A paroxetine hapten and artificial antigen and preparation method and application thereof

Abstract

The present invention discloses a paroxetine hapten and an artificial antigen and a preparation method and application thereof. The paroxetine hapten of chemical formula (I) and the paroxetine artificial antigen of chemical formula (II) are synthesized using 2-hydroxyparoxetine as a precursor. The obtained antigens have good immunogenicity and reactivity, and the corresponding antibodies are easily obtained when performing animal immunization, and can be used to prepare paroxetine colloidal gold detection products.

Description

Technical Field

[0001] The present invention relates to the field of paroxetine detection, and in particular to a paroxetine hapten and an artificial antigen, and a preparation method and application thereof. Background Art

[0002] Paroxetine is a 5-HT reuptake inhibitor antidepressant (SSRIs) that can selectively inhibit the reuptake of 5-hydroxytryptamine (5-HT), increase the concentration of 5-HT in the synaptic cleft, and thus produce an antidepressant effect. It only weakly inhibits the reuptake of norepinephrine and dopamine. This drug does not act on adrenaline α1, α2 or β receptors, nor does it bind to dopamine D2 or histamine H1 receptors, and does not inhibit monoamine oxidase. This drug can be widely distributed in various tissues and organs, and only 1% appears in the systemic circulation. The protein binding rate is as high as 95%. The drug is metabolized by the liver P450 isoenzyme. Most of paroxetine is excreted in the urine through the kidneys; about 36% is excreted in the feces; it can also be excreted through breast milk. Paroxetine is the strongest reuptake agent in the SSRI class, but it is also the only drug with a mild anticholinergic effect.

[0003] For drug abuse, medical staff can detect paroxetine in urine to help diagnose paroxetine dependence and the rehabilitation of paroxetine. At present, high performance liquid chromatography (HPLC), liquid mass spectrometry (LC-MS) and other methods are commonly used to detect paroxetine-related substances. This method has strong separation and analysis capabilities, high sensitivity, reliable and accurate results, but it has a complex structure, high requirements for environmental temperature and humidity, and high maintenance costs. The mass spectrometer is a high-precision instrument that requires specially trained technicians to operate, and the test speed is slow, so it is not conducive to the widespread promotion of drug abuse detection. Therefore, it is necessary to develop a simple and efficient method for detecting paroxetine substances. Summary of the invention

[0004] The first purpose of the present invention is to overcome the problems of high requirements on instruments and operators, difficulty in promotion and slow detection speed of paroxetine detection by HPLC and LC-MS in the prior art, and to provide a paroxetine artificial hapten that can specifically identify paroxetine.

[0005] A paroxetine hapten, whose molecular structure is shown in formula (I),

[0006]

[0007] There is no active group in the 2-hydroxyparoxetine molecule that can be connected to the carrier protein, so it is modified and connected with a suitable spacer arm to produce a carboxyl group or an amino group to facilitate coupling to the carrier protein.

[0008] The second object of the present invention is to provide a method for preparing a paroxetine artificial hapten, which adopts the following technical scheme:

[0009] Step (1), dissolving 2-hydroxyparoxetine in N,N-dimethylformamide (DMF), adding triethylamine and di-tert-butyl dicarbonate, heating under reflux at 45-50° C., cooling the reaction solution to room temperature after the reaction is complete, and evaporating the solvent under reduced pressure to obtain an oily substance A;

[0010] Step (2), dissolving the oil phase substance A in pyridine, then adding glutaric anhydride and 4-dimethylaminopyridine (DMAP), heating under reflux at 98-100° C., cooling the reaction solution to room temperature after the reaction is complete, and evaporating the solvent under reduced pressure to obtain an oily residue; adding dichloromethane to the oily residue, washing with purified water and saturated NaCl respectively, collecting the dichloromethane phase, drying, filtering, and evaporating under reduced pressure to obtain an oily substance B;

[0011] Step (3), dissolving the oil B in a mixed solution of HCl and 1,4-dioxane, heating with stirring under reflux at 98-100° C., and evaporating the solvent under reduced pressure to obtain an oil C;

[0012] Step (4), dissolving the oily substance C in benzene, placing in an ice-water bath, adding trifluoroacetic anhydride, stirring at room temperature for a period of time, and then heating and stirring at 78-81° C. and reflux. After the reaction is complete, the reaction solution is dried and separated to obtain the oily substance D, i.e., the artificial hapten of paroxetine.

