CN101413946B - Enzyme-linked immunologic detection method of sibutramine hydrochloride - Google Patents
Enzyme-linked immunologic detection method of sibutramine hydrochloride Download PDFInfo
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- CN101413946B CN101413946B CN2008102348521A CN200810234852A CN101413946B CN 101413946 B CN101413946 B CN 101413946B CN 2008102348521 A CN2008102348521 A CN 2008102348521A CN 200810234852 A CN200810234852 A CN 200810234852A CN 101413946 B CN101413946 B CN 101413946B
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- sibutramine hydrochloride
- enzyme
- sibutramine
- dinor
- hydrochloride
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Abstract
An enzyme-linked immunoassay method of sibutramine hydrochloride belongs to the technical field of immunodetection. The enzyme-linked immunosorbent assay method utilizes the immunization of a synthetic bisdesmethyl sibutramine hydrochloride immunogen to obtain a polyclonal antibody, takes the sibutramine hydrochloride as a standard product, takes a conjugate of a hapten of bisdesmethyl sibutramine hydrochloride and OVA as a coating antigen and establishes the indirect competitive enzyme-linked immunosorbent assay method of the sibutramine hydrochloride. The enzyme-linked immunosorbent assay method establishes the indirect competitive ELISA method of the sibutramine hydrochloride and provides a rapid and high-efficient detection method for detecting the residual sibutramine hydrochloride, as the method adopts the polyclonal antibody, the cost is lower, the stability and the repeatability are good, the sensitivity is 0.1ng/ml, the linear range is 0-100ng/ml, while the high specificity and the affinity of the immune reaction lead the ELISA to have very high selectivity and sensitivity.
Description
Technical field
The present invention relates to a kind of slimming drugs method for detecting residue, say so more specifically, belong to technical field of immunoassay a kind of enzyme-linked immune detection method of sibutramine hydrochloride.
Background technology
Sibutramine hydrochloride, English name: Sibutramine Hydrochloride, its chemical name is: (±) N-{1-[1-(4-chlorphenyl) cyclobutyl]-3-methyl butyl }-N, the TMSDMA N dimethylamine hydrochloride monohydrate, CAS 106650-56-0, molecular formula is C
17H
26ClNHClH
2O is the hydrochloride hydrate of sibutramine.These article are by the development of German Kno ll company, and this medicine obtains drugs approved by FDA in November, 1997 and goes on the market in the U.S.; In May, 2000, this medicine obtains National Drug Administration (SDA) approval in China's listing, is used for treatment of obesity.Sibutramine hydrochloride is the slimming drugs of oral central action; It is oral to be absorbed rapidly; Through liver first-pass effect, be two kinds of activated products by the Cytochrome P450 3A4 isodynamic enzyme metabolism of liver, promptly single demethylated sibutramine (secondary amine class; M1) and the dinor-sibutramine (the primary amine class, M2) and the generation effect.The main mechanism of sibutramine interior metabolism product is to suppress the reuptake of norepinephrine, serotonin and dopamine; Increase the concentration of synaptic cleft norepinephrine, serotonin and dopamine; Cause that the appestat feeling of repletion strengthens, quantity of heat production increases; And the release of norepinephrine, serotonin and dopamine is not had obvious influence.Research shows that also sibutramine and amine active metabolite thereof do not have obvious cholinolytic, antihistamine and monoamine oxidase inhibiting effect.Be applicable to motion and the still immitigable obesity of diet control.The common digestive system bad reaction of sibutramine mainly contains dry, apocleisis and constipation, and also having rare digestive system bad reaction is diarrhoea, gastroenteritis and parageusia, and incidence is general, and < 1%, idol has the report of liver and renal damage.Sibutramine has rising blood pressure and the effect of accelerating heart rate, and its nervous system bad reaction has insomnia, headache, giddy, dizziness etc., and unusual, the limb spasm of also can feeling, tension force increase, thinking is unusual, epileptic attack.After in Dec, 2002, " medicine is strong " authentication code is cancelled, and from then on health products are split into medicine and food." Qu Mei ", " Xenical " /> etc. are that master's slimming health product becomes the medicine that receives strict supervision with the chemicals composition, are that the slimming health product of principal ingredient then becomes health food with food.At present, China's slimming health food faces two subject matters, and the one, false, exaggeration of propaganda, the 2nd, add forbidden drug.Exaggerative, the false propaganda problem of health slimming food on the fat-reducing