugs 0.000 description 1
- UZHSEJADLWPNLE-GRGSLBFTSA-N naloxone Chemical compound O=C([C@@H]1O2)CC[C@@]3(O)[C@H]4CC5=CC=C(O)C2=C5[C@@]13CCN4CC=C UZHSEJADLWPNLE-GRGSLBFTSA-N 0.000 description 1
- 229960004127 naloxone Drugs 0.000 description 1
- 229960002009 naproxen Drugs 0.000 description 1
- CMWTZPSULFXXJA-VIFPVBQESA-M naproxen(1-) Chemical compound C1=C([C@H](C)C([O-])=O)C=CC2=CC(OC)=CC=C21 CMWTZPSULFXXJA-VIFPVBQESA-M 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 229950006134 normorphine Drugs 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 229960000988 nystatin Drugs 0.000 description 1
- VQOXZBDYSJBXMA-NQTDYLQESA-N nystatin A1 Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/CC/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 VQOXZBDYSJBXMA-NQTDYLQESA-N 0.000 description 1
- 238000011017 operating method Methods 0.000 description 1
- 229960002085 oxycodone Drugs 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229960000482 pethidine Drugs 0.000 description 1
- 229960003562 phentermine Drugs 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- DLNKOYKMWOXYQA-APPZFPTMSA-N phenylpropanolamine Chemical compound C[C@@H](N)[C@H](O)C1=CC=CC=C1 DLNKOYKMWOXYQA-APPZFPTMSA-N 0.000 description 1
- 229960000395 phenylpropanolamine Drugs 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 229950009829 prasterone sulfate Drugs 0.000 description 1
- 229960000249 pregnenolone Drugs 0.000 description 1
- ORNBQBCIOKFOEO-QGVNFLHTSA-N pregnenolone Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 ORNBQBCIOKFOEO-QGVNFLHTSA-N 0.000 description 1
- REQCZEXYDRLIBE-UHFFFAOYSA-N procainamide Chemical compound CCN(CC)CCNC(=O)C1=CC=C(N)C=C1 REQCZEXYDRLIBE-UHFFFAOYSA-N 0.000 description 1
- 229960000244 procainamide Drugs 0.000 description 1
- MFDFERRIHVXMIY-UHFFFAOYSA-N procaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 MFDFERRIHVXMIY-UHFFFAOYSA-N 0.000 description 1
- 229960004919 procaine Drugs 0.000 description 1
- 239000000186 progesterone Substances 0.000 description 1
- 229960003387 progesterone Drugs 0.000 description 1
- AQHHHDLHHXJYJD-UHFFFAOYSA-N propranolol Chemical compound C1=CC=C2C(OCC(O)CNC(C)C)=CC=CC2=C1 AQHHHDLHHXJYJD-UHFFFAOYSA-N 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- VMXUWOKSQNHOCA-LCYFTJDESA-N ranitidine Chemical compound [O-][N+](=O)/C=C(/NC)NCCSCC1=CC=C(CN(C)C)O1 VMXUWOKSQNHOCA-LCYFTJDESA-N 0.000 description 1
- 229960000620 ranitidine Drugs 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229960002052 salbutamol Drugs 0.000 description 1
- 230000036280 sedation Effects 0.000 description 1
- 229940126570 serotonin reuptake inhibitor Drugs 0.000 description 1
- 239000003772 serotonin uptake inhibitor Substances 0.000 description 1
- 229940124547 specific antidotes Drugs 0.000 description 1
- 239000001384 succinic acid Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000006794 tachycardia Effects 0.000 description 1
- 229960003604 testosterone Drugs 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- LLPOLZWFYMWNKH-UHFFFAOYSA-N trans-dihydrocodeinone Natural products C1C(N(CCC234)C)C2CCC(=O)C3OC2=C4C1=CC=C2OC LLPOLZWFYMWNKH-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/44—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members