[0013] Preferably, in step (1), the molar ratio of 2-hydroxyparoxetine to triethylamine and di-tert-butyl dicarbonate is 1:2:2;

[0014] Preferably, in step (1), the reaction temperature is 45° C. and the reaction time is 14.5-16 h, more preferably 15 h;

[0015] Preferably, in step (2), the molar ratio of oily substance A, glutaric anhydride and 4-dimethylaminopyridine (DMAP) is (10-12): (4-5.5): (1-2), more preferably 10:5:1;

[0016] Preferably, in step (2), the reaction temperature is 100° C. and the reaction time is 16-19 h, more preferably 18 h;

[0017] Preferably, in step (3), 7.5 nmol of oil B is added to each mL of the mixed solution;

[0018] Preferably, in step (3), the heating temperature is 100° C., and the stirring reflux reaction time is 5-6.5 h, more preferably 6 h;

[0019] Preferably, in step (3), the concentration of HCl in the mixed solution is 2 mol / L;

[0020] Preferably, in step (4), 0.056 mmol of oily substance C is added to each mL of benzene;

[0021] Preferably, in step (4), the stirring time is 0.8-1.2 h, more preferably 1 h;

[0022] Preferably, in step (4), the molar ratio of the oily substance C to trifluoroacetic anhydride is (1-1.3):(1.8-2.4), more preferably 1:2;

[0023] Preferably, in step (4), the heating temperature is 80° C., and the stirring reflux reaction time is 2.6-3.0 h, more preferably 3 h.

[0024] In the synthesis reaction of paroxetine hapten, thin layer chromatography can be used to monitor the formation of brown oil to determine whether the reaction is complete. The volume ratio of each component in the chromatographic solution used is 95% ethanol: dichloromethane: 1,4-dioxane: ammonia water = 8:10:1:1, and the product Rf = 0.4-0.5.

[0025] The third object of the present invention is to provide an artificial paroxetine antigen, which is obtained by coupling the hapten of formula (I) with a carrier protein; it has the molecular structural characteristics of paroxetine, can be recognized by animal immune cells, and produce antibodies that can specifically bind to paroxetine. Its molecular structure is shown in formula (II):

[0026]

[0027] Where R represents carrier protein;

[0028] Preferably, the carrier protein is one of bovine serum albumin, bovine gamma globulin, bovine thyroglobulin, keyhole limpet hemocyanin and chicken ovalbumin.

[0029] The fourth object of the present invention is to provide a method for preparing a paroxetine artificial antigen, using the following technical scheme:

[0030] Step (1), dissolving the paroxetine artificial hapten in N,N-dimethylformamide DMF, adding N-hydroxysuccinimide NHS and cyclohexylcarbodiimide, stirring at room temperature for 15-18 hours, and centrifuging to obtain the supernatant to obtain liquid A;

[0031] Step (2), dissolving the carrier protein in PBS buffer as liquid B, wherein the concentration of the carrier protein in liquid B is 4-9 mg / ml;

[0032] Step (3), adding liquid A dropwise to liquid B under stirring, and standing overnight at low temperature to obtain an artificial antigen mixture;

[0033] Step (4), dialyzing the artificial antigen mixture obtained in step (3) in alkaline dialysate for multiple times, and then transferring it into PBS buffer for multiple dialysates. After the dialysis is completed, the supernatant is centrifuged to obtain the paroxetine artificial antigen.

[0034] Preferably, in step (1), the molar ratio of the paroxetine artificial hapten, N-hydroxysuccinimide, and cyclohexylcarbodiimide is (1.3-1.4):1.9:1.9;

[0035] Preferably, in step (2), the concentration of carrier protein in liquid B is 5 mg / ml;

[0036] Preferably, in step (3), the volume ratio of liquid A to liquid B is 1:5;

[0037] Preferably, in step (3), the low temperature is 4°C;

[0038] Preferably, in step (4), the alkaline dialysate is a sodium carbonate solution with a pH of 11.05-12.05; the PBS buffer has a concentration of 0.1 mol / L and a pH of 7.2-7.4;

[0039] Preferably, in step (4), the dialyzate is used for 2-3 times, and each dialysis time is 24 hours;

[0040] Preferably, in step (4), the dialyzate is performed in PBS buffer for 6-7 times, and each dialysis time is greater than 2 hours.