market, national departments concerned has given great attention, and has strengthened law enforcement dynamics, and a collection of have problem exaggerative, false propaganda to be concentrated exposure.Yet with respect to exaggeration of propaganda; In food, add some medicines and structure of modification thing thereof; This type of violated interpolation situation is hidden, and it has not only violated food hygiene law, and owing to the trust of consumer to food, health products security; Without stint is taken this based food, health products, and is healthy very harmful to it.For the supervision of this violated interpolation behavior be controlled at the technical comparison difficulty that just seems.Because of most of supervision and inspection department lacks the corresponding method of inspection and standard items, be difficult to check that this composition in the product is proved conclusively, make product add the forbidden drug situation and be difficult to investigate and prosecute.The main analytical approach of sibutramine hydrochloride has now: high performance liquid chromatography, capillary electrophoresis, LC-MS, LC-MSMS, GC/MS, derivatization fluorescence spectrophotometry, ultraviolet spectroscopy and thin-layered chromatography etc.Though existing instrument detecting method has advantages such as highly sensitive, applied widely, sample pre-treatments is complicated, and required instrument is expensive; Cost is high, and detection time is long, therefore is necessary to study high specificity; The highly sensitive while is cheap, immunoassay technology that can be fast and convenient.This just needs the synthetic immunogene that can produce group-specific antibody.So far; Still not to the report of the how residual immunologic detection method of sibutramine hydrochloride structure, design the metabolin dinor-sibutramine hydrochloride M2 that has synthesized with sibutramine hydrochloride is the haptenic comlete antigen that is used for the sibutramine hydrochloride structure detection for this reason both at home and abroad.
Summary of the invention
The object of the present invention is to provide the residual immunological detection method of a kind of fast detecting sibutramine hydrochloride, and higher sensitivity and specificity are arranged.
Technical scheme of the present invention is following: utilize (a kind of immunogen synthesis method that is applicable to sibutramine hydrochloride) synthetic immunogen immune such as the petty official is transmitted, Li Zhuokun, Ma Wei to obtain polyclonal antibody; As envelope antigen, set up the residual indirect competitive enzyme-linked immunosorbent method of sibutramine hydrochloride with the conjugate of dinor-sibutramine hydrochloride haptens and OVA.
The present invention realizes through following steps:
A kind of enzyme-linked immune detection method of sibutramine hydrochloride; It is characterized in that utilizing synthetic dinor-sibutramine hydrochloride immunogen immune to obtain polyclonal antibody; With the sibutramine hydrochloride is standard items; As envelope antigen, set up the indirect competitive enzyme-linked immunosorbent detection method of sibutramine hydrochloride with the conjugate of dinor-sibutramine hydrochloride haptens and OVA; Step is following:
(1) with carbonate buffer solution dilution envelope antigen 1:5000～1:8000 of 0.05M, add in the ELISA Plate, every hole 100 μ L, 4 ℃ of incubated overnight use the above-mentioned carbonate buffer solution that contains 0.1% gelatin as confining liquid, sealing 2h then;
(2) with PBS the sibutramine hydrochloride standard items are diluted to 0.01,0.1,0.5,1,5,10,20ng/mL series concentration, and the sample of handling well join respectively in the enzyme mark hole separately, add 50 μ L/ holes; With antibody diluent polyclonal antibody is diluted with 1:6000～1:9000 ratio, every then hole adds the good polyclonal antibody of 50 μ L dilution, in 37 ℃ of incubation 1h, then with PBST cleansing solution washing 3～5 times;
(3) the goat-anti rabbit GAR-HRP with horseradish peroxidase-labeled dilutes with 1:3000～1:5000 with antibody diluent, and every then hole adds 100 μ L, washs 3～5 times with the PBST cleansing solution behind 37 ℃ of effect 1h;
(4) every hole adds colour developing liquid 100 μ L, 37 ℃ of colour developing 15min; Add stop buffer 2M sulfuric acid solution, every hole 100 μ L; ELIASA 450nm surveys light absorption value A
450, contrast the sibutramine hydrochloride content of calculating testing sample with the work typical curve.
Configuration colour developing liquid
A liquid: 0.933g citric acid, 3.68g Na
2HPO
412H
2O, 18 μ L30%H
2O
2Be settled to 100mL with ultrapure water;
B liquid: 60mg3,3 ', 5,5 '-tetramethyl benzidine (TMB) is dissolved in the 100mL monoethylene glycol;
Before using A liquid is mixed with the 5:1 volume ratio with B liquid.
