CN1139810C - Method and test paper of smei-quantitative determination of morphine by monoclone antibody immune chromatography - Google Patents
Method and test paper of smei-quantitative determination of morphine by monoclone antibody immune chromatography Download PDFInfo
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- CN1139810C CN1139810C CNB98112724XA CN98112724A CN1139810C CN 1139810 C CN1139810 C CN 1139810C CN B98112724X A CNB98112724X A CN B98112724XA CN 98112724 A CN98112724 A CN 98112724A CN 1139810 C CN1139810 C CN 1139810C
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- morphine
- immunochromatography
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- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 title claims abstract description 85
- 229960005181 morphine Drugs 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 14
- 238000012360 testing method Methods 0.000 title claims abstract description 12
- 238000004587 chromatography analysis Methods 0.000 title abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims abstract description 5
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 5
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 5
- 238000001179 sorption measurement Methods 0.000 claims abstract description 3
- 238000003317 immunochromatography Methods 0.000 claims description 18
- 239000002131 composite material Substances 0.000 claims description 15
- 239000012528 membrane Substances 0.000 claims description 15
- 238000001514 detection method Methods 0.000 claims description 10
- 238000011002 quantification Methods 0.000 claims description 6
- 230000000890 antigenic effect Effects 0.000 claims description 3
- 230000002860 competitive effect Effects 0.000 claims description 2
- 239000011148 porous material Substances 0.000 abstract description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 abstract 2
- 210000002700 urine Anatomy 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 239000002585 base Substances 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
- 238000004458 analytical method Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- ZPUCINDJVBIVPJ-LJISPDSOSA-N cocaine Chemical compound O([C@H]1C[C@@H]2CC[C@@H](N2C)[C@H]1C(=O)OC)C(=O)C1=CC=CC=C1 ZPUCINDJVBIVPJ-LJISPDSOSA-N 0.000 description 2
- OROGSEYTTFOCAN-DNJOTXNNSA-N codeine Chemical compound C([C@H]1[C@H](N(CC[C@@]112)C)C3)=C[C@H](O)[C@@H]1OC1=C2C3=CC=C1OC OROGSEYTTFOCAN-DNJOTXNNSA-N 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- -1 stable Chemical compound 0.000 description 2
- 230000009182 swimming Effects 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 1
- ZQBMUHABRSEAIK-UXBLZVDNSA-N 1-[4-[(e)-3-phenylprop-2-enyl]piperazin-1-yl]butan-1-one Chemical compound C1CN(C(=O)CCC)CCN1C\C=C\C1=CC=CC=C1 ZQBMUHABRSEAIK-UXBLZVDNSA-N 0.000 description 1
- USSIQXCVUWKGNF-UHFFFAOYSA-N 6-(dimethylamino)-4,4-diphenylheptan-3-one Chemical compound C=1C=CC=CC=1C(CC(C)N(C)C)(C(=O)CC)C1=CC=CC=C1 USSIQXCVUWKGNF-UHFFFAOYSA-N 0.000 description 1
- AKJDEXBCRLOVTH-UHFFFAOYSA-N Carbetapentane citrate Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O.C=1C=CC=CC=1C1(C(=O)OCCOCCN(CC)CC)CCCC1 AKJDEXBCRLOVTH-UHFFFAOYSA-N 0.000 description 1
- GJSURZIOUXUGAL-UHFFFAOYSA-N Clonidine Chemical compound ClC1=CC=CC(Cl)=C1NC1=NCCN1 GJSURZIOUXUGAL-UHFFFAOYSA-N 0.000 description 1
- GVGLGOZIDCSQPN-PVHGPHFFSA-N Heroin Chemical compound O([C@H]1[C@H](C=C[C@H]23)OC(C)=O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4OC(C)=O GVGLGOZIDCSQPN-PVHGPHFFSA-N 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-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
- XADCESSVHJOZHK-UHFFFAOYSA-N Meperidine Chemical compound C=1C=CC=CC=1C1(C(=O)OCC)CCN(C)CC1 XADCESSVHJOZHK-UHFFFAOYSA-N 0.000 description 1
- JEYCTXHKTXCGPB-UHFFFAOYSA-N Methaqualone Chemical compound CC1=CC=CC=C1N1C(=O)C2=CC=CC=C2N=C1C JEYCTXHKTXCGPB-UHFFFAOYSA-N 0.000 description 1
- 229920000297 Rayon Polymers 0.000 description 1
- 241001104043 Syringa Species 0.000 description 1
- 235000004338 Syringa vulgaris Nutrition 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- VREFGVBLTWBCJP-UHFFFAOYSA-N alprazolam Chemical compound C12=CC(Cl)=CC=C2N2C(C)=NN=C2CN=C1C1=CC=CC=C1 VREFGVBLTWBCJP-UHFFFAOYSA-N 0.000 description 1