[0041] Paroxetine hapten reacts with N-hydroxysuccinimide and N,N-cyclohexylcarbodiimide to obtain active ester, which is coupled with carrier protein to obtain paroxetine artificial antigen. This reaction process protects the active site of paroxetine, increases the immunogenicity and reactivity of the antigen, and is easy to obtain corresponding antibodies during animal immunization, providing a guarantee for the preparation of subsequent detection reagents. The supernatant obtained by centrifugation after the dialysis is completed can be scanned by ultraviolet to identify whether the hapten is successfully coupled with BSA.

[0042] The fifth object of the present invention is to provide a paroxetine polyclonal antibody obtained by immunizing animals with the artificial antigen of formula (II).

[0043] The preparation steps of the paroxetine polyclonal antibody include: diluting the paroxetine artificial antigen to 1.67 mg / mL with PBS buffer, emulsifying with Freund's adjuvant at a volume ratio of 1:1, and then performing multiple subcutaneous injections on the back of rabbits, with each injection volume of 500 ug, taking a small amount of blood for titer detection after 7 days to confirm whether the immunization is successful, and collecting serum after the immunization is successful, purifying and preparing the paroxetine polyclonal antibody.

[0044] The sixth object of the present invention is to provide a paroxetine colloidal gold test paper, comprising a test line, a quality control line, and a binding pad, wherein the test line is obtained by using paroxetine artificial antigen as a raw material for spot filming;

[0045] Preferably, the quality control line is obtained by spotting sheep anti-mouse IgG or sheep anti-rabbit IgG as raw material, and the conjugate pad is obtained by spraying paroxetine polyclonal antibody-colloidal gold complex on the conjugate pad.

[0046] The paroxetine colloidal gold test strip of the present invention has high detection sensitivity, and the detectable concentration is up to 500ng / mL. Since the detection principle of the paroxetine detection reagent strip of the present invention uses a competitive method, the color intensity of the detection line is negatively correlated with the concentration of paroxetine in the sample. At the same time, colloidal gold labeling is used to perform qualitative detection. The user observes the aggregation and color development of colloidal gold on the NC membrane with the naked eye. When the quality control line appears, the T line appears, which is negative, and the T line does not appear, which is positive.

[0047] Preferably, the quality control line and the detection line are prepared by the following process: using 1.0 mg / mL of goat anti-rabbit IgG and 0.1 mg / mL of goat anti-mouse IgG, spraying them on a nitrocellulose membrane at a spray volume of 1.0 μL / cm as the quality control line, and using 0.3 mg / mL of paroxetine antigen, spraying them on a nitrocellulose membrane at a spray volume of 1.0 μL / cm as the detection line.

[0048] Preferably, the preparation steps of the paroxetine polyclonal antibody-colloidal gold complex include: adjusting the pH value of the colloidal gold solution to 7.6, adding the paroxetine polyclonal antibody solution, adding 1% PEG2000 buffer after stirring, centrifuging at 100000g and 4°C for 60min, removing the precipitate and resuspending with a buffer containing PEG2000, repeating 2 to 3 times to obtain a colloidal gold labeled protein, then purifying the colloidal gold labeled protein by gel filtration, eluting with a PBS buffer containing BSA and sodium azide, and collecting to obtain the paroxetine polyclonal antibody-colloidal gold complex.

[0049] Preferably, the preparation step of the conjugate pad comprises: diluting the paroxetine polyclonal antibody-colloidal gold complex in a PBS buffer containing bovine serum albumin to 1 / 4 to 1 / 3 of the initial OD value of the complex, and spraying it evenly on the conjugate pad at a spray volume of 1.0 μL / cm.

[0050] Preferably, the preparation of the conjugate pad further comprises adding 50 mg / mL trehalose and 200 mg / mL sucrose to the PBS buffer containing bovine serum albumin.

[0051] The activity of the paroxetine polyclonal antibody-colloidal gold complex can be effectively maintained by adding sucrose and trehalose, the shelf life of the binding pad can be extended, and the product performance can be improved.