More detailed step is:
Main solution preparation
1) preparation phosphate 0.01M (PBS) damping fluid:
Na
2HPO
4·12H
2O 3.62g
KH
2PO
4 0.2g
NaCl 0.2g
KCl 8.0g
Add ultrapure water and be diluted to 1000mL;
2) preparation carbonate (CBS) buffer solution (0.05M) pH9.6
Na
2CO
3 1.59g
NaHCO
3 2.93g
Add ultrapure water and be diluted to 1000mL;
3) preparation PBST solution: the PBS solution that contains 0.05%Tween-20.
4) preparation confining liquid: the carbonate buffer solution that contains 0.1% gelatin.
5) preparation antibody diluent: the PBST solution that contains 0.1% gelatin.
6) colour developing liquid:
A liquid: 0.933g citric acid, 3.68g Na
2HPO
412H
2O, 18 μ L 30%H
2O
2Be settled to 100mL with ultrapure water;
B liquid: 60mg3,3 ', 5,5 '-tetramethyl benzidine (TMB) is dissolved in the 100mL monoethylene glycol;
Before using A liquid is mixed with the 5:1 volume ratio with B liquid.
7) H of stop buffer: 2M
2SO
4
The step of indirect competitive ELISA experimental technique is following:
The methanol solution that in advance standard items of sibutramine hydrochloride is mixed with 100 μ g/mL is as the work mother liquor, and is for use 4 ℃ of preservations.(0.15mol/L NaCl 0.5%Tween-20), prepares serial reaction liquid to preparation PBST solution based on this for 0.01mol/L, pH7.4, in order to dilution competition thing titer and polyclonal antibody.
A, encapsulate: with the coating antigen coated elisa plate of setting concentration, 100 μ L/ holes, 4 ℃ are spent the night.
B, washing: with PBST detersive enzyme target 3 times, each 3min, 200 μ L/ holes dry ELISA Plate then.
C, sealing: contain the damping fluid that encapsulates of 0.1% gelatin, 200 μ L/ holes, 37 ℃ of sealing 2h.
D, washing: same b.
E, competition: with PBST sibutramine hydrochloride is diluted to 0.01,0.1,0.5,1,5,10, the 20ng/mL series concentration, other establishes a PBST blank, 50 μ L/ holes.Every then hole adds the polyclonal antibody of 8100 times of 50 μ L dilutions, in 37 ℃ of incubation 1h.
F, washing: same b.
G, add ELIAS secondary antibody (goat-anti rabbit GAR-HRP, 1:4000), 100 μ L/ holes, 37C reacts 1h.
H, washing: same b.
I, colour developing: add colour developing liquid 100 μ L/ holes, colour developing 15min.
J, termination: add stop buffer 100 μ L/ holes.
K, mensuration: the light absorption value that detects 450nm with ELIASA.
Beneficial effect of the present invention: the present invention has set up the indirect ELISA method of sibutramine hydrochloride; For the sibutramine hydrochloride residue detection provides a kind of detection means rapidly and efficiently; Because what adopt is polyclonal antibody, expense is lower and stable and repeated better.Sensitivity is 0.1ng/mL, and the range of linearity is 0-100ng/mL, half amount of suppression (IC
50) be 9.6ng/mL.Immunoreactive high specific and high-affinity make ELISA have high selectivity and sensitivity, and sample pre-treatment process is simple.
Description of drawings
The standard of Fig. 1 sibutramine hydrochloride suppresses curve.
Specific embodiments
Below further specify the present invention through embodiment.
One, instrument:
TGL-40B table-type low-speed hydro-extractor, Anting Scientific Instrument Factory, Shanghai;
The KFLOW water purification machine, Kai Folong company;
The horizontal shaking table of ZD-9556, granary science and education equipment factory;
Costar96 hole 8 * 12 removable ELISA Plates, the lucky safe bio tech ltd in Shanghai;
MuLtiska Mks ELIASA, Thermo Labsystems company;
Can debug pipettor, Thermo Labsystems company;
Turbine mixer, Shanghai Hu Xi instrumental analysis factory.
Two, reagent:
The goat anti-rabbit igg of horseradish peroxidase-labeled (GAR-HRP), health becomes bio-engineering corporation;
Tetramethyl benzidine (TMB), Huamei Bio-Engrg Co.;
Other reagent are AR.