- C07D207/444—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5
- C07D207/448—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. maleimide
- C07D207/452—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having three double bonds between ring members or between ring members and non-ring members having two doubly-bound oxygen atoms directly attached in positions 2 and 5 with only hydrogen atoms or radicals containing only hydrogen and carbon atoms directly attached to other ring carbon atoms, e.g. maleimide with hydrocarbon radicals, substituted by hetero atoms, directly attached to the ring nitrogen atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/77—Ovalbumin
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/795—Porphyrin- or corrin-ring-containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/44—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N2035/00465—Separating and mixing arrangements
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Physics & Mathematics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明涉及一种舍曲林检测试剂及其制备和使用方法。通过反复试验获得了一种全新的舍曲林马来酰亚胺衍生物,并使用该舍曲林马来酰亚胺衍生物制备得到高免疫原性的舍曲林免疫原,进而免疫实验动物得到高效价的抗舍曲林特异性抗体;同时,使用该舍曲林马来酰亚胺衍生物制备得到舍曲林酶标偶联物。含有上述抗舍曲林特异性抗体与舍曲林酶标偶联物的舍曲林检测试剂可以实现在全自动生化分析仪上对舍曲林含量的自动化测定,可以高通量、快速化、精确地测定生物样本中舍曲林的含量,且具有操作简便、灵敏度高、特异性强、结果准确等优点,有效降低舍曲林检测成本,有利于临床广泛推广使用。
Description
技术领域
本发明属于生物检测技术领域,具体涉及一种舍曲林检测试剂及其制备和使用方法。
背景技术
舍曲林(Sertraline),其化学结构式如式(Ⅳ)所示:
式(Ⅳ)
舍曲林是一种强效和选择性的神经元5-羟色胺再摄取抑制剂,在临床剂量下,舍曲林阻断人体血小板对5-羟色胺的摄取,对去甲肾上腺素和多巴胺仅有微弱影响。体外研究显示,舍曲林对肾上腺素能受体(α1,α2,β)、胆碱能受体、GABA 受体、多巴胺能受体、组胺受体、5-羟色胺能受体(5HT1A, 5HT1B, 5HT2)或苯二氮卓受体没有明显的亲和力。对上述受体的拮抗作用被认为与其它精神疾病用药的镇静作用、抗胆碱作用和心脏毒性相关。舍曲林对单胺氧化酶没有抑制作用。舍曲林的血浆蛋白结合率为98%,主要通过肝脏代谢。舍曲林用药过量症状包括因5-羟色胺引起的不良反应,如嗜睡、胃肠不适、心动过速、震颤、激动和头晕等,舍曲林没有特效的解毒剂。因此,应对任何服用舍曲林的患者进行药物浓度监测,并对用药过量的患者给予积极治疗。
目前,检测舍曲林的常用方法为气相色谱-质谱法(GC-MS)、高效液相色谱法(HPLC)和酶联免疫分析法(ELISA)等,但是这些方法操作复杂,在临床大规模推广应用方面均具有一定的局限性。目前市场上缺乏稳定性好、灵敏度高、特异性强的舍曲林检测试剂,尤其是质量好的高通量自动化检验试剂。因此,研发一种质量达到临床要求、实用性强、性价比高,可应用于全自动生化分析仪的舍曲林检测试剂已成为国内外体外诊断试剂行业的热点。本发明可以实现在全自动生化分析仪上对舍曲林的高通量、快速化检测,且具有操作简便、灵敏度高、特异性强、结果准确等优点,能有效满足国内日益增长的临床检测需求。
发明内容
本发明要解决的技术问题是:针对现有技术的上述缺陷,提供一种操作简便、灵敏度高、特异性强的舍曲林检测试剂及其制备和使用方法。应用该检测试剂可以实现在全自动生化分析仪上对舍曲林含量的自动化测定,可以高通量、快速化、精确地测定生物样本中舍曲林的含量,且具有操作简便、灵敏度高、特异性强、结果准确等优点,有效降低舍曲林检测成本,有利于临床广泛推广使用。
本发明解决其技术问题所采用的技术方案是:提供一种舍曲林检测试剂,包括试剂R1和试剂R2,所述试剂R1中包含抗舍曲林特异性抗体和均相酶底物溶液;所述试剂R2包含舍曲林酶标偶联物和R2缓冲液。
本发明所述的舍曲林检测试剂,制备方法包括以下步骤:
A.试剂R1的制备:将质量分数3.5%的氧化态的烟酰胺腺嘌呤二核苷酸、3.5%的葡萄糖-6-磷酸、0.12%的甲基化牛血清白蛋白、0.03%的NaN3的用75 mmol/L、pH=7.8的Tris缓冲液溶解制成均相酶底物溶液;再将抗舍曲林特异性抗体加入所述均相酶底物溶液中混匀,得到试剂R1,所述抗舍曲林特异性抗体与均相酶底物溶液的体积比为1:100~1:10000;优选地,所述抗舍曲林特异性抗体与均相酶底物溶液的体积比为1:900。
B.试剂R2的制备:将质量分数0.12%的甲基化牛血清白蛋白、0.03%的NaN3用150mmol/L、pH=8.5的Tris缓冲液溶解制成R2缓冲液,再将舍曲林酶标偶联物加入所述R2缓冲液中混匀,得到试剂R2,所述舍曲林酶标偶联物与R2缓冲液的体积比为1:100~1:10000;优选地,所述舍曲林酶标偶联物与R2缓冲液的体积比为1:3600。
本发明所述的舍曲林均相酶免疫检测试剂,所述的抗舍曲林特异性抗体,由舍曲林免疫原免疫实验动物后产生,所述抗体为完整抗体分子,或者为保留与舍曲林特异性结合能力的抗体片段或抗体衍生物,所述实验动物为山羊、绵羊、小鼠、豚鼠、兔或马中的一种,优选为山羊。
所述抗舍曲林特异性抗体的制备方法,包括以下步骤:
a.用PBS缓冲液将舍曲林免疫原稀释至2.5 mg/ml,得到免疫原溶液,然后用2.5 ml所述免疫原溶液与等量弗氏完全佐剂混合,对实验动物进行注射;
b.2周后,再用2.5 ml相同的免疫原溶液与等量弗氏不完全佐剂混合,对上述实验动物注射一次,之后每隔4周注射一次,共计注射5次;
c.对免疫后的实验动物取血,分离纯化得到抗舍曲林特异性抗体。
本发明所述的舍曲林均相酶免疫检测试剂,其中,所述舍曲林免疫原由舍曲林马来酰亚胺衍生物与载体连接而成,其结构式如下述式(Ⅰ)所示:
式(Ⅰ);