[0052] The present invention has the following beneficial effects:

[0053] (1) Paroxetine hapten and paroxetine artificial antigen are synthesized using 2-hydroxyparoxetine as a precursor. The obtained antigen has good immunogenicity and reactivity, and the corresponding antibody is easily obtained when performing animal immunization. The antigen can also be used to prepare paroxetine colloidal gold-test paper;

[0054] (2) The paroxetine colloidal gold-test paper provided by the present invention has high sensitivity, a colloidal gold detection limit of 500 ng / mL, a negative correlation between the color intensity of the test line and the concentration of paroxetine, and can quickly detect paroxetine in urine, blood, and saliva. It can be qualitatively detected by the naked eye and is more convenient and faster than liquid chromatography-tandem mass spectrometry, has low instrument requirements, is easy to operate, and is conducive to the widespread promotion of drug abuse detection. DETAILED DESCRIPTION

[0055] The present invention is further described below in conjunction with specific implementation methods.

[0056] The PBS buffer used in the following examples is prepared by dissolving 14.5 g of disodium hydrogen phosphate dodecahydrate, 43.875 g of sodium chloride, and 1.495 g of sodium dihydrogen phosphate dihydrate in double distilled water to 5.0 L, 0.1 M, pH 7.4; the preparation process of the alkaline dialysate is as follows: a sodium carbonate aqueous solution with a mass fraction of 0.5% is adjusted to pH = 12.00 with a 2 mol / L NaOH solution.

[0057] Example 1

[0058] (1) Preparation of artificial hapten of paroxetine:

[0059] A. Weigh 200 mg (0.58 mmol) of 2-hydroxyparoxetine into a 50 mL single-necked round-bottom flask, add 5 mL of DMF to dissolve, then add 160 μL (1.16 mmol) of triethylamine and 266 μL (1.16 mmol) of di-tert-butyl dicarbonate, place in a 45°C oil bath and stir under reflux for 15 hours. TLC detection shows that the raw materials have been completely reacted; terminate the reaction, cool the reaction solution to room temperature, and pump it dry with an oil pump in a 60°C water bath to obtain a light yellow oil A: 267 mg, which is directly used for the next step of reaction.

[0060] B. Place 267 mg of the pale yellow oil A in the previous step in a 50 mL single-necked round-bottom flask, add 15 mL of pyridine to dissolve, then add 132 mg (1.16 mmol) of glutaric anhydride and 27 mg of DMAP, stir and place in an oil bath at 100°C, stir and reflux for 18 hours, and most of the reaction is complete as detected by TLC; stop the reaction, cool the reaction solution to room temperature, evaporate the solvent under reduced pressure to obtain a brown oily residue, add 30 mL of dichloromethane, wash the dichloromethane phase with 30 mL*2 purified water and 30 mL of saturated brine, respectively, collect the dichloromethane phase, dry it with anhydrous sodium sulfate, filter, and evaporate to dryness under reduced pressure to obtain a brown oily substance. The brown oil was separated by TLC (developing solvent: 95% ethanol: dichloromethane: 1,4-dioxane: ammonia water = 8:10:1:1), and the product spots with Rf = 0.4-0.5 were collected to obtain 168 mg (0.30 mmol) of brown oil B.

[0061] C. Dissolve 168 mg (0.30 mmol) of the brown oil B in 20 mL of 2 mol / L HCl in 1,4-dioxane, place in an oil bath at 100° C. and stir under reflux for 6 hours. After TLC analysis, the reaction is almost complete. Stop the reaction and evaporate the solvent under reduced pressure to obtain 130 mg (0.28 mmol) of brown oil C.

[0062] D. Dissolve 130 mg (0.28 mmol) of brown oil C in 5 mL of benzene, place in an ice-water bath, slowly add 78 μL (0.56 mmol) of trifluoroacetic anhydride, react at room temperature with stirring for 1 hour, transfer to an oil bath at 80°C and stir under reflux for 3 hours, stop the reaction, dry the reaction solution, and separate by TLC to obtain 146 mg (0.26 mmol) of a brown oil, i.e., the artificial hapten of paroxetine.