Three, step
1. immunogene and coating antigen is synthetic
With the mixed anhydride method coupling behind the derivatization, concrete steps are following for synthetic immunogen (dinor-sibutramine hydrochloride haptens and bovine serum albumin(BSA) BSA conjugate) and coating antigen (dinor-sibutramine hydrochloride haptens and ovalbumin OVA conjugate):
1. preparing A liquid: 20mg (0.06627mmol) dinor-sibutramine hydrochloride solution is dissolved in the pyridine solution that 1.5mL is dissolved with 8mg (0.08mmol) succinic anhydride; The lucifuge stirring dried up with nitrogen after 23 hours under the normal temperature then; Add N again; Dinethylformamide DMF: 1, after 1: 1 mixed solvent 1.2mL of 4-dioxane volume ratio fully dissolves, add 21.2 μ L (0.08mmol) tri-n-butylamines and in frozen water, stirred 10 minutes; Add 12 μ L (0.08mmol) isobutyl chlorocarbonate stirring at room reaction 1h again, this liquid is A liquid.
2. prepare B liquid: take by weighing boric acid that 100mg (0.00151mmol) bovine serum albumin(BSA) BSA (or 67.95mg ovalbumin OVA) is dissolved in 5.8mL pH8.5 and receive in the buffer solution cryopreservation.This liquid is B liquid.
3. under stirring at low speed, A liquid dropwise is added drop-wise in the B liquid, stirring reaction 24h under the room temperature, the centrifugal deposition of removing is got supernatant and is promptly obtained the artificial antigen mixed liquor.
4. the artificial antigen mixed liquor is moved in the bag filter, dialysed 4-6 days with the deionized water of 6 * 1L.Use desivac that the liquid in the bag filter is processed powder at last, promptly obtain artificial antigen: dinor-sibutramine hydrochloride-bovine serum albumin (or coating antigen dinor-sibutramine hydrochloride-ovalbumin).
2, ELISA course of reaction:
The antibody titer determination step:
1) coating antigen is encapsulated 96 hole ELISA Plates with carbonate buffer solution as serial dilution, 100 μ L/ holes are in 4 ℃ of refrigerator overnight.Take out ELISA Plate and be back to room temperature next day, and 200 μ LPBST solution are injected in every hole, and the 3min that vibrates on the shaking table firmly gets rid of cleansing solution, does at the thieving paper arsis, continues washing 2 times.Following washing methods is identical.
2) after the abundant washing, with sealing damping fluid sealase target, 200 μ L/ holes, taking-up is dried for use behind the incubation 2h in 37 ℃ of incubation casees.
3) positive serum serial dilution correspondence is joined preceding 7 ranks of ELISA Plate, eighth row adds negative serum, and 100 μ L/ holes are hatched washing behind the 1h, clapped and do for 37 ℃.
4) every hole adds 100 μ L, and the goat-anti rabbit GAR-HRP of the HRP mark of 1:4000 dilution is hatched washing behind the 1h, clapped and do for 37 ℃.
5) every hole adds 100 μ L colour developing liquid (TMB and substrate solution ratio are 1:5), and the 37 ℃ of reactions in dark place 15min takes out every hole, back and adds 100 μ L stop buffers (sulfuric acid of 2mol/L), measures light absorption value A with ELIASA
450
The antibody specificity determination step:
A, encapsulate: with the coating antigen coated elisa plate of setting concentration, 100 μ L/ holes, 4 ℃ are spent the night.
B, washing: with PBST detersive enzyme target three times, each 3min, 200 μ L/ holes dry ELISA Plate then.
C, sealing: contain the carbonate buffer solution of 0.1% gelatin, 200 μ L/ hole .37 ℃ of sealing 2h.
D, washing: same b.
E, competition: with PBST the sibutramine hydrochloride mother liquor is diluted to 0.01,0.05,0.1,1,5,10, the 50ng/mL series concentration, other establishes a PBST blank, 50 μ L/ holes.Every then hole adds the polyclonal antibody of 4000 times of 50 μ L dilutions, in 37 ℃ of incubation 1h.
F, washing: same b.
G, add ELIAS secondary antibody (goat-anti rabbit GAR-HRP, 1:4000), 100 μ L/ holes, 37C reacts 1h.
H, washing: same b.
I, colour developing: add colour developing liquid 100 μ L/ holes, colour developing 15min.
J, termination: add stop buffer 100 μ L/ holes.
K, mensuration: the light absorption value that detects 450nm with ELIASA.
Test findings is following:
1, typical curve: the range of linearity of the antibody test that this experiment obtained is to be 0～100ng/mL.
2, sensitivity: sensitivity is the concentration of the pairing standard items of gained 90% maximum light absorption value, i.e. IC
90Be 0.1ng/mL.
Claims (1)
1. the enzyme-linked immune detection method of a sibutramine hydrochloride; It is characterized in that utilizing synthetic dinor-sibutramine hydrochloride immunogen immune to obtain polyclonal antibody; With the sibutramine hydrochloride is standard items; As envelope antigen, set up the indirect competitive enzyme-linked immunosorbent detection method of sibutramine hydrochloride with the conjugate of dinor-sibutramine hydrochloride haptens and OVA; Step is following:
(1) carbonate buffer solution with 0.05M diluted envelope antigen 1: 5000～1: 8000, add in the ELISA Plate, and every hole 100 μ L, 4 ℃ of incubated overnight use the above-mentioned carbonate buffer solution that contains 0.1% gelatin as confining liquid, sealing 2h then;
(2) with PBS the sibutramine hydrochloride standard items are diluted to 0.01,0.1,0.5,1,5,10, the 20ng/mL series concentration with standard items after the above-mentioned dilution and the sample handled well, joins respectively in the enzyme mark hole separately, adds 50 μ L/ holes; With antibody diluent polyclonal antibody is diluted with 1: 6000～1: 9000 ratio, every then hole adds the good polyclonal antibody of 50 μ L dilution, in 37 ℃ of incubation 1h, then with PBST cleansing solution washing 3～5 times;
(3) the goat-anti rabbit GAR-HRP with horseradish peroxidase-labeled dilutes with 1: 3000～1: 5000 with antibody diluent, and every then hole adds 100 μ L, washs 3～5 times with the PBST cleansing solution behind 37 ℃ of effect 1h;
(4) every hole adds colour developing liquid 100 μ L, 37 ℃ of colour developing 15min; Add stop buffer 2M sulfuric acid solution, every hole 100 μ L; ELIASA 450nm surveys light absorption value A450, with the sibutramine hydrochloride content of calculating testing sample according to the typical curve contrast of standard items preparation;
Said synthetic dinor-sibutramine hydrochloride immunogene is the conjugate with dinor-sibutramine hydrochloride haptens and bovine serum albumin(BSA) BSA coupling acquisition.
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102128923B (en) * | 2010-12-24 | 2013-05-15 | 江南大学 | One-step ELISA (Enzyme Linked Immunosorbent Assay) method for neomycin (NEO) residues in milk |
| CN104076037B (en) * | 2013-03-28 | 2016-07-27 | 北京市药品检验所 | The method of sibutramine in detection goods |
| CN104698176A (en) * | 2015-03-18 | 2015-06-10 | 天津农学院 | Immunoaffinity stir bar for adsorbing chloramphenicol, and preparation method and application thereof |
| CN108693350A (en) * | 2018-03-06 | 2018-10-23 | 山西省食品药品检验所(山西省药品包装材料监测中心) | A kind of sibutramine colloidal gold quick detection device and application thereof |
| CN109022367A (en) * | 2018-08-16 | 2018-12-18 | 江南大学 | The anti-sibutramine monoclonal antibody specific hybridoma cell strain of one plant of secretion and its application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1557802A (en) * | 2004-01-13 | 2004-12-29 | 苏州市玮琪生物科技有限公司 | Sibutramine aromatic salt of organic acid and its preparation process |
| CN1668331A (en) * | 2002-07-18 | 2005-09-14 | 赛托斯生物技术公司 | Hapten-carrier conjugates and uses thereof |
-
2008
- 2008-10-31 CN CN2008102348521A patent/CN101413946B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1668331A (en) * | 2002-07-18 | 2005-09-14 | 赛托斯生物技术公司 | Hapten-carrier conjugates and uses thereof |
| CN1557802A (en) * | 2004-01-13 | 2004-12-29 | 苏州市玮琪生物科技有限公司 | Sibutramine aromatic salt of organic acid and its preparation process |
Non-Patent Citations (3)
| Title |
|---|
| 权伍英.LC-MS/MS检测减肥保健品中的盐酸芬氟拉明和盐酸西布曲明.《中国卫生检验杂志》.2007,第17卷(第1期),10-12. * |
| 王金生.HPLC-PDA法检验保健食品中的盐酸西布曲明.《中国卫生检验杂志》.2007,第17卷(第3期),462,507. * |
| 秦宗会.盐酸西布曲明衍生光度法分析研究.《药物分析杂志》.2006,第26卷(第11期),1656-1658. * |
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