- 229960004538 alprazolam Drugs 0.000 description 1
- 229940025084 amphetamine Drugs 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 229960001736 buprenorphine Drugs 0.000 description 1
- RMRJXGBAOAMLHD-IHFGGWKQSA-N buprenorphine Chemical compound C([C@]12[C@H]3OC=4C(O)=CC=C(C2=4)C[C@@H]2[C@]11CC[C@]3([C@H](C1)[C@](C)(O)C(C)(C)C)OC)CN2CC1CC1 RMRJXGBAOAMLHD-IHFGGWKQSA-N 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 229940098391 carbetapentane citrate Drugs 0.000 description 1
- 239000003795 chemical substances by application Substances 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
- 229960002896 clonidine Drugs 0.000 description 1
- QZUDBNBUXVUHMW-UHFFFAOYSA-N clozapine Chemical compound C1CN(C)CCN1C1=NC2=CC(Cl)=CC=C2NC2=CC=CC=C12 QZUDBNBUXVUHMW-UHFFFAOYSA-N 0.000 description 1
- 229960004170 clozapine Drugs 0.000 description 1
- 229960003920 cocaine Drugs 0.000 description 1
- 229960004126 codeine Drugs 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 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
- 230000006378 damage Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 229960002069 diamorphine Drugs 0.000 description 1
- BRTSNYPDACNMIP-FAWZKKEFSA-N dihydroetorphine Chemical compound O([C@H]1[C@@]2(OC)CC[C@@]34C[C@@H]2[C@](C)(O)CCC)C2=C5[C@]41CCN(C)[C@@H]3CC5=CC=C2O BRTSNYPDACNMIP-FAWZKKEFSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- HYPPXZBJBPSRLK-UHFFFAOYSA-N diphenoxylate Chemical compound C1CC(C(=O)OCC)(C=2C=CC=CC=2)CCN1CCC(C#N)(C=1C=CC=CC=1)C1=CC=CC=C1 HYPPXZBJBPSRLK-UHFFFAOYSA-N 0.000 description 1
- 229960004192 diphenoxylate Drugs 0.000 description 1
- 206010013663 drug dependence Diseases 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229960002179 ephedrine Drugs 0.000 description 1
- 229960002428 fentanyl Drugs 0.000 description 1
- PJMPHNIQZUBGLI-UHFFFAOYSA-N fentanyl Chemical compound C=1C=CC=CC=1N(C(=O)CC)C(CC1)CCN1CCC1=CC=CC=C1 PJMPHNIQZUBGLI-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 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
- 238000003018 immunoassay Methods 0.000 description 1
- 229960005209 lofexidine Drugs 0.000 description 1
- KSMAGQUYOIHWFS-UHFFFAOYSA-N lofexidine Chemical compound N=1CCNC=1C(C)OC1=C(Cl)C=CC=C1Cl KSMAGQUYOIHWFS-UHFFFAOYSA-N 0.000 description 1
- 229960001797 methadone Drugs 0.000 description 1
- 229960002803 methaqualone 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
- 210000000056 organ Anatomy 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 229960000482 pethidine Drugs 0.000 description 1
- WRLGYAWRGXKSKG-UHFFFAOYSA-M phenobarbital sodium Chemical compound [Na+].C=1C=CC=CC=1C1(CC)C(=O)NC([O-])=NC1=O WRLGYAWRGXKSKG-UHFFFAOYSA-M 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000011117 substance-related disease Diseases 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- QDWJJTJNXAKQKD-UHFFFAOYSA-N trihexyphenidyl hydrochloride Chemical compound Cl.C1CCCCC1C(C=1C=CC=CC=1)(O)CCN1CCCCC1 QDWJJTJNXAKQKD-UHFFFAOYSA-N 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
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- Peptides Or Proteins (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to a method and test paper for detecting morphine semi-quantitatively in a one-step immune chromatography method, which has the technical scheme that morphine in liquid to be detected and morphine marked on the surface of colloidal gold are competitively combined with morphine monoclonal antibodies accurately and quantitatively dropped on a chromatography film in a horizontal strip form; the content of the morphine in the liquid to be detected is determined by the number of stripes displayed on the chromatography film; the more the stripes are, the higher the content of the morphine in the liquid to be detected is; the bottom plate of the test paper and an immune chromatography complex film are sequentially superposed; colloidal gold is positioned at one end of the bottom plate and is in contact with the complex film; the chromatography film has good adsorption force to protein and has good wettability; the chromatography film is made of high molecular materials having a certain micro pore diameter.