[0063] H NMR spectrum data:

[0064] 1H-NMR (400MHz, CDCl3): δ1.79-1.95(m,4H), δ2.23-2.38(m,5H), δ2.77(m,1H), δ3.3-3.34(t,4H) , δ3.9(d,2H), δ6.17-6.22(t,2H), δ6.55(d,1H), δ6.89-7.11(m,4H), δ8.37(s,1H), δ10.52(s,1H).

[0065] C NMR spectrum data:

[0066] 13C-NMR (100MHz): δ20.2(CH2), δ29.7(CH2), δ33.0(CH), δ33.3(CH2), δ35.1(CH ), δ35.2(CH2), δ43.6(CH2), δ44.3(CH2), δ71.5(CH2), δ100.8(CH), δ107.5(CH), δ112.7(CH), δ114.6(2CH), δ115.7(CH), δ123.1(C), δ130.3(2CH), δ136.7(C), δ1 39.2(C), δ148.2(C), δ152.1(C), δ158.9(C), δ172.0(C), δ172.7(C), δ177.0(C).

[0067] (2) Preparation of paroxetine artificial antigen:

[0068] A. Take 130 mg of paroxetine artificial hapten (0.28 mmol) and place it in a 25 mL single-necked round-bottom flask, add N,N-dimethylformamide (DMF), then add 43 mg (0.38 mmol) of N-hydroxysuccinimide (NHS) and 78 mg (0.38 mmol) of cyclohexylcarbodiimide (DCC), stir at room temperature for 16 hours, and after the reaction is completed, centrifuge at 8000 r / min and 4°C for 15 min, and take the supernatant as liquid A.

[0069] B. Weigh 0.160 g of bovine serum albumin (BSA) and dissolve it in 32 mL of PBS solution to obtain a 5 mg / mL BSA solution, which is recorded as solution B.

[0070] C. Under rapid stirring, slowly add liquid A to liquid B, the volume ratio of liquid A to liquid B is 1:5, and the resulting mixture is stored at 4°C overnight to obtain an artificial antigen mixture. UV scanning is used to identify whether the hapten and BSA are successfully coupled;

[0071] D. Transfer the successfully coupled artificial antigen mixture into a dialysis bag, dialyze it in alkaline dialysis solution for 24 hours under stirring conditions, and repeat the dialysis twice;

[0072] E. The alkaline dialyzed artificial antigen mixture was dialyzed in PBS solution for 7 times, each dialysis time was 2 hours. After the dialysis, centrifugation was performed and the supernatant was taken to obtain the paroxetine artificial antigen.

[0073]

[0074] Example 2

[0075] (1) Preparation of paroxetine artificial hapten: The molar ratio of 2-hydroxyparoxetine to triethylamine and di-tert-butyl dicarbonate was changed to 1:2:2, and the other preparation conditions were the same as those in Example 1 to prepare the paroxetine artificial hapten;

[0076] (2) Preparation of paroxetine artificial antigen: The molar ratio of paroxetine hapten, N-hydroxysuccinimide and cyclohexylcarbodiimide was changed to 1.3:1.9:1.9, and the other preparation conditions were the same as those in Example 1 to prepare the paroxetine artificial antigen.

[0077] Example 3

[0078] (1) Preparation of paroxetine artificial hapten: The molar ratio of 2-hydroxyparoxetine to triethylamine and di-tert-butyl dicarbonate was changed to 1.2:2:2, and the other preparation conditions were the same as those in Example 1 to prepare the paroxetine artificial hapten;

[0079] (2) Preparation of paroxetine artificial antigen: The molar ratio of paroxetine hapten, N-hydroxysuccinimide and cyclohexylcarbodiimide was changed to 1.4:1.9:1.9, and the other preparation conditions were the same as those in Example 1 to prepare the paroxetine artificial antigen.

[0080] Example 4: Preparation of paroxetine polyclonal antibodies:

[0081] The paroxetine antigen obtained in Example 1 was diluted to 1.67 mg / mL with PBS buffer, emulsified with Freund's adjuvant at a volume ratio of 1:1, and then injected subcutaneously at multiple points on the back of rabbits, with each injection volume of 500 ug. After 7 days, a small amount of blood was taken for titer detection to confirm whether the immunization was successful. After successful immunization, serum was collected and purified to obtain paroxetine polyclonal antibodies.