所述载体为具有免疫原性的甲基化蛋白质或多肽,选自甲基化血清蛋白、甲基化卵清蛋白、甲基化血蓝蛋白或甲基化甲状腺球蛋白中的一种,优选为甲基化血清蛋白,更优选为甲基化牛血清白蛋白。
所述舍曲林免疫原的制备方法,包括以下步骤:
(1)将质量分数1.25%的载体蛋白溶解于0.3 mmol/L,pH=8.0的磷酸缓冲液中,得到载体蛋白溶液;
(2)将质量分数0.8%的舍曲林马来酰亚胺衍生物、7.5%的二甲基甲酰胺、7.5%的乙醇溶解于20 mmol/L,pH=5.2的磷酸钾缓冲液中,再加入质量分数0.8%的1-乙基-3-(-3-二甲氨丙基)碳二亚胺,将上述化学物质在0℃下搅拌反应120分钟;
(3)将步骤(2)中溶解好的溶液缓慢滴加至步骤(1)中的载体蛋白溶液中,并在-8℃下搅拌48小时,得到偶联物溶液,将反应后的偶联物溶液进行透析纯化,纯化后所得溶液即为舍曲林免疫原溶液,在舍曲林免疫原溶液中加入质量分数0.08%的NaN3,于-20℃下储存。
本发明所述的舍曲林均相酶免疫检测试剂,其中,所述的舍曲林酶标偶联物由舍曲林马来酰亚胺衍生物与葡萄糖-6-磷酸脱氢酶连接而成,其结构式如下述式(Ⅱ)所示:
式(Ⅱ)。
所述舍曲林酶标偶联物的制备方法,包括以下步骤:
①葡萄糖-6-磷酸脱氢酶溶液的制备:将质量分数5.0%的规格为300KU的葡萄糖-6-磷酸脱氢酶,室温下溶解于含有0.08 mol/L Tris、6.0 mmol/L MgCl2和3.3 mmol/L NaCl,pH=9.2的溶液中;在此溶液中加入质量分数12.5%的还原态的烟酰胺腺嘌呤二核苷酸、7.5%的葡萄糖-6-磷酸以及1.25%的卡必醇;加热到37℃,再缓慢加入质量分数0.75%的二甲基亚砜,摇匀后静置60秒;
②舍曲林马来酰亚胺衍生物的激活:在无水状态下将质量分数0.8%的舍曲林马来酰亚胺衍生物,溶解于7.5%的二甲基甲酰胺中;将此溶液温度降到-8℃;然后加入2.25%的三丁胺、2.75%的氯甲酸异丁酯、1.75%的碳二亚胺;0.75%的N-羟基硫代琥珀酰亚胺;0℃搅拌120分钟;
③葡萄糖-6-磷酸脱氢酶与舍曲林马来酰亚胺衍生物的连接:将步骤②中激活的温度为0℃的舍曲林马来酰亚胺衍生物溶液逐滴加入到步骤①中溶解的温度为37℃的葡萄糖-6-磷酸脱氢酶溶液中;-4℃搅拌48小时;
④纯化产物:将反应后的连接产物通过G-25葡聚糖凝胶层析柱进行纯化,纯化后所得溶液即为舍曲林酶标偶联物溶液,在舍曲林酶标偶联物溶液中加入质量分数0.8%的BSA和质量分数0.08%的NaN3,于-20℃下储存。
本发明所述的舍曲林均相酶免疫检测试剂,其中,所述的舍曲林马来酰亚胺衍生物,其结构式如下述式(Ⅲ)所示:
式(Ⅲ);
上述式(Ⅲ)所示的舍曲林马来酰亚胺衍生物的合成路线如下:
本发明所述的舍曲林均相酶免疫检测试剂,其中,所述的舍曲林马来酰亚胺衍生物的合成路线,具体制备步骤如下:
(a)化合物2的合成:称取7.8 g化合物1溶解于100 ml三氟乙酸中,然后加入7ml三氟甲磺酸和2.3 g硝酸钾,制成反应混合液,室温下搅拌1.5 小时,在此反应混合液中加入100ml纯化水,然后用100ml乙酸乙酯萃取2次,将萃取得到的有机物用200ml卤水冲洗,用MgSO4干燥后进行过滤,再通过减压的方法除去溶剂,最后将得到的粗产物经硅胶纯化得到化合物2,
。
(b)化合物3的合成:将15 g化合物2溶解于300 mL乙醇中,然后加入21 g铁粉混合,将以上混合物回流加热16小时,而后将反应物冷却并进行过滤,将收集到的滤液浓缩得到粗品,再用SiO2快速色谱分析柱对粗品进行纯化,得到化合物3,
。
(c)化合物4的合成:将13.7 g化合物3加入26 mL甲酸中溶解,然后在0℃下加入由25 mL甲酸和49 mL乙酸酐配制的溶液,将此混合物在室温下搅拌2小时,再加入冰水,并用二氯甲烷进行萃取,萃取得到的有机物用1mol/L的氢氧化钠水溶液冲洗,再用卤水洗涤,用Na2SO4进行干燥,过滤后减压除去溶剂,粗产物用硅胶进行纯化,最后用溶剂浓度梯度为100:0-95:5的二氯甲烷:甲醇溶处理,得到化合物4,
。
(d)化合物5的合成:将10 g化合物4加入285 ml丙酮中溶解,然后加入57 ml浓硫酸和285 ml纯化水组成的混合物,并放入冰水浴中,再加入2.2 g亚硝酸钠,将此混合物在冰水浴中搅拌0.5小时,然后加热至室温,将上述混合物在75℃下加热反应2小时,在冰水浴中冷却,再用高浓度的氢氧化铵水溶液终止反应,将反应混合物用乙酸乙酯萃取3次,萃取得到的有机物用纯化水冲洗,用Na2SO4干燥,过滤后减压除去溶剂,粗产物用硅胶进行纯化,最后用溶剂浓度梯度为100:0的二氯甲烷:甲醇到95:5的二氯甲烷:甲醇处理,得到化合物5,
。
(e)化合物6的合成:将3.5 g化合物5加入50 ml二甲基甲酰胺中溶解,然后在0℃下加入1.8 g 4-溴丁酸甲酯与2.8 g K2CO3,再将此混合物在室温下搅拌12小时,然后通过减压蒸发溶剂,最后用SiO2快速色谱分析柱纯化得到化合物6,
。
(f)舍曲林酸衍生物的合成:将2.6 g化合物6加入50 ml MeOH中溶解,然后加入0.25 g LiOH,再将此混合物在室温下搅拌12小时,然后通过减压蒸发溶剂,最后用SiO2快速色谱分析柱纯化得到舍曲林酸衍生物,
。
(g)舍曲林马来酰亚胺衍生物的合成:将0.8 g舍曲林酸衍生物加入5 ml二甲基甲酰胺中溶解,然后的加入1 ml 二异丙基乙胺,0.9 g HATU和0.3 g化合物7,将此反应混合物在室温下搅拌12小时,反应结束后在混合物中加入20 mL纯化水,然后进行过滤,滤渣采用制备级高效液相色谱进行纯化,得到舍曲林马来酰亚胺衍生物,
。
本发明还提供一种如上所述的舍曲林检测试剂的使用方法,包括以下步骤:
(一)在全自动生化分析仪中加入待测样本、R1试剂,混匀,37℃温育3-5分钟;
(二)加入R2试剂,混匀,37℃恒温5-10分钟后,340 nm波长进行检测,连续监测3分钟内的吸光度变化率,由全自动生化分析仪自动计算待测样本中舍曲林的含量;
所述试剂R1与试剂R2按1:1~4:1的体积比使用,优选按4∶1的体积比使用。
本发明所述的舍曲林检测试剂的使用方法,其中,所述的待测样本为生理样本;优选地,所述的生理样本为血清、血浆、尿液、唾液;更优选地,所述的生理样本为血清或血浆。
本发明的有益效果是:通过反复的试验获得了一种全新的舍曲林马来酰亚胺衍生物,并使用该舍曲林马来酰亚胺衍生物制备得到高免疫原性的舍曲林免疫原,进而免疫实验动物得到高效价的抗舍曲林特异性抗体;同时,使用该舍曲林马来酰亚胺衍生物制备得到舍曲林酶标偶联物。含有上述抗舍曲林特异性抗体与舍曲林酶标偶联物的舍曲林检测试剂可以实现在全自动生化分析仪上对舍曲林高通量、快速化的检测。该检测试剂具有操作简便、灵敏度高、特异性强、结果准确等优点,还能有效降低舍曲林检测成本,有利于临床推广使用。
附图说明
图1是实施例6舍曲林均相酶免疫检测方法的标准曲线;
图2是实施例8舍曲林临床标本均相酶免疫检测与高效液相色谱检测对比数据的散点图。
具体实施方式
下面结合附图以及具体实施方式,对本发明做进一步描述,这些附图均为简化的示意图,仅以示意方式说明本发明的基本结构,因此其仅显示与本发明有关的构成。除非特别指明,以下实施例中所用的试剂、仪器、设备、耗材均可从正规渠道商购买获得。
实施例1.舍曲林马来酰亚胺衍生物的合成
舍曲林马来酰亚胺衍生物的化学结构式如式(Ⅲ)所示:
式(Ⅲ)。
上述舍曲林马来酰亚胺衍生物的合成路线及制备步骤如下:
具体的合成步骤如下:
(a)化合物2的合成:称取7.8 g化合物1溶解于100 ml三氟乙酸中,然后加入7ml三氟甲磺酸和2.3 g硝酸钾,制成反应混合液,室温下搅拌1.5 小时,在此反应混合液中加入100ml纯化水,然后用100ml乙酸乙酯萃取2次,将萃取得到的有机物用200ml卤水冲洗,用MgSO4干燥后进行过滤,再通过减压的方法除去溶剂,最后将得到的粗产物经硅胶纯化得到3.8 g化合物2,产率44%,
。
(b)化合物3的合成:将15 g化合物2溶解于300 mL乙醇中,然后加入21 g铁粉混合,将以上混合物回流加热16小时,而后将反应物冷却并进行过滤,将收集到的滤液浓缩得到粗品,再用SiO2快速色谱分析柱对粗品进行纯化,得到9 g化合物3,产率67%,
。
(c)化合物4的合成:将13.7 g化合物3加入26 mL甲酸中溶解,然后在0℃下加入由25 mL甲酸和49 mL乙酸酐配制的溶液,将此混合物在室温下搅拌2小时,再加入冰水,并用二氯甲烷进行萃取,萃取得到的有机物用1mol/L的氢氧化钠水溶液冲洗,再用卤水洗涤,用Na2SO4进行干燥,过滤后减压除去溶剂,粗产物用硅胶进行纯化,最后用溶剂浓度梯度为100:0-95:5的二氯甲烷:甲醇溶处理,得到11g化合物4,产率74%,
。
(d)化合物5的合成:将10 g化合物4加入285 ml丙酮中溶解,然后加入57 ml浓硫酸和285 ml纯化水组成的混合物,并放入冰水浴中,再加入2.2 g亚硝酸钠,将此混合物在冰水浴中搅拌0.5小时,然后加热至室温,将上述混合物在75℃下加热反应2小时,在冰水浴中冷却,再用高浓度的氢氧化铵水溶液终止反应,将反应混合物用乙酸乙酯萃取3次,萃取得到的有机物用纯化水冲洗,用Na2SO4干燥,过滤后减压除去溶剂,粗产物用硅胶进行纯化,最后用溶剂浓度梯度为100:0的二氯甲烷:甲醇到95:5的二氯甲烷:甲醇处理,得到6.6 g化合物5,产率66%,
。
(e)化合物6的合成:将3.5 g化合物5加入50 ml二甲基甲酰胺中溶解,然后在0℃下加入1.8 g 4-溴丁酸甲酯与2.8 g K2CO3,再将此混合物在室温下搅拌12小时,然后通过减压蒸发溶剂,最后用SiO2快速色谱分析柱纯化得到2.9 g化合物6,产率64%,
。
(f)舍曲林酸衍生物的合成:将2.6 g化合物6加入50 ml MeOH中溶解,然后加入0.25 g LiOH,再将此混合物在室温下搅拌12小时,然后通过减压蒸发溶剂,最后用SiO2快速色谱分析柱纯化得到1.9 g舍曲林酸衍生物,产率80%,
。
(g)舍曲林马来酰亚胺衍生物的合成:将0.8 g舍曲林酸衍生物加入5 ml二甲基甲酰胺中溶解,然后的加入1 ml 二异丙基乙胺,0.9 g HATU和0.3 g化合物7,将此反应混合物在室温下搅拌12小时,反应结束后在混合物中加入20 mL纯化水,然后进行过滤,滤渣采用制备级高效液相色谱进行纯化,得到0.6 g舍曲林马来酰亚胺衍生物,为白色固体,产率55%,
对上述合成步骤得到的舍曲林马来酰亚胺衍生物利用VARIAN MERCURY plus400MHz进行1H核磁共振光谱扫描,采用TMS作为内标进行结构鉴定,并用LC-MS方法检测最终产物纯度,结果显示:纯度>95%。
实施例2.舍曲林免疫原的制备
本实施例中的舍曲林免疫原由式(Ⅲ)所示的舍曲林马来酰亚胺衍生物与甲基化牛血清白蛋白(MBSA)连接而成,其结构式如下述式(Ⅴ)所示:
式(Ⅴ)。
该舍曲林免疫原的合成方法具体步骤如下:
(1)将质量分数1.25%的MBSA溶解于0.3 mmol/L,pH=8.0的磷酸缓冲液中,得到MBSA溶液;
(2)将质量分数0.8%的舍曲林马来酰亚胺衍生物、7.5%的二甲基甲酰胺、7.5%的乙醇溶解于20 mmol/L,pH=5.2的磷酸钾缓冲液中,再加入质量分数0.8%的1-乙基-3-(-3-二甲氨丙基)碳二亚胺,将上述化学物质在0℃下搅拌反应120分钟;
(3)将步骤(2)中溶解好的溶液缓慢滴加至步骤(1)中的MBSA溶液中,并在-8℃下搅拌48小时,得到偶联物溶液,将反应后的偶联物溶液进行透析纯化,纯化后所得溶液即为舍曲林免疫原溶液,在舍曲林免疫原溶液中加入质量分数0.08%的NaN3,于-20℃下储存。
实施例3.抗舍曲林特异性抗体的制备
将实施例2制备得到的舍曲林免疫原采用常规方法接种实验动物山羊,加强免疫后取抗血清,具体步骤如下:
a.用PBS缓冲液将舍曲林免疫原稀释至2.5 mg/ml,得到免疫原溶液,然后用2.5 ml所述免疫原溶液与等量弗氏完全佐剂混合,对实验动物山羊进行注射;
b. 2周后,再用2.5 ml相同的免疫原溶液与等量弗氏不完全佐剂混合,对上述实验动物山羊注射一次,之后每隔4周注射一次,共计注射5次;
c.对免疫后的实验动物山羊取血,分离纯化得到抗舍曲林特异性抗体,测定其效价为1∶12500。
实施例4.舍曲林酶标偶联物的制备
本实施例中的舍曲林酶标偶联物由式(Ⅲ)所示的舍曲林马来酰亚胺衍生物与葡萄糖-6-磷酸脱氢酶(G6PDH)连接而成,其结构式如下述式(Ⅱ)所示:
式(Ⅱ);
该舍曲林酶标偶联物的合成方法具体步骤如下:
①葡萄糖-6-磷酸脱氢酶溶液的制备:将质量分数5.0%的规格为300KU的葡萄糖-6-磷酸脱氢酶,室温下溶解于含有0.08 mol/L Tris、6.0 mmol/L MgCl2和3.3 mmol/L NaCl,pH=9.2的溶液中;在此溶液中加入质量分数12.5%的还原态的烟酰胺腺嘌呤二核苷酸、7.5%的葡萄糖-6-磷酸以及1.25%的卡必醇;加热到37℃,再缓慢加入质量分数0.75%的二甲基亚砜,摇匀后静置60秒;
②舍曲林马来酰亚胺衍生物的激活:在无水状态下将质量分数0.8%的舍曲林马来酰亚胺衍生物,溶解于7.5%的二甲基甲酰胺中;将此溶液温度降到-8℃;然后加入2.25%的三丁胺、2.75%的氯甲酸异丁酯、1.75%的碳二亚胺;0.75%的N-羟基硫代琥珀酰亚胺;0℃搅拌120分钟;
③葡萄糖-6-磷酸脱氢酶与舍曲林马来酰亚胺衍生物的连接:将步骤②中激活的温度为0℃的舍曲林马来酰亚胺衍生物溶液逐滴加入到步骤①中溶解的温度为37℃的葡萄糖-6-磷酸脱氢酶溶液中;-4℃搅拌48小时;
④纯化产物:将反应后的连接产物通过G-25葡聚糖凝胶层析柱进行纯化,纯化后所得溶液即为舍曲林酶标偶联物溶液,在舍曲林酶标偶联物溶液中加入质量分数0.8%的BSA和质量分数0.08%的NaN3,于-20℃下储存。
实施例5.舍曲林均相酶免疫检测试剂的制备
A.试剂R1的制备:将质量分数3.5%的氧化态的烟酰胺腺嘌呤二核苷酸、3.5%的葡萄糖-6-磷酸、0.12%的甲基化牛血清白蛋白、0.03%的 NaN3的用75 mmol/L、pH=7.8的Tris缓冲液溶解制成均相酶底物溶液;再将抗舍曲林特异性抗体加入所述均相酶底物溶液中混匀,得到试剂R1,所述抗舍曲林特异性抗体与均相酶底物溶液的体积比为1: 900;
B.试剂R2的制备:将质量分数0.12%的甲基化牛血清白蛋白、0.03%的NaN3用150 mmol/L、pH=8.5的Tris缓冲液溶解制成R2缓冲液,再将舍曲林酶标偶联物加入所述R2缓冲液中混匀,得到试剂R2,所述舍曲林酶标偶联物与R2缓冲液的体积比为1: 3600。
实施例6. 舍曲林均相酶免疫检验及结果
1. 获得标准曲线:
(1)设置迈瑞 BS480全自动生化分析仪反应参数(表1)。
(2)操作步骤为:先加试剂R1,再加入标准品,最后加入试剂R2。加入试剂R2后,测定不同时间点的OD340吸光值,算出不同标准品浓度时的反应速率,实际操作过程中需不断调整试剂R1和试剂R2的体积比例,同时调整测光点,最后得出较理想的反应标准曲线图,如图1所示。
表1 迈瑞BS480全自动生化分析仪反应参数
2. 样本检测:通过本发明的舍曲林均相酶免疫检测试剂得到的标准曲线,重复测定低、中、高浓度质控样本10次,上述质控样本为:将舍曲林标准品溶解于空白人造血浆中,至浓度分别为3.00,15.00,30.00 ng/ml。检测结果及数据分析见表2。
表2 样本测定值及精密度和回收率评估
血液样本 | 低 | 中 | 高 |
样本浓度(ng/ml) | 3.00 | 15.00 | 30.00 |
1 | 3.14 | 15.47 | 31.19 |
2 | 3.26 | 15.03 | 30.31 |
3 | 2.98 | 15.54 | 30.06 |
4 | 3.15 | 15.28 | 29.87 |
5 | 3.09 | 14.67 | 29.98 |
6 | 3.15 | 15.19 | 30.99 |
7 | 2.97 | 15.32 | 29.45 |
8 | 3.02 | 14.75 | 30.61 |
9 | 3.08 | 15.77 | 30.53 |
10 | 3.21 | 15.50 | 30.24 |
平均值(ng/ml) | 3.11 | 15.25 | 30.32 |
标准差(SD) | 0.95 | 3.51 | 5.24 |
精密度(CV%) | 3.05 | 2.30 | 1.73 |
回收率(%) | 103.67 | 101.67 | 101.07 |
检测结果:本发明的舍曲林均相酶免疫检测试剂测定的准确度高,回收率均在95%-105%之间;精密度高,CV均低于5%。
实施例7:药物与激素干扰试验
选取62种常见药物与30种常见激素及激素代谢物作为干扰物,进行干扰试验,调整浓度至100.0 ng/mL,采用实施例5的舍曲林均相酶免疫检测试剂进行测定:将待测干扰物与实施例5制备的试剂R1接触反应,再加入试剂R2;检测上述混合溶液的OD340吸光值,根据图1的标准曲线得到相应物质的浓度。62种常见药物与30种常见激素及激素代谢物名称以及测定结果详见表3。
表3常见干扰物测定结果
ID# | 化合物名称 | 等价于舍曲林的浓度 (ng/mL) | ID# | 化合物名称 | 等价于舍曲林的浓度(ng/mL) |
1 | 阿司匹林 | 0.00 | 2 | 苯丙醇胺 | 0.00 |
3 | β-苯基乙胺 | 0.00 | 4 | 普鲁卡因酰胺 | 0.00 |
5 | 安非他命 | 0.00 | 6 | 普鲁卡因 | 0.00 |
7 | 氨苄青霉素 | 0.00 | 8 | 奎尼丁 | 0.00 |
9 | 甲氨二氮卓 | 0.00 | 10 | 佐美酸 | 0.00 |
11 | 氯丙嗪 | 0.00 | 12 | 苯肾上腺素 | 0.00 |
13 | 氯拉卓酸 | 0.00 | 14 | 桂皮酰艾克宁 | 0.00 |
15 | 二甲苯氧庚酸 | 0.00 | 16 | 芽子碱 | 0.00 |
17 | 非诺洛芬 | 0.00 | 18 | 地西洋 | 0.00 |
19 | 甲基苯丙胺 | 0.00 | 20 | 可替宁 | 0.00 |
21 | 龙胆酸 | 0.00 | 22 | 阿替洛尔 | 0.00 |
23 | 吉非贝齐 | 0.00 | 24 | 心得安 | 0.00 |
25 | 氢可酮 | 0.00 | 26 | 苯乙哌啶酮 | 0.00 |
27 | 布洛芬 | 0.00 | 28 | 苯基丁氮酮 | 0.00 |
29 | 丙咪嗪 | 0.00 | 30 | 麦角酸二乙基酰胺 | 0.00 |
31 | 二氨基二苯砜 | 0.00 | 32 | 大麻酚 | 0.00 |
33 | 萘普生 | 0.00 | 34 | 洛哌丁胺 | 0.00 |
35 | 氢氯噻嗪 | 0.00 | 36 | 异克舒令 | 0.00 |
37 | 哌替啶 | 0.00 | 38 | 苯基丙氨酸 | 0.00 |
39 | 烯丙羟吗啡酮 | 0.00 | 40 | 盐酸氟西汀 | 0.00 |
41 | 麻黄素 | 0.00 | 42 | 柳丁氨醇 | 0.00 |
43 | 烟酰胺 | 0.00 | 44 | 青霉素 | 0.00 |
45 | 甲胺呋硫 | 0.00 | 46 | 甲基二乙醇胺 | 0.00 |
47 | 异戊巴比妥 | 0.00 | 48 | 二亚甲基双氧苯丙胺 | 0.00 |
49 | 甲撑二氧苯丙胺 | 0.00 | 50 | 琥珀酸多西拉敏 | 0.00 |
51 | 四氢大麻酚 | 0.00 | 52 | 纳布啡 | 0.00 |
53 | 制霉菌素 | 0.00 | 54 | 去甲吗啡 | 0.00 |
55 | 乙酰吗啡 | 0.00 | 56 | 羟考酮 | 0.00 |
57 | 苄非他明 | 0.00 | 58 | 克他命 | 0.00 |
59 | 异丙嗪 | 0.00 | 60 | 苯海拉明 | 0.00 |
61 | 阿司帕坦 | 0.00 | 62 | 苯丁胺 | 0.00 |
63 | 皮质醇 (氢化可的松) | 0.00 | 64 | 醛固酮 | 0.00 |
65 | 雄烯二酮 | 0.00 | 66 | 雄甾酮 | 0.00 |
67 | 皮质脂酮 | 0.00 | 68 | 皮质酮 (可的松) | 0.00 |
69 | 去氧皮质酮 | 0.00 | 70 | 脱氢表雄酮 | 0.00 |
71 | 硫酸脱氢表雄酮 | 0.00 | 72 | 二氢睾酮 | 0.00 |
73 | 雌二醇 | 0.00 | 74 | 雌三醇 | 0.00 |
75 | 雌酮 | 0.00 | 76 | 本胆烷醇酮 | 0.00 |
77 | 17-羟孕烯醇酮 | 0.00 | 78 | 17-羟孕酮 | 0.00 |
79 | 孕烯醇酮 | 0.00 | 80 | 孕酮 | 0.00 |
81 | 睾酮 | 0.00 | 82 | 孕三醇 | 0.00 |
83 | 孕二醇 | 0.00 | 84 | 17α-羟基黄体酮 | 0.00 |
85 | 雄烯二酮 | 0.00 | 86 | 17-酮类固醇 | 0.00 |
87 | 17-羟皮质类固醇 | 0.00 | 88 | 肾上腺素 | 0.00 |
89 | 去甲肾上腺素 | 0.00 | 90 | 多巴胺 | 0.00 |
91 | 高香草酸 | 0.00 | 92 | 二羟基杏仁酸 | 0.00 |
测定结果显示:上述62种常见药物与30种常见激素及激素代谢物等价于舍曲林的浓度均小于1.00 ng/mL。由此可见,本发明的抗体是抗舍曲林的特异性抗体,与常见干扰物无交叉反应。
实施例8:相关性分析
对100例临床标本分别使用高效液相色谱法和实施例5的舍曲林均相酶免疫检测试剂进行测定,并作相关性分析,测定的数据详见表4。
表4 临床标本测定结果对比数据
样本号 | 均相酶免疫检测试剂测定值(ng/mL) | 高效液相色谱法测定值(ng/mL) |
1 | 14.11 | 14.32 |
2 | 15.98 | 16.09 |
3 | 5.49 | 5.32 |
4 | 2.32 | 2.38 |
5 | 13.23 | 13.35 |
6 | 11.02 | 11.06 |
7 | 14.85 | 14.66 |
8 | 17.85 | 17.94 |
9 | 7.13 | 7.24 |
10 | 2.04 | 2.05 |
11 | 5.95 | 6.08 |
12 | 5.10 | 5.23 |
13 | 10.03 | 10.17 |
14 | 2.25 | 2.36 |
15 | 3.24 | 3.36 |
16 | 15.3 | 15.26 |
17 | 11.81 | 11.98 |
18 | 14.58 | 14.69 |
19 | 14.54 | 14.33 |
20 | 3.98 | 4.03 |
21 | 15.01 | 15.21 |
22 | 18.17 | 18.25 |
23 | 16.05 | 16.22 |
24 | 14.26 | 14.25 |
25 | 17.21 | 17.55 |
26 | 0.36 | 0.33 |
27 | 4.98 | 5.03 |
28 | 1.99 | 2.02 |
29 | 16.01 | 16.23 |
30 | 15.05 | 15.25 |
31 | 17.85 | 17.89 |
32 | 17.15 | 17.23 |
33 | 11.41 | 11.22 |
34 | 10.59 | 10.55 |
35 | 18.03 | 18.25 |
36 | 13.96 | 14.06 |
37 | 18.56 | 18.69 |
38 | 17.15 | 17.22 |
39 | 16.04 | 16.23 |
40 | 13.09 | 13.12 |
41 | 2.58 | 2.56 |
42 | 5.87 | 6.01 |
43 | 1.15 | 1.18 |
44 | 1.20 | 1.23 |
45 | 5.19 | 5.23 |
46 | 2.35 | 2.36 |
47 | 5.21 | 5.23 |
48 | 18.59 | 18.79 |
49 | 5.31 | 5.22 |
50 | 17.03 | 17.25 |
51 | 9.56 | 9.69 |
52 | 5.02 | 5.02 |
53 | 12.69 | 12.87 |
54 | 14.16 | 14.35 |
55 | 17.01 | 17.22 |
56 | 2.06 | 2.03 |
57 | 3.99 | 4.03 |
58 | 1.18 | 1.22 |
59 | 11.35 | 11.96 |
60 | 10.02 | 10.39 |
61 | 13.88 | 14.21 |
62 | 11.23 | 11.56 |
63 | 2.01 | 2.03 |
64 | 15.21 | 15.36 |
65 | 8.27 | 8.03 |
66 | 18.05 | 17.96 |
67 | 12.49 | 12.33 |
68 | 16.34 | 16.13 |
69 | 4.03 | 4.28 |
70 | 5.88 | 6.01 |
71 | 2.26 | 2.23 |
72 | 4.52 | 4.47 |
73 | 15.69 | 15.38 |
74 | 17.15 | 17.29 |
75 | 14.41 | 14.35 |
76 | 10.96 | 10.99 |
77 | 16.23 | 16.33 |
78 | 15.86 | 15.79 |
79 | 3.41 | 3.22 |
80 | 4.21 | 4.05 |
81 | 5.46 | 5.55 |
82 | 17.91 | 17.58 |
83 | 10.76 | 10.99 |
84 | 17.89 | 17.68 |
85 | 2.42 | 2.35 |
86 | 4.02 | 4.22 |
87 | 8.79 | 8.99 |
88 | 15.48 | 15.23 |
89 | 4.42 | 4.39 |
90 | 8.49 | 8.56 |
91 | 7.36 | 7.22 |
92 | 5.16 | 5.23 |
93 | 8.67 | 8.97 |
94 | 1.96 | 1.88 |
95 | 12.34 | 12.58 |
96 | 2.45 | 2.55 |
97 | 6.09 | 6.34 |
98 | 15.62 | 15.79 |
99 | 17.52 | 17.69 |
100 | 2.38 | 2.33 |
对上述数据作图,参见图2,得到的线性方程为:y = 1.0034x + 0.0271,相关系数R2 =0.9993,表明本发明的舍曲林检测试剂测定舍曲林临床标本的准确度高,与高效液相色谱法的一致性较好。
以上述依据本发明的理想实施例为启示,通过上述的说明内容,相关技术人员完全可以在不偏离本项发明技术思想的范围内,进行多样的变更以及修改。本项发明的技术性范围并不局限于说明书上的内容,必须要根据权利要求范围来确定其技术性范围。
Claims (9)
1.一种舍曲林检测试剂,其特征在于,包括试剂R1和试剂R2,所述试剂R1中包含抗舍曲林特异性抗体和均相酶底物溶液;所述试剂R2包含舍曲林酶标偶联物和R2缓冲液。
2.根据权利要求1所述的舍曲林检测试剂,其特征在于,制备方法包括以下步骤:
A.试剂R1的制备:将质量分数3.5%的氧化态的烟酰胺腺嘌呤二核苷酸、3.5%的葡萄糖-6-磷酸、0.12%的甲基化牛血清白蛋白、0.03%的NaN3的用75 mmol/L、pH=7.8的Tris缓冲液溶解制成均相酶底物溶液;再将抗舍曲林特异性抗体加入所述均相酶底物溶液中混匀,得到试剂R1,所述抗舍曲林特异性抗体与均相酶底物溶液的体积比为1:100~1:10000;优选地,所述抗舍曲林特异性抗体与均相酶底物溶液的体积比为1:900;
B.试剂R2的制备:将质量分数0.12%的甲基化牛血清白蛋白、0.03%的NaN3用150 mmol/L、pH=8.5的Tris缓冲液溶解制成R2缓冲液,再将舍曲林酶标偶联物加入所述R2缓冲液中混匀,得到试剂R2,所述舍曲林酶标偶联物与R2缓冲液的体积比为1:100~1:10000;优选地,所述舍曲林酶标偶联物与R2缓冲液的体积比为1:3600。
3.根据权利要求1-2中任一项所述的舍曲林均相酶免疫检测试剂,其特征在于,所述的抗舍曲林特异性抗体,由舍曲林免疫原免疫实验动物后产生,所述抗体为完整抗体分子,或者为保留与舍曲林特异性结合能力的抗体片段或抗体衍生物,所述实验动物为山羊、绵羊、小鼠、豚鼠、兔或马中的一种,优选为山羊;
所述抗舍曲林特异性抗体的制备方法,包括以下步骤:
a.用PBS缓冲液将舍曲林免疫原稀释至2.5 mg/ml,得到免疫原溶液,然后用2.5 ml所述免疫原溶液与等量弗氏完全佐剂混合,对实验动物进行注射;
b.2周后,再用2.5 ml相同的免疫原溶液与等量弗氏不完全佐剂混合,对上述实验动物注射一次,之后每隔4周注射一次,共计注射5次;
c.对免疫后的实验动物取血,分离纯化得到抗舍曲林特异性抗体。
4.根据权利要求3所述的舍曲林均相酶免疫检测试剂,其特征在于,所述舍曲林免疫原由舍曲林马来酰亚胺衍生物与载体连接而成,其结构式如下述式(Ⅰ)所示:
式(Ⅰ);
所述载体为具有免疫原性的甲基化蛋白质或多肽,选自甲基化血清蛋白、甲基化卵清蛋白、甲基化血蓝蛋白或甲基化甲状腺球蛋白中的一种,优选为甲基化血清蛋白,更优选为甲基化牛血清白蛋白;
所述舍曲林免疫原的制备方法,包括以下步骤:
(1)将质量分数1.25%的载体蛋白溶解于0.3 mmol/L,pH=8.0的磷酸缓冲液中,得到载体蛋白溶液;
(2)将质量分数0.8%的舍曲林马来酰亚胺衍生物、7.5%的二甲基甲酰胺、7.5%的乙醇溶解于20 mmol/L,pH=5.2的磷酸钾缓冲液中,再加入质量分数0.8%的1-乙基-3-(-3-二甲氨丙基)碳二亚胺,将上述化学物质在0℃下搅拌反应120分钟;
(3)将步骤(2)中溶解好的溶液缓慢滴加至步骤(1)中的载体蛋白溶液中,并在-8℃下搅拌48小时,得到偶联物溶液,将反应后的偶联物溶液进行透析纯化,纯化后所得溶液即为舍曲林免疫原溶液,在舍曲林免疫原溶液中加入质量分数0.08%的NaN3,于-20℃下储存。
5.根据权利要求1-2中任一项所述的舍曲林均相酶免疫检测试剂,其特征在于,所述的舍曲林酶标偶联物由舍曲林马来酰亚胺衍生物与葡萄糖-6-磷酸脱氢酶连接而成,其结构式如下述式(Ⅱ)所示:
式(Ⅱ);
所述舍曲林酶标偶联物的制备方法,包括以下步骤:
①葡萄糖-6-磷酸脱氢酶溶液的制备:将质量分数5.0%的规格为300KU的葡萄糖-6-磷酸脱氢酶,室温下溶解于含有0.08 mol/L Tris、6.0 mmol/L MgCl2和3.3 mmol/L NaCl,pH=9.2的溶液中;在此溶液中加入质量分数12.5%的还原态的烟酰胺腺嘌呤二核苷酸、7.5%的葡萄糖-6-磷酸以及1.25%的卡必醇;加热到37℃,再缓慢加入质量分数0.75%的二甲基亚砜,摇匀后静置60秒;
②舍曲林马来酰亚胺衍生物的激活:在无水状态下将质量分数0.8%的舍曲林马来酰亚胺衍生物,溶解于7.5%的二甲基甲酰胺中;将此溶液温度降到-8℃;然后加入2.25%的三丁胺、2.75%的氯甲酸异丁酯、1.75%的碳二亚胺;0.75%的N-羟基硫代琥珀酰亚胺;0℃搅拌120分钟;
③葡萄糖-6-磷酸脱氢酶与舍曲林马来酰亚胺衍生物的连接:将步骤②中激活的温度为0℃的舍曲林马来酰亚胺衍生物溶液逐滴加入到步骤①中溶解的温度为37℃的葡萄糖-6-磷酸脱氢酶溶液中;-4℃搅拌48小时;
④纯化产物:将反应后的连接产物通过G-25葡聚糖凝胶层析柱进行纯化,纯化后所得溶液即为舍曲林酶标偶联物溶液,在舍曲林酶标偶联物溶液中加入质量分数0.8%的BSA和质量分数0.08%的NaN3,于-20℃下储存。
6.根据权利要求4-5中任一项所述的舍曲林均相酶免疫检测试剂,其特征在于,所述的舍曲林马来酰亚胺衍生物,其结构式如下述式(Ⅲ)所示:
式(Ⅲ);
上述式(Ⅲ)所示的舍曲林马来酰亚胺衍生物的合成路线如下:
。
7.根据权利要求6所述的舍曲林均相酶免疫检测试剂,其特征在于,所述的舍曲林马来酰亚胺衍生物的合成路线,具体制备步骤如下:
(a)化合物2的合成:称取7.8 g化合物1溶解于100 ml三氟乙酸中,然后加入7ml三氟甲磺酸和2.3 g硝酸钾,制成反应混合液,室温下搅拌1.5 小时,在此反应混合液中加入100ml纯化水,然后用100ml乙酸乙酯萃取2次,将萃取得到的有机物用200ml卤水冲洗,用MgSO4干燥后进行过滤,再通过减压的方法除去溶剂,最后将得到的粗产物经硅胶纯化得到化合物2,
;
(b)化合物3的合成:将15 g化合物2溶解于300 mL乙醇中,然后加入21 g铁粉混合,将以上混合物回流加热16小时,而后将反应物冷却并进行过滤,将收集到的滤液浓缩得到粗品,再用SiO2快速色谱分析柱对粗品进行纯化,得到化合物3,
;
(c)化合物4的合成:将13.7 g化合物3加入26 mL甲酸中溶解,然后在0℃下加入由25mL甲酸和49 mL乙酸酐配制的溶液,将此混合物在室温下搅拌2小时,再加入冰水,并用二氯甲烷进行萃取,萃取得到的有机物用1mol/L的氢氧化钠水溶液冲洗,再用卤水洗涤,用Na2SO4进行干燥,过滤后减压除去溶剂,粗产物用硅胶进行纯化,最后用溶剂浓度梯度为100:0-95:5的二氯甲烷:甲醇溶处理,得到化合物4,
;
(d)化合物5的合成:将10 g化合物4加入285 ml丙酮中溶解,然后加入57 ml浓硫酸和285 ml纯化水组成的混合物,并放入冰水浴中,再加入2.2 g亚硝酸钠,将此混合物在冰水浴中搅拌0.5小时,然后加热至室温,将上述混合物在75℃下加热反应2小时,在冰水浴中冷却,再用高浓度的氢氧化铵水溶液终止反应,将反应混合物用乙酸乙酯萃取3次,萃取得到的有机物用纯化水冲洗,用Na2SO4干燥,过滤后减压除去溶剂,粗产物用硅胶进行纯化,最后用溶剂浓度梯度为100:0的二氯甲烷:甲醇到95:5的二氯甲烷:甲醇处理,得到化合物5,
;
(e)化合物6的合成:将3.5 g化合物5加入50 ml二甲基甲酰胺中溶解,然后在0℃下加入1.8 g 4-溴丁酸甲酯与2.8 g K2CO3,再将此混合物在室温下搅拌12小时,然后通过减压蒸发溶剂,最后用SiO2快速色谱分析柱纯化得到化合物6,
;
(f)舍曲林酸衍生物的合成:将2.6 g化合物6加入50 ml MeOH中溶解,然后加入0.25g LiOH,再将此混合物在室温下搅拌12小时,然后通过减压蒸发溶剂,最后用SiO2快速色谱分析柱纯化得到舍曲林酸衍生物,
;
(g)舍曲林马来酰亚胺衍生物的合成:将0.8 g舍曲林酸衍生物加入5 ml二甲基甲酰胺中溶解,然后的加入1 ml 二异丙基乙胺,0.9 g HATU和0.3 g化合物7,将此反应混合物在室温下搅拌12小时,反应结束后在混合物中加入20 mL纯化水,然后进行过滤,滤渣采用制备级高效液相色谱进行纯化,得到舍曲林马来酰亚胺衍生物,
。
8.一种如权利要求1所述的舍曲林检测试剂的使用方法,其特征在于,包括以下步骤:
(一)在全自动生化分析仪中加入待测样本、R1试剂,混匀,37℃温育3-5分钟;
(二)加入R2试剂,混匀,37℃恒温5-10分钟后,340 nm波长进行检测,连续监测3分钟内的吸光度变化率,由全自动生化分析仪自动计算待测样本中舍曲林的含量;
所述试剂R1与试剂R2按1:1~4:1的体积比使用,优选按4:1的体积比使用。
9.根据权利要求8所述的舍曲林检测试剂的使用方法,其特征在于,所述的待测样本为生理样本;优选地,所述的生理样本为血清、血浆、尿液、唾液;更优选地,所述的生理样本为血清或血浆。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910767565.5A CN110456087B (zh) | 2019-08-20 | 2019-08-20 | 一种舍曲林检测试剂及其制备和使用方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201910767565.5A CN110456087B (zh) | 2019-08-20 | 2019-08-20 | 一种舍曲林检测试剂及其制备和使用方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110456087A true CN110456087A (zh) | 2019-11-15 |
CN110456087B CN110456087B (zh) | 2022-09-20 |
Family
ID=68487926
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201910767565.5A Active CN110456087B (zh) | 2019-08-20 | 2019-08-20 | 一种舍曲林检测试剂及其制备和使用方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN110456087B (zh) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111018924A (zh) * | 2019-12-19 | 2020-04-17 | 苏州博源医疗科技有限公司 | 一种柔红霉素衍生物及其制备方法与柔红霉素检测试剂 |
CN111875587B (zh) * | 2020-07-23 | 2021-07-13 | 湖南苏阳医疗科技有限公司 | 5-氟胞嘧啶衍生物、其制备方法及其在5-氟胞嘧啶免疫检测试剂中的应用 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109085264A (zh) * | 2018-08-03 | 2018-12-25 | 杭州佰勤医疗器械有限公司 | 液相色谱串联质谱法检测血清血浆中抗抑郁药物的试剂盒及其应用 |
CN110455945A (zh) * | 2019-08-06 | 2019-11-15 | 北京回龙观医院(北京心理危机研究与干预中心) | 一种检测血液中5种精神药物及其主要代谢产物的方法及试剂盒 |
-
2019
- 2019-08-20 CN CN201910767565.5A patent/CN110456087B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN109085264A (zh) * | 2018-08-03 | 2018-12-25 | 杭州佰勤医疗器械有限公司 | 液相色谱串联质谱法检测血清血浆中抗抑郁药物的试剂盒及其应用 |
CN110455945A (zh) * | 2019-08-06 | 2019-11-15 | 北京回龙观医院(北京心理危机研究与干预中心) | 一种检测血液中5种精神药物及其主要代谢产物的方法及试剂盒 |
Non-Patent Citations (2)
Title |
---|
BRENDA B.SUH-LAILAM等: "Performance characteristics of three assays for the therapeutic drug monitoring of methotrexate", 《THER DRUG MONIT》 * |
齐谢敏等: "均相酶免疫分析技术在治疗药物监测中的应用", 《药学与临床研究》 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111018924A (zh) * | 2019-12-19 | 2020-04-17 | 苏州博源医疗科技有限公司 | 一种柔红霉素衍生物及其制备方法与柔红霉素检测试剂 |
CN111018924B (zh) * | 2019-12-19 | 2022-12-27 | 苏州博源医疗科技有限公司 | 一种柔红霉素衍生物及其制备方法与柔红霉素检测试剂 |
CN111875587B (zh) * | 2020-07-23 | 2021-07-13 | 湖南苏阳医疗科技有限公司 | 5-氟胞嘧啶衍生物、其制备方法及其在5-氟胞嘧啶免疫检测试剂中的应用 |
Also Published As
Publication number | Publication date |
---|---|
CN110456087B (zh) | 2022-09-20 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN106645692B (zh) | 雌三醇均相酶免疫检测试剂、制备方法及检测方法 | |
US9720004B2 (en) | Immunoassays employing non-particulate chemiluminescent reagent | |
US20070254323A1 (en) | Malachite green derivatives for immunoassay reagents to detect malachite green | |
CN103575890B (zh) | 一种莱克多巴胺的化学发光试剂盒及其应用 | |
CN109111494A (zh) | 雌二醇衍生物、免疫原、抗体、酶标偶联物、检测试剂及其制备方法 | |
CN102422160B (zh) | 利用免疫法测定生物样品中的雌马酚的方法、用于该测定的试剂盒、以及判定受试者的雌马酚产生能力的方法 | |
Li et al. | Homogeneous substrate-labeled fluorescent immunoassay for theophylline in serum. | |
CN104569373B (zh) | 一种甲氨蝶呤均相酶免疫检测试剂及其制备和检测方法 | |
CN104788560B (zh) | 环孢霉素a免疫原、抗环孢霉素a特异性抗体和环孢霉素a检测试剂 | |
JPS6214064A (ja) | 置換カルボキシフルオレセイン | |
CN104447984A (zh) | 多西紫杉醇免疫原、抗多西紫杉醇特异性抗体和多西紫杉醇检测试剂 | |
CN110456087A (zh) | 一种舍曲林检测试剂及其制备和使用方法 | |
CN109917131A (zh) | 一种脂蛋白磷脂酶a2检测试剂及其制备和使用方法 | |
CN105175530A (zh) | 一种香草扁桃酸免疫检测试剂及其制备方法 | |
CN105175531A (zh) | 羟脯氨酸免疫原、特异性抗体、检测试剂及其制备方法 | |
CN110003300A (zh) | 一种17-羟类固醇的衍生物、检测试剂及制备方法 | |
CN104804079A (zh) | 伊马替尼免疫原、衍生物及合成方法、特异性抗体和检测试剂及制备方法 | |
CN104447745A (zh) | 一种茶碱均相酶免疫检测验试剂盒及其制备方法 | |
CN106596917B (zh) | 高香草酸均相酶免疫检测试剂、制备方法及检测方法 | |
CN105131105A (zh) | 皮质醇免疫原、衍生物、抗体、检测试剂及制备方法 | |
CN106645764B (zh) | 检测地西泮的酶联免疫试剂盒及其应用 | |
CN107132349A (zh) | 一种同型半胱氨酸自动化检测试剂、制备方法及检测方法 | |
CN108586562B (zh) | 一种皮质醇衍生物及其制备方法与应用 | |
CN105061339A (zh) | 一种半抗原、其人工抗原及其在检测喹乙醇残留标志物中的应用 | |
CN105801688A (zh) | 脱氧吡啶酚免疫原、抗体和检测试剂及制备方法 |
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 | ||
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20230614 Address after: Room 302, 3 / F, building A5, phase II, Changsha E center, No.18 Xiangtai Road, Liuyang economic and Technological Development Zone, Changsha City, Hunan Province, 410300 Patentee after: Changsha Boyuan Medical Technology Co.,Ltd. Address before: 215163 rooms 101 and 201, building 9, No.8 Jinfeng Road, high tech Zone, Suzhou City, Jiangsu Province Patentee before: SUZHOU EVERMED MEDICAL TECHNOLOGY CO.,LTD. |