Description
Technical field: the present invention discloses a kind of method and test paper of smei-quantitative determination of morphine by monoclone antibody immune chromatography.
Background technology: the detection method to morphine in drug addict's body fluid can be divided three major types, i.e. chemical colour reaction analysis (as AC method etc.), stratographic analysis (as thin-layer chromatography, gas-matter coupling etc.) and immunoassay (as radioimmunology, immunochromatographic method etc.).The immunochromatography single stage method is to be set up by American scholar the early 1990s, and this method need not handled with its sample, and easy to be quick, characteristics such as suitable on-the-spot detection bringing into play very important effect in the drugs testing, but this method can only be qualitative, can not be quantitative; And its quality control system is another set of antigen-antibody, can not reflect the response situation of detection system truly; In addition, its reaction pattern shows as the positive and does not develop the color at detection zone, and negative for developing the color, this kind reaction pattern does not meet the thinking set of common result of determination.
Summary of the invention: the object of the invention is to overcome above-mentioned technical deficiency and a kind of high specificity is provided, and is highly sensitive, simple in structure, easy for operation, tentatively the method and the test paper of the smei-quantitative determination of morphine by monoclone antibody immune chromatography of detection by quantitative.
Fundamental purpose of the present invention is achieved in that by morphine in the liquid to be checked and the morphine that is marked at the collaurum surface and at least two accurate quantifications and is the horizontal stripe shape and selects the morphine monoclonal antibody band that schedules on the immunochromatography composite membrane competitive the combination taken place, by being shown in the striped quantity on the immunochromatography composite membrane, to determine the content of morphine in the liquid to be checked, fringe number is many more, show that morphine content is high more in the liquid to be checked, thereby finish the half-quantitative detection of morphine.
Another object of the present invention is achieved in that test paper comprises immunochromatography composite membrane, antigenic mark collaurum, base plate, thieving paper, base plate, immunochromatography composite membrane are stacked successively, one end of collaurum place base plate contacts with the immunochromatography composite membrane, the immunochromatography composite membrane is selected the morphine monoclonal antibody band that fixed at least two accurate quantifications are the horizontal stripe shape by protein being had good adsorption power and infiltrating micropore diameter macromolecular material is made on the immunochromatography composite membrane.
Embodiment: embodiments of the invention are summarized as follows:
One, the preparation of mark collaurum: get 1% gold chloride 1ml and be diluted to 100ml, add a certain amount of reductive agent under the state boiling, make collaurum with DDW.Be with morphine-6-O-half succinyl-bovine serum albumin(BSA) (Mor-BSA) collaurum to be carried out mark under the condition of 7-8.2 at pH value, carry out purifying with supercentrifugal process again, the accurate quantification freeze drying of instiling on glass fibre is standby.
Two, the special-purpose polymer compound film preparation of immunochromatography
At first make transparent backing film with a kind of macromolecular material, it is strong to protein bound power to cover one deck on backing film, the chromatographic film with certain pore size that wetting property is good.The effect of backing film is to stop organic solvent in the viscose binder to the destruction of protein on the chromatographic film.
Three, the point sample method of chromatographic film
With certain density morphine monoclonal antibody accurate quantification, be horizontal stripe shape point on chromatographic film with a determining deviation, use for detecting.
Four, the key technical indexes
1 specificity: only morphine class material is reacted;
2, sensitivity: 0.3 microgram morphine/milliliter sample;
3, sxemiquantitative scope: 〉=0.3 microgram～＞1.2 micrograms;
4, quick: observations in 10 minutes;
5, easy: anyone by specification can be operated;
6, be not subjected to such environmental effects.
Five, the application of test paper
1, to the detection of negative sample: three urine to be measured are splashed into well, keep flat observations after 5 minutes, have only a colour band negative.
2, the detection of positive sample: three urine to be measured are splashed into well, keep flat observations after 5 minutes, two above colour bands are arranged, and positive (〉=0.3 microgram/ml), three colour bands are about 0.8 microgram/ml, and four colour bands are 〉=1.2 micrograms/ml.
3, cross reaction: test paper and heroin and codeine have cross reaction, and intensity is similar to morphine; With methadone, naloxone, buprenorphine, meperidine, dihydroetorphine, fentanyl, diphenoxylate, cocaine, amphetamine, eastern Liang if alkali, ephedrine, haloperole, chlorpromazine, bucinnazine, fenazil, sodium phenobarbital, stable, lofexidine, methaqualone, somedon, clonidine, artane, caffeine, carbetapentane citrate, alprazolam, the equal no cross reaction of Clozapine.
Six, test paper reaction principle
Freeze and on collaurum, be marked with morphine antigen, be fixed with the morphine monoclonal antibody on the polymer compound film.When in the tested urine during no morphine, urine dilution antigenic mark collaurum also promotes its swimming on composite membrane, and when running into article one morphine monoclonal antibody, the morphine antigen on the collaurum will react fully with it, forms a macroscopic lilac band.When having morphine to exist in the tested urine, morphine in the urine will combine with article one morphine monoclonal antibody competitively with the morphine on the collaurum, make the collaurum of the anti-note of morphine not tackled by article one monoclonal antibody fully, can only continue swimming forward, show two colour bands with the reaction of second monoclonal antibody band.In like manner, morphine content is many in the urine, and colour developing tape number is also just many more, thereby can carry out half-quantitative detection to morphine.
In sum, high specificity of the present invention, highly sensitive, simple in structure, easy for operation, can effectively detect the roughly content of drugs in the liquid to be detected, be adapted to the instrument that procuratorial, judicial and public security organ, drug addiction treatment mechanism, hospital, employing unit, drug addict family members etc. detect as drugs.
Claims (2)
1, a kind of method of immunochromatography smei-quantitative determination of morphine, the morphine that it is characterized in that morphine in the liquid to be checked and be marked at the collaurum surface and at least two accurate quantifications also are the horizontal stripe shape and select the morphine monoclonal antibody band that schedules on the immunochromatography composite membrane competitive the combination taken place, by being shown in the striped quantity on the immunochromatography composite membrane, to determine the content of morphine in the liquid to be checked, fringe number is many more, show that morphine content is high more in the liquid to be checked, thereby finish the half-quantitative detection of morphine.
2, a kind of test paper of immunochromatography smei-quantitative determination of morphine, comprise immunochromatography composite membrane, antigenic mark collaurum, base plate, thieving paper, base plate, immunochromatography composite membrane are stacked successively, one end of collaurum place base plate contacts with the immunochromatography composite membrane, protein is had good adsorption power to the immunochromatography composite membrane and infiltrating micropore diameter macromolecular material is made, it is characterized in that, on the immunochromatography composite membrane, select the morphine monoclonal antibody band that fixed at least two accurate quantifications are the horizontal stripe shape.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB98112724XA CN1139810C (en) | 1998-11-10 | 1998-11-10 | Method and test paper of smei-quantitative determination of morphine by monoclone antibody immune chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB98112724XA CN1139810C (en) | 1998-11-10 | 1998-11-10 | Method and test paper of smei-quantitative determination of morphine by monoclone antibody immune chromatography |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1259668A CN1259668A (en) | 2000-07-12 |
| CN1139810C true CN1139810C (en) | 2004-02-25 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB98112724XA Expired - Fee Related CN1139810C (en) | 1998-11-10 | 1998-11-10 | Method and test paper of smei-quantitative determination of morphine by monoclone antibody immune chromatography |
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| Country | Link |
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| CN (1) | CN1139810C (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101526532A (en) * | 2008-06-10 | 2009-09-09 | 陈桂勇 | Narcotic saliva detection |
| CN101551334B (en) * | 2009-03-05 | 2011-02-09 | 公安部物证鉴定中心 | Drug morphia molecular recognition sensitivity chip of optical sensor and producing method thereof |
| CN102393464B (en) * | 2011-10-25 | 2014-01-15 | 广东省药品检验所 | Preparation method of test strip for morphine detection and application thereof |
| CN102590178A (en) * | 2012-03-19 | 2012-07-18 | 中国政法大学 | Method for detecting drug morphine through surface enhanced Raman spectroscopy |
| CN109813886A (en) * | 2017-11-20 | 2019-05-28 | 丹阳亿太生物科技发展有限公司 | A kind of Rapid detection test strip detecting chlorpromazine medicament residue |
-
1998
- 1998-11-10 CN CNB98112724XA patent/CN1139810C/en not_active Expired - Fee Related
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| Publication number | Publication date |
|---|---|
| CN1259668A (en) | 2000-07-12 |
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