[0082] Example 5: Preparation of paroxetine polyclonal antibodies:

[0083] Paroxetine polyclonal antibodies were prepared using the paroxetine antigen obtained in Example 2, and the rest of the preparation process was the same as in Example 4 to obtain paroxetine polyclonal antibodies.

[0084] Example 6: Preparation of paroxetine polyclonal antibodies:

[0085] Paroxetine polyclonal antibodies were prepared using the paroxetine antigen obtained in Example 3, and the rest of the preparation process was the same as in Example 4 to obtain paroxetine polyclonal antibodies.

[0086] Example 7: Preparation of paroxetine colloidal gold test paper:

[0087] A. Test line and quality control line: 0.2 mg / mL of the paroxetine antigen obtained in Example 1 was sprayed on a nitrocellulose membrane at a spray volume of 1.0 μL / cm as a test line, and 1.0 mg / mL of goat anti-rabbit IgG and 0.1 mg / mL of goat anti-mouse IgG were sprayed on a nitrocellulose membrane at a spray volume of 1.0 μL / cm as a quality control line;

[0088] B. Paroxetine polyclonal antibody-colloidal gold complex: the paroxetine polyclonal antibody obtained in Example 4 was dissolved in a colloidal gold solution to obtain a paroxetine polyclonal antibody solution, the pH value of the colloidal gold solution was adjusted to 7.6, 1 mL of the paroxetine polyclonal antibody solution was added, 10 mL of 1% PEG2000 buffer was added after stirring, and the mixture was centrifuged at 100000 g for 60 min at 4° C. to remove the precipitate and resuspend in a buffer containing PEG2000, and the mixture was repeated 3 times to obtain a colloidal gold labeled protein, and then the colloidal gold labeled protein was purified by gel filtration, eluted with a PBS buffer containing 1% BSA and 0.02% sodium azide, and the paroxetine polyclonal antibody-colloidal gold complex was obtained;

[0089] C. Conjugation pad: dilute the paroxetine polyclonal antibody-colloidal gold complex in PBS buffer containing 1% BSA, 50 mg / mL trehalose and 200 mg / mL sucrose to 1 / 3 of the initial OD value of the complex, and spray it evenly on the conjugation pad at a spray volume of 1.0 μL / cm;

[0090] D. Assembly: Assemble the film-doped sheet base plate with the binding pad, sample pad, and absorbent paper into a large card, and cut it into strips to obtain the paroxetine colloidal gold test strips.

[0091] Example 8: Preparation of paroxetine colloidal gold test paper:

[0092] The paroxetine antigen is the paroxetine antigen obtained in Example 2, the paroxetine polyclonal antibody is the paroxetine polyclonal antibody obtained in Example 5, and the rest of the preparation process is the same as that in Example 7.

[0093] Example 9: Preparation of paroxetine colloidal gold test paper:

[0094] The paroxetine antigen is the paroxetine antigen obtained in Example 3, the paroxetine monoclonal antibody is the paroxetine monoclonal antibody obtained in Example 6, and the rest of the preparation process is the same as that in Example 7.

[0095] Paroxetine solutions of different concentrations were prepared, and the paroxetine colloidal gold detection reagent strip prepared in Example 7 was used to perform functional detection on the samples. The color intensity of the detection line was determined using a colloidal gold colorimetric card. Each concentration was measured 5 times and the average value was taken. The results are shown in Table 1.

[0096] Table 1. Color intensity of paroxetine colloidal gold test paper

[0097] Sample concentration (ng / mL) T-line strength 0 G9 250 G5 500 G3 750 G2 1000 G1 1500 G1

[0098] The result of the colloidal gold test paper is interpreted by the colorimetric card method, and G1-G10 represents the color development degree of the T-line colloidal gold, wherein G1 is a T-line without color development under naked eye observation, indicating a strong positive; G10 is a T-line with a dark color, indicating a strong negative. As can be seen from Table 1, the detection concentration of the test paper prepared by the present invention can be as low as 500ng / mL, with high sensitivity, and the results can be obtained by naked eye observation. The paroxetine colloidal gold detection reagent strips prepared in Examples 7-9 were used to perform functional tests on 101 clinical urine samples. Among the clinical urine samples, 70 were negative and 31 were positive, and the paroxetine concentration was distributed between 500 and 1500ng / mL. The detection results are shown in Table 2.

[0099] Table 2. Clinical sample test results

[0100]

[0101] It can be seen from Table 2 that the paroxetine colloidal gold test paper of the present invention has a high detection accuracy rate for clinical samples, and the overall accuracy is higher than 90%.

Claims

1. A paroxetine hapten, characterized in that Its molecular structure is shown in formula (I): 。 2. A method for preparing the paroxetine hapten according to claim 1, characterized in that The method comprises the following steps: Step (1), dissolving 2-hydroxyparoxetine in N,N-dimethylformamide, adding triethylamine and di-tert-butyl dicarbonate, heating under reflux at 45-50° C., cooling the reaction solution to room temperature after the reaction is complete, and evaporating the solvent under reduced pressure to obtain an oily substance A; Step (2), dissolving the oil phase substance A in pyridine, then adding glutaric anhydride and 4-dimethylaminopyridine, heating under reflux at 98-100° C., cooling the reaction solution to room temperature after the reaction is complete, and evaporating the solvent under reduced pressure to obtain an oily residue; adding dichloromethane to the oily residue, washing with purified water and saturated NaCl respectively, collecting the dichloromethane phase, drying, filtering, and evaporating under reduced pressure to obtain an oily substance B; Step (3), dissolving the oil B in a mixed solution of HCl and 1,4-dioxane, heating with stirring under reflux at 98-100° C., and evaporating the solvent under reduced pressure to obtain an oil C; Step (4), dissolving the oily substance C in benzene, placing in an ice-water bath, adding trifluoroacetic anhydride, stirring at room temperature for a period of time, and then heating and stirring at 78-81° C. and reflux. After the reaction is complete, the reaction solution is dried and separated to obtain the oily substance D, i.e., the artificial hapten of paroxetine.

3. The method according to claim 2, characterized in that In step (1), the molar ratio of 2-hydroxyparoxetine to triethylamine and di-tert-butyl dicarbonate is 1:2:2; in step (2), the molar ratio of oily substance A, glutaric anhydride and 4-dimethylaminopyridine (DMAP) is (10-12):(4-5.5):(1-2); in step (4), the molar ratio of oily substance C to trifluoroacetic anhydride is (1-1.3):(1.8-2.4).

4. A paroxetine artificial antigen, characterized in that The paroxetine hapten according to claim 1 is coupled with a carrier protein; Its molecular structure is shown in formula (II): Wherein R represents carrier protein.

5. The paroxetine artificial antigen according to claim 4, characterized in that The carrier protein is one of bovine serum albumin, bovine gamma globulin, bovine thyroglobulin, keyhole limpet hemocyanin and chicken ovalbumin.

6. A method for preparing the paroxetine artificial antigen according to claim 4 or 5, characterized in that The method comprises the following steps: Step (1), dissolving the paroxetine artificial hapten in N,N-dimethylformamide DMF, adding N-hydroxysuccinimide NHS and cyclohexylcarbodiimide, reacting at room temperature with stirring for 15 to 18 hours, and centrifuging to obtain the supernatant to obtain liquid A; Step (2), dissolving the carrier protein in PBS buffer as liquid B, wherein the concentration of the carrier protein in liquid B is 4-9 mg / ml; Step (3), adding liquid A dropwise to liquid B under stirring, and standing overnight at low temperature to obtain an artificial antigen mixture; Step (4), dialyzing the artificial antigen mixture obtained in step (3) in alkaline dialysate for multiple times, and then transferring it into PBS buffer for multiple dialysates. After the dialysis is completed, the supernatant is centrifuged to obtain the paroxetine artificial antigen.

7. The method according to claim 6, characterized in that In step (1), the molar ratio of the paroxetine artificial hapten, N-hydroxysuccinimide and cyclohexylcarbodiimide is (1.3-1.4):1.9:1.

9.

8. The method according to claim 7, characterized in that In step (3), the volume ratio of solution A to solution B is 1:

5.

9. A paroxetine colloidal gold test paper, comprising a test line, a quality control line, and a binding pad; characterized in that The test line is obtained by using the paroxetine artificial antigen described in claim 4 or 5 as a raw material for film deposition.

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