Method for determining contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil

Abstract

The invention provides a method for determining the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil, and belongs to the technical field of detection methods. Comprising the following steps: (1) preparing an isotope internal standard working solution; (2) adding an adsorbent into the opium poppy oil sample, stirring for adsorption, filtering, adding a solid into a solvent, heating for desorption, filtering, washing, combining a washing solution and a filtrate, adding an isotope internal standard working solution, filtering, and fixing the volume to obtain a test solution; (3) preparing a control stock solution; (4) preparing a reference substance solution; and (5) determining the contents of codeine, morphine, papaverine, narcotine and thebaine in the opium poppy oil by adopting LC-MS / MS (Liquid Chromatography-Mass Spectrometry / Mass Spectrometry). The simple and effective method is adopted for determining the contents of codeine, morphine, papaverine, narcotine and thebaine in the opium poppy oil, the linear range is large, the detection limit is low, the accuracy rate is high, the sensitivity is high, the linearity is good, and the method is a reliable and effective detection method.

Description

Technical Field

[0001] The invention relates to the technical field of detection methods, and in particular to a method for determining the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil. Background Art

[0002] Poppy seed oil is a vegetable oil extracted from the seeds of the poppy plant (Papaver somniferum L.). Although poppy seeds themselves are generally considered to contain no or only trace amounts of alkaloids, the surface of the seeds can be easily contaminated by the sap (opium) of the poppy capsule (which contains high concentrations of alkaloids) during cultivation, harvesting, processing, and storage. Furthermore, studies have shown that under certain conditions (such as mechanical damage, pests and diseases, and specific varieties), alkaloids can enter the seed internally through endogenous pathways. Therefore, poppy seed oil may contain a variety of natural alkaloids derived from contamination or endogenous accumulation. The five most important and pharmacologically and toxicologically significant are: morphine, codeine, papaverine, noscapine, and thebaine. To prevent health risks, abuse, and drug production, the levels of these five substances in poppy seed oil need to be monitored.

[0003] Poppy oil itself is a high-fat matrix, containing large amounts of lipid substances such as triglycerides and free fatty acids. These substances can seriously interfere with subsequent chromatographic separation and mass spectrometry detection, leading to column contamination, poor peak shape, and decreased sensitivity. Traditional methods for water-soluble samples or plant extracts are generally not applicable. In addition, these five alkaloids differ significantly in their chemical structure, polarity, acidity and alkalinity (morphine and codeine have phenolic hydroxyl groups and are weakly acidic; papaverine, narcotine, and thebaine are tertiary amines and are weakly alkaline), and solubility (different solubility in oil matrices and water). Developing a method that can simultaneously and effectively extract, purify, and separate these five compounds with very different properties is a core challenge. Summary of the Invention

[0004] The present invention aims to provide a method for determining the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil. The method adopts a simple and effective method to determine the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil. The method has a large linear range, a low detection limit, high accuracy, high sensitivity, a regression coefficient (r) of ≥0.990 and good linearity, and is a reliable and effective detection method.

[0005] The technical solution of the present invention is achieved as follows:

[0006] The present invention provides a method for determining the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil, comprising the following steps:

[0007] (1) Preparation of isotope internal standard working solutions: Accurately pipette morphine-D3 isotope solution and codeine-D3 isotope solution into volumetric flasks, and adjust the volume to prepare the internal standard working solutions of morphine-D3 and codeine-D3 respectively;

[0008] (2) Preparation of test solution: add adsorbent to poppy oil sample, stir for adsorption, filter, add solid to solvent, heat for desorption, filter, wash, combine washing solution and filtrate, add isotope internal standard working solution, filter, and adjust to volume to prepare test solution;

[0009] (3) Preparation of control stock solutions: Prepare corresponding stock solutions using codeine, morphine, papaverine, thebaine, and narcotine standards;

[0010] (4) Preparation of reference solution: dilute the reference stock solution to prepare the reference solution;

[0011] (5) Detection: LC-MS / MS was used to determine the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil.

[0012] This detection method first uses liquid chromatography (HPLC) to separate the detection target in the test sample; the separated sample is ionized into charged ions in the ion source; then, the first-stage mass spectrometer screens the target parent ion based on the mass-to-charge ratio (m / z) and eliminates other interfering ions; after the parent ion enters the collision chamber, it fragments into daughter ions through collision-induced dissociation (CID); finally, the second-stage mass spectrometer detects the daughter ions, generating a fragment spectrum that reveals the chemical structure characteristics of the parent ion.

[0013] As a further improvement of the present invention, the adsorbent includes adsorbent A, adsorbent B and adsorbent C in a mass ratio of 5-7:4-6:1-3;

[0014] The preparation method of the adsorbent A is as follows:

[0015] Morphine, thebaine, monomer, cross-linking agent, emulsifier and porogen are mixed and added to ethyl acetate, mixed evenly, added dropwise to water, emulsified, and under inert gas protection, an initiator is added, heated and stirred to react, the product is filtered, added to a Soxhlet extractor, extracted with a mixed solvent until no morphine and thebaine are detected, washed, and dried to prepare adsorbent A;

[0016] The preparation method of the adsorbent B is as follows:

[0017] S1. Preparation of a chiral amino acid-coupled MOF: L-aspartic acid and L-glutamic acid were added to water, followed by N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and activated by stirring. UiO-66-NH2 was added, the reaction was stirred, filtered, washed, and dried to obtain a chiral amino acid-coupled MOF.

[0018] S2. Double bond grafting modification: Maleic acid was added to water, followed by N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and the mixture was stirred for activation. A chiral amino acid was added to couple the MOF, and the mixture was stirred for reaction. The mixture was filtered, washed, and dried to obtain the modified chiral amino acid-coupled MOF.

[0019] S3. Copolymerization: The modified chiral amino acid-coupled MOF, methyl methacrylate, styrene, and N-isopropylacrylamide were mixed in ethyl acetate, uniformly mixed, and dropwise added to water for emulsification. Under inert gas protection, an initiator was added, and the reaction was heated with stirring. The product was filtered to produce adsorbent B.

[0020] The preparation method of the adsorbent C is as follows:

[0021] T1 Preparation of diazonium sulfanilate solution: sulfanilic acid and sodium nitrite were dissolved in lye, heated and mixed, cooled to room temperature, hydrochloric acid was added dropwise, and the reaction was stirred to obtain a diazonium sulfanilate solution;

[0022] T2. Preparation of sulfonated / double-bond co-modified graphene oxide: Maleic acid was added to toluene, graphene oxide and p-toluenesulfonic acid were added, the mixture was heated to reflux with stirring, the reaction was centrifuged, washed, and dried, and the product was added to a solution of diazonium p-aminobenzenesulfonic acid, the reaction was stirred, centrifuged, washed, and dried to obtain sulfonated / double-bond co-modified graphene oxide;

[0023] T3. Copolymerization: Sulfonated / double-bond co-modified graphene oxide, methyl methacrylate, styrene, and N-isopropylacrylamide were mixed in ethyl acetate, uniformly mixed, and dropwise added to water for emulsification. Under inert gas protection, an initiator was added, and the mixture was heated and stirred for reaction. The product was filtered to obtain adsorbent C.

[0024] As a further improvement of the present invention, in the preparation process of adsorbent A, the monomers include methyl methacrylate, styrene, 1-vinyl-3-(aminopropyl) imidazole bis(trifluoromethanesulfonyl) imide salt, and N-isopropyl acrylamide in a mass ratio of 3-5:4-6:1-2:5-7, the cross-linking agent is ethylene glycol dimethacrylate, the emulsifier is a mixture of Tween-80 and Span-80 in a mass ratio of 2-3:3-5, and the porogen is hexadecyltrimethylammonium chloride or hexadecyltrimethylammonium bromide. ammonium, the initiator is selected from at least one of azobisisobutyronitrile, sodium persulfate, potassium persulfate, and ammonium persulfate, the temperature of the heating and stirring reaction is 55-65° C., and the time is 4-6 hours. The mass ratio of morphine, thebaine, monomer, cross-linking agent, emulsifier, porogen and initiator is 0.5-1:0.5-1:8-12:0.2-0.4:0.5-1:0.1-0.2:0.01-0.15, and the mixed solvent is a mixed solvent of acetonitrile and dichloromethane, and the volume ratio is 1:1-3.

[0025] As a further improvement of the present invention, the mass ratio of L-aspartic acid, L-glutamic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and UiO-66-NH2 in step S1 is 2-3:1-2:2-4:2-4:10; the mass ratio of maleic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and chiral amino acid coupling MOF in step S2 is 2-4:1-2:1-2:10; the mass ratio of the modified chiral amino acid coupling MOF, methyl methacrylate, styrene, N-isopropylacrylamide, and initiator in step S3 is 5-7:4-6:3-5:6-8:0.1-0.2, the temperature of the heating and stirring reaction is 60-70°C, the time is 3-5h, and the initiator is selected from at least one of sodium persulfate, potassium persulfate, and ammonium persulfate.

[0026] As a further improvement of the present invention, the mass ratio of p-aminobenzenesulfonic acid, sodium nitrite, and hydrochloric acid in step T1 is 0.04-0.06:1:6-8, and the stirring reaction time is 3-5 hours; the mass ratio of maleic acid, graphene oxide, p-toluenesulfonic acid, and p-aminobenzenesulfonic acid diazonium salt solution in step T2 is 2-3:8-10:0.5-1:15-20, and the heating reflux stirring reaction time is 4-7 hours, and the stirring reaction time is 10-12 hours; the mass ratio of sulfonated / double bond co-modified graphene oxide, methyl methacrylate, styrene, N-isopropylacrylamide, and initiator in step T3 is 4-6:3-5:3-4:5-7:0.1-0.2, and the heating stirring reaction time is 55-65°C for 4-6 hours.

[0027] As a further improvement of the present invention, the chromatographic conditions for the detection are as follows: chromatographic column: Agilent ZORBAXSB-Aq 3.5μm 3.0×100mm, column temperature 35-45°C, injection plate temperature 3-7°C, mobile phase: A: 2-7Mmol / L ammonium acetate + 0.05-0.15% formic acid (v / v); B: 0.05-0.15% formic acid acetonitrile solution (v / v), flow rate 0.1-0.5mL / min, needle wash solution MeOH:H2O=1:1 (v:v).

[0028] As a further improvement of the present invention, the mass spectrometry conditions for the detection are: ion source ESI, curtain gas 30-40psi, temperature 550-650°C, ionization voltage 5000-6000V, spray gas 45-55psi, auxiliary heating gas 45-55psi, and scanning mode positive ion mode.

[0029] As a further improvement of the present invention, the linear range of codeine is 62.5 ng / mL-187.5 ng / mL, the linear range of morphine is 62.5 ng / mL-187.5 ng / mL, the linear range of papaverine is 12.5 ng / mL-37.5 ng / mL, the linear range of narcotine is 12.5 ng / mL-37.5 ng / mL, and the linear range of thebaine is 12.5 ng / mL-37.5 ng / mL.

[0030] As a further improvement of the present invention, the regression coefficient of codeine is 0.995, the regression coefficient of morphine is 0.994, the regression coefficient of papaverine is 0.995, the regression coefficient of narcotine is 0.998, and the regression coefficient of thebaine is 0.999.

[0031] As a further improvement of the present invention, the mass ratio of the poppy oil, the adsorbent, and the isotope internal standard working solution is 15-20:1-2:0.000001-0.000002, the solvent is a mixed solution of formic acid and acetonitrile in a volume ratio of 0.5-1:100, and the heating desorption temperature is 40-45° C. and the time is 20-40 minutes.

[0032] The present invention has the following beneficial effects:

[0033] The invention adopts a simple and effective method to determine the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil, has a large linear range, a low detection limit, high accuracy, high sensitivity, a regression coefficient (r) of ≥0.990 and good linearity, and is a reliable and effective detection method.

[0034] The adsorbent A, adsorbent B and adsorbent C prepared by the present invention have good adsorption properties for alkaloid substances in poppy oil and fast desorption rates. The adsorbents can be reused multiple times, thereby reducing costs.

[0035] Among them, adsorbent A has a good adsorption effect on morphine and thebaine, which have similar structures. It adopts double template co-imprinting to simultaneously imprint morphine and thebaine, effectively adsorbing and separating morphine and thebaine. At the same time, the polymer also contains a thermosensitive monomer, which has temperature controllable recognition of the template molecules morphine and thebaine. At low temperatures, the volume swells, which helps the adsorption of morphine and thebaine. When the temperature is increased, the volume shrinks, and the morphine and thebaine molecules are desorbed, thereby achieving efficient separation of the molecules through the change in the affinity of the imprinted cavity binding to the template molecule. The polymer also contains an ionic liquid monomer with an imidazole group, which uses π-π stacking and hydrogen bond adsorption to have a specific affinity for the indole structure of morphine and thebaine, thereby improving the adsorption rate of adsorbent A for morphine and thebaine.

[0036] Adsorbent B couples chiral amino acids to the surface of the MOF material UiO-66-NH2 via aminocarboxyl condensation, achieving specific adsorption of codeine through chiral cavity induction and electrostatic complementation. The pore size of the MOFs matches the molecular size of codeine. Codeine, a tertiary amine alkaloid, binds tightly to negatively charged amino acid side chains through directional hydrogen bonds, resulting in a high adsorption capacity. Simultaneously, the chiral amino acid-coupled MOF is coupled with maleic acid, resulting in a modified chiral amino acid-coupled MOF with double bonds on its surface, enabling copolymerization with monomers. The thermosensitive monomers, through the imprinted cavity, change their affinity for binding to codeine molecules, enabling molecular adsorption and desorption.

[0037] Adsorbent C couples sulfonic acid groups and double bonds on the surface of graphene oxide materials, achieving efficient adsorption of papaverine and narcotine through sulfonic acid group ion exchange and π-π stacking. The sulfonic acid group (-SO3H) of GO-SO3H forms a strong electrostatic interaction with the benzylisoquinoline structure of papaverine and the isoquinoline structure of narcotine, thereby accelerating separation and improving separation efficiency. In addition, the double bond can copolymerize with the monomer. The thermosensitive monomer in it changes the affinity of the imprinted cavity to bind to the papaverine and narcotine molecules, thereby achieving molecular adsorption and desorption separation. DETAILED DESCRIPTION

[0038] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0039] Preparation Example 1 Adsorbent A

[0040] The preparation method is as follows:

[0041] 0.5 g of morphine, 0.5 g of thebaine, 8 g of monomer, 0.2 g of ethylene glycol dimethacrylate, 0.5 g of emulsifier and 0.1 g of hexadecyltrimethylammonium chloride were mixed and added to 200 mL of ethyl acetate, stirred and mixed for 15 minutes, and then added dropwise to 500 mL of water. The mixture was emulsified at 5000 rpm for 15 minutes. Under nitrogen protection, 0.01 g of potassium persulfate was added, and the mixture was heated to 55° C. and stirred for 4 hours. The product was filtered and added to a Soxhlet extractor. It was extracted with a mixed solvent until no morphine and thebaine were detected. The mixture was washed and dried to prepare adsorbent A.

[0042] The mixed solvent is a mixed solvent of acetonitrile and dichloromethane, with a volume ratio of 1:1;

[0043] The emulsifier is a mixture of Tween-80 and Span-80, with a mass ratio of 2:3;

[0044] The monomers include methyl methacrylate, styrene, 1-vinyl-3-(aminopropyl) imidazole bis(trifluoromethanesulfonyl) imide salt, and N-isopropylacrylamide, with a mass ratio of 3:4:1:5.

[0045] Preparation Example 2 Adsorbent A

[0046] The preparation method is as follows:

[0047] 1 g of morphine, 1 g of thebaine, 12 g of monomer, 0.4 g of ethylene glycol dimethacrylate, 1 g of emulsifier, and 0.2 g of hexadecyltrimethylammonium chloride were mixed and added to 200 mL of ethyl acetate, stirred for 15 minutes, and then added dropwise to 500 mL of water. The mixture was emulsified at 5000 rpm for 15 minutes. Under nitrogen protection, 0.15 g of sodium persulfate was added, and the mixture was heated to 65° C. and stirred for 6 hours. The product was filtered and added to a Soxhlet extractor. It was extracted with a mixed solvent until no morphine and thebaine were detected. The mixture was washed and dried to obtain adsorbent A.

[0048] The mixed solvent is a mixed solvent of acetonitrile and dichloromethane, with a volume ratio of 1:3;

[0049] The emulsifier is a mixture of Tween-80 and Span-80, with a mass ratio of 3:5;

[0050] The monomers include methyl methacrylate, styrene, 1-vinyl-3-(aminopropyl) imidazole bis(trifluoromethanesulfonyl) imide salt, and N-isopropylacrylamide, with a mass ratio of 5:6:2:7.

[0051] Preparation Example 3 Adsorbent A

[0052] The preparation method is as follows:

[0053] 0.7 g of morphine, 0.7 g of thebaine, 10 g of monomer, 0.3 g of ethylene glycol dimethacrylate, 0.7 g of emulsifier and 0.15 g of hexadecyltrimethylammonium bromide were mixed and added to 200 mL of ethyl acetate, stirred and mixed for 15 minutes, and then added dropwise to 500 mL of water. The mixture was emulsified at 5000 rpm for 15 minutes. Under nitrogen protection, 0.012 g of azobisisobutyronitrile was added, and the mixture was heated to 60° C. and stirred for 5 hours. The product was filtered and added to a Soxhlet extractor. It was extracted with a mixed solvent until no morphine and thebaine were detected. The mixture was washed and dried to prepare adsorbent A.

[0054] The mixed solvent is a mixed solvent of acetonitrile and dichloromethane, with a volume ratio of 1:2;

[0055] The emulsifier is a mixture of Tween-80 and Span-80, with a mass ratio of 2.5:4;

[0056] The monomers include methyl methacrylate, styrene, 1-vinyl-3-(aminopropyl) imidazole bis(trifluoromethanesulfonyl) imide salt, and N-isopropylacrylamide, with a mass ratio of 4:5:1.5:6.

[0057] Comparative Preparation Example 1

[0058] Compared with Preparation Example 3, the difference is that 1-vinyl-3-(aminopropyl) imidazole bis(trifluoromethanesulfonyl) imide salt is not added. The monomers include methyl methacrylate, styrene, and N-isopropylacrylamide in a mass ratio of 4:6.5:6.

[0059] Comparative Preparation Example 2

[0060] Compared with Preparation Example 3, the difference is that N-isopropylacrylamide is not added. The monomers include methyl methacrylate, styrene, 1-vinyl-3-(aminopropyl)imidazolium bis(trifluoromethanesulfonyl)imide salt, and N-isopropylacrylamide in a mass ratio of 4:11:1.5.

[0061] Preparation Example 4 Adsorbent B

[0062] The preparation method is as follows:

[0063] S1. Preparation of a chiral amino acid-coupled MOF: 2 g of L-aspartic acid and 1 g of L-glutamic acid were added to 200 mL of water, along with 2 g of N-hydroxysuccinimide and 2 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of UiO-66-NH2 was added and the mixture was stirred for 7 h. The mixture was filtered, washed, and dried to obtain a chiral amino acid-coupled MOF.

[0064] S2. Double bond grafting modification: 2 g of maleic acid was added to 200 mL of water, along with 1 g of N-hydroxysuccinimide and 1 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of chiral amino acid-coupled MOF was added and stirred for 5 h. The mixture was filtered, washed, and dried to obtain the modified chiral amino acid-coupled MOF.

[0065] S3. Copolymerization: 5 g of the modified chiral amino acid-coupled MOF, 4 g of methyl methacrylate, 3 g of styrene, and 6 g of N-isopropylacrylamide were added to 200 mL of ethyl acetate and stirred for 10 min. The mixture was then added dropwise to 300 mL of water and emulsified at 7000 rpm for 15 min. Under nitrogen, 0.1 g of sodium persulfate was added, and the mixture was heated to 60°C and stirred for 3 h. The product was filtered to obtain adsorbent B.

[0066] Preparation Example 5 Adsorbent B

[0067] The preparation method is as follows:

[0068] S1. Preparation of a chiral amino acid-coupled MOF: 3 g L-aspartic acid and 2 g L-glutamic acid were added to 200 mL of water, along with 4 g N-hydroxysuccinimide and 4 g 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g UiO-66-NH2 was added and the mixture was stirred for 10 h. The mixture was filtered, washed, and dried to obtain a chiral amino acid-coupled MOF.

[0069] S2. Double bond grafting modification: 4 g of maleic acid was added to 200 mL of water, along with 2 g of N-hydroxysuccinimide and 2 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of chiral amino acid-coupled MOF was added and the reaction was stirred for 7 h. The mixture was filtered, washed, and dried to obtain the modified chiral amino acid-coupled MOF.

[0070] S3. Copolymerization: 7 g of the modified chiral amino acid-coupled MOF, 6 g of methyl methacrylate, 5 g of styrene, and 8 g of N-isopropylacrylamide were added to 200 mL of ethyl acetate and stirred for 10 min. The mixture was then added dropwise to 300 mL of water and emulsified at 7000 rpm for 15 min. Under nitrogen, 0.2 g of potassium persulfate was added, and the mixture was heated to 70°C and stirred for 5 h. The product was filtered to obtain adsorbent B.

[0071] Preparation Example 6 Adsorbent B

[0072] The preparation method is as follows:

[0073] S1. Preparation of a chiral amino acid-coupled MOF: 2.5 g of L-aspartic acid and 1.5 g of L-glutamic acid were added to 200 mL of water, along with 3 g of N-hydroxysuccinimide and 3 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of UiO-66-NH2 was added and the mixture was stirred for 8 h. The mixture was filtered, washed, and dried to obtain a chiral amino acid-coupled MOF.

[0074] S2. Double bond grafting modification: 3 g of maleic acid was added to 200 mL of water, along with 1.5 g of N-hydroxysuccinimide and 1.5 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred for 30 min, and then 10 g of chiral amino acid-coupled MOF was added. The reaction was stirred for 6 h, filtered, washed, and dried to obtain the modified chiral amino acid-coupled MOF.

[0075] S3. Copolymerization: 6 g of the modified chiral amino acid-coupled MOF, 5 g of methyl methacrylate, 4 g of styrene, and 7 g of N-isopropylacrylamide were added to 200 mL of ethyl acetate and stirred for 10 minutes. The mixture was then added dropwise to 300 mL of water and emulsified at 7000 rpm for 15 minutes. Under nitrogen, 0.15 g of ammonium persulfate was added, and the mixture was heated to 65°C and stirred for 4 hours. The product was filtered to obtain adsorbent B.

[0076] Comparative Preparation Example 3

[0077] Compared with Preparation Example 3, the difference is that L-aspartic acid is not added in Step S1.

[0078] The details are as follows:

[0079] S1. Preparation of a chiral amino acid-coupled MOF: 4 g of L-glutamic acid was added to 200 mL of water, along with 3 g of N-hydroxysuccinimide and 3 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of UiO-66-NH2 was added and the mixture was stirred for 8 h. The mixture was filtered, washed, and dried to obtain a chiral amino acid-coupled MOF.

[0080] Comparative Preparation Example 4

[0081] Compared with Preparation Example 3, the difference is that L-glutamic acid is not added in step S1.

[0082] The details are as follows:

[0083] S1. Preparation of a chiral amino acid-coupled MOF: 4 g of L-aspartic acid was added to 200 mL of water, along with 3 g of N-hydroxysuccinimide and 3 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of UiO-66-NH2 was added and the mixture was stirred for 8 h. The mixture was filtered, washed, and dried to obtain a chiral amino acid-coupled MOF.

[0084] Comparative Preparation Example 5

[0085] Compared with Preparation Example 3, the difference is that step S1 is not performed.

[0086] The details are as follows:

[0087] S1. Double bond grafting modification: 3 g of maleic acid was added to 200 mL of water, along with 1.5 g of N-hydroxysuccinimide and 1.5 g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride. The mixture was stirred and activated for 30 min. 10 g of UiO-66-NH2 was added and the reaction was stirred for 6 h. The mixture was filtered, washed, and dried to obtain the modified MOF.

[0088] S2. Copolymerization: 6 g of modified MOF, 5 g of methyl methacrylate, 4 g of styrene, and 7 g of N-isopropylacrylamide were added to 200 mL of ethyl acetate and stirred for 10 minutes. The mixture was then added dropwise to 300 mL of water and emulsified at 7000 rpm for 15 minutes. Under nitrogen, 0.15 g of ammonium persulfate was added, and the mixture was heated to 65°C and stirred for 4 hours. The product was filtered to obtain adsorbent B.

[0089] Preparation Example 7 Adsorbent C

[0090] The preparation method is as follows:

[0091] T1. Preparation of diazonium sulfanilate solution: Dissolve 0.04 g of sulfanilic acid and 1 g of sodium nitrite in 20 mL of 2 wt% NaOH solution, heat to 50°C, stir for 15 minutes, cool to room temperature, add 6 g of 36 wt% hydrochloric acid dropwise, and stir for 3 hours to obtain a diazonium sulfanilate solution.

[0092] T2. Preparation of sulfonated / double-bond co-modified graphene oxide: 2 g of maleic acid was added to 200 mL of toluene, followed by 8 g of graphene oxide and 0.5 g of p-toluenesulfonic acid. The mixture was stirred under reflux for 4 h, centrifuged, washed, and dried. The product was then added to 15 g of diazonium p-aminobenzenesulfonic acid solution, stirred for 10 h, centrifuged, washed, and dried to obtain sulfonated / double-bond co-modified graphene oxide.

[0093] T3. Copolymerization: 4 g of sulfonated / double-bond co-modified graphene oxide, 3 g of methyl methacrylate, 3 g of styrene, and 5 g of N-isopropylacrylamide were mixed in 200 mL of ethyl acetate and stirred for 20 min. The mixture was then added dropwise to 300 mL of water and emulsified at 8000 rpm for 15 min. Under nitrogen protection, 0.1 g of sodium persulfate was added, and the mixture was heated to 55°C and stirred for 4 h. The product was filtered to obtain adsorbent C.

[0094] Preparation Example 8 Adsorbent C

[0095] The preparation method is as follows:

[0096] T1. Preparation of diazonium sulfanilate solution: Dissolve 0.06 g of sulfanilic acid and 1 g of sodium nitrite in 20 mL of 2 wt% NaOH solution, heat to 50°C, stir for 15 minutes, cool to room temperature, add 8 g of 36 wt% hydrochloric acid dropwise, and stir for 5 hours to obtain a diazonium sulfanilate solution.

[0097] T2. Preparation of sulfonated / double-bond co-modified graphene oxide: 3 g of maleic acid was added to 200 mL of toluene, followed by 10 g of graphene oxide and 1 g of p-toluenesulfonic acid. The mixture was stirred under reflux for 7 h, centrifuged, washed, and dried. The product was then added to 20 g of diazonium p-aminobenzenesulfonic acid solution, stirred for 12 h, centrifuged, washed, and dried to obtain sulfonated / double-bond co-modified graphene oxide.

[0098] T3. Copolymerization: 6 g of sulfonated / double-bond co-modified graphene oxide, 5 g of methyl methacrylate, 4 g of styrene, and 7 g of N-isopropylacrylamide were added to 200 mL of ethyl acetate and stirred for 20 min. The mixture was then added dropwise to 300 mL of water and emulsified at 8000 rpm for 15 min. Under nitrogen protection, 0.2 g of ammonium persulfate was added, and the mixture was heated to 65°C and stirred for 6 h. The product was filtered to obtain adsorbent C.

[0099] Preparation Example 9 Adsorbent C

[0100] The preparation method is as follows:

[0101] T1. Preparation of diazonium sulfanilate solution: Dissolve 0.05 g of sulfanilic acid and 1 g of sodium nitrite in 20 mL of 2 wt% NaOH solution, heat to 50°C, stir for 15 minutes, cool to room temperature, add 7 g of 36 wt% hydrochloric acid dropwise, and stir for 4 hours to obtain a diazonium sulfanilate solution.

[0102] T2. Preparation of sulfonated / double-bond co-modified graphene oxide: 2.5 g of maleic acid was added to 200 mL of toluene, followed by 9 g of graphene oxide and 0.7 g of p-toluenesulfonic acid. The mixture was stirred at reflux for 5.5 h, centrifuged, washed, and dried. The product was then added to 17 g of p-aminobenzenesulfonic acid diazonium salt solution, stirred for 11 h, centrifuged, washed, and dried to obtain sulfonated / double-bond co-modified graphene oxide.

[0103] T3. Copolymerization: 5 g of sulfonated / double-bond co-modified graphene oxide, 4 g of methyl methacrylate, 3.5 g of styrene, and 6 g of N-isopropylacrylamide were mixed in 200 mL of ethyl acetate and stirred for 20 min. The mixture was then added dropwise to 300 mL of water and emulsified at 8000 rpm for 15 min. Under nitrogen protection, 0.15 g of potassium persulfate was added, and the mixture was heated to 60°C and stirred for 5 h. The product was filtered to obtain adsorbent C.

[0104] Comparative Preparation Example 6

[0105] Compared with Preparation Example 9, the difference is that sulfonation is not performed in Step T2.

[0106] The details are as follows:

[0107] T1. Preparation of double bond-modified graphene oxide: 2.5 g of maleic acid was added to 200 mL of toluene, 9 g of graphene oxide and 0.7 g of p-toluenesulfonic acid were added, and the mixture was stirred under reflux for 5.5 h, centrifuged, washed, and dried to obtain double bond-modified graphene oxide;

[0108] T2. Copolymerization: 5 g of double-bond-modified graphene oxide, 4 g of methyl methacrylate, 3.5 g of styrene, and 6 g of N-isopropylacrylamide were mixed in 200 mL of ethyl acetate and stirred for 20 min. The mixture was then added dropwise to 300 mL of water and emulsified at 8000 rpm for 15 min. Under nitrogen protection, 0.15 g of potassium persulfate was added, and the mixture was heated to 60°C and stirred for 5 h. The product was filtered to obtain adsorbent C.

[0109] Comparative Preparation Example 7

[0110] Compared with Preparation Example 9, the difference is that N-isopropylacrylamide is not added in Step T3.

[0111] The details are as follows:

[0112] T3. Copolymerization: 5 g of sulfonated / double-bond co-modified graphene oxide, 4 g of methyl methacrylate, and 9.5 g of styrene were added to 200 mL of ethyl acetate and stirred for 20 min. The mixture was then added dropwise to 300 mL of water and emulsified at 8000 rpm for 15 min. Under nitrogen protection, 0.15 g of potassium persulfate was added, and the mixture was heated to 60°C and stirred for 5 h. The product was filtered to obtain adsorbent C.

[0113] Example 1

[0114] This embodiment provides a method for determining the content of codeine, morphine, papaverine, narcotine, and thebaine in poppy oil, comprising the following steps:

[0115] (1) Preparation of isotope internal standard working solutions: Accurately pipette appropriate amounts of morphine-D3 isotope solution and codeine-D3 isotope solution into volumetric flasks, and dilute to the mark with water to prepare internal standard working solutions with a concentration of 5.0 μg / mL for both morphine-D3 and codeine-D3.

[0116] (2) Preparation of test solution: Add 1 g of adsorbent to 20 g of poppy oil sample, stir and adsorb for 20 min, filter, add the solid to the solvent, heat to 40 ° C, stir and desorb for 20 min, filter, wash, combine the washing liquid and the filtrate, add 2 μg of isotope internal standard working solution, filter through a 0.22 μm filter membrane, and adjust the volume to prepare the test solution;

[0117] The solvent is a mixed solution of formic acid and acetonitrile with a volume ratio of 0.5:100;

[0118] The adsorbent includes adsorbent A prepared in Preparation Example 1, adsorbent B prepared in Preparation Example 4, and adsorbent C prepared in Preparation Example 7, with a mass ratio of 5:4:1;

[0119] (3) Preparation of control stock solutions: Codeine, morphine, papaverine, thebaine, and narcotine stock solutions were prepared at concentrations of 50.0 mg / L, 50.0 mg / L, 10.0 mg / L, 10.0 mg / L, and 10.0 mg / L, respectively;

[0120] Prepare stock solution 1 (codeine 5000 ng / mL, morphine 5000 ng / mL, papaverine 1000 ng / mL, narcotine 1000 ng / mL, thebaine 1000 ng / mL) as follows: Accurately pipette 0.5 mL of stock solution into a 5 mL volumetric flask, dilute to the mark with solvent, mix, and label as stock solution 1.

[0121] Prepare L-1000% solution (codeine 1250 ng / mL, morphine 1250 ng / mL, papaverine 250 ng / mL, narcotine 250 ng / mL, thebaine 250 ng / mL) as follows: accurately pipette 5.0 mL of stock solution 1 into a 20 mL volumetric flask, dilute to the mark with solvent, mix, and label as L-1000% solution;

[0122] (4) Preparation of reference solution: Prepare reference solution (codeine 125 ng / mL, morphine 125 ng / mL, papaverine 25 ng / mL, narcotine 25 ng / mL, thebaine 25 ng / mL) as follows: accurately pipette 1.0 mL of L-1000% solution into a 10 mL volumetric flask, dilute to the mark with solvent, and mix thoroughly to prepare the reference solution.

[0123] (5) Sensitivity preparation: Prepare sensitivity solution (codeine 25 ng / mL, morphine 25 ng / mL, papaverine 5 ng / mL, narcotine 5 ng / mL, thebaine 5 ng / mL) as follows: accurately pipette 0.2 mL of L-1000% solution into a 10 mL volumetric flask, dilute to the mark with solvent and mix thoroughly to prepare the sensitivity solution;

[0124] (6) Detection: Determination of codeine, morphine, papaverine, narcotine, and thebaine in poppy oil using LC-MS / MS;

[0125] The chromatographic conditions for the detection are as follows: chromatographic column: Agilent ZORBAX SB-Aq 3.5μm 3.0×100mm, column temperature 40°C, injection volume 5μL, injection plate temperature 5°C, mobile phase: A: 5Mmol / L ammonium acetate + 0.1% formic acid (v / v); B: 0.1% formic acid acetonitrile solution (v / v), flow rate 0.3mL / min, needle wash solution MeOH:H2O=1:1 (v:v); mass spectrometry conditions: ion source ESI, curtain gas 35psi, temperature 600°C, ionization voltage 5500V, spray gas 50psi, auxiliary heating gas 50psi, scanning mode positive ion mode.

[0126] Example 2

[0127] This embodiment provides a method for determining the content of codeine, morphine, papaverine, narcotine, and thebaine in poppy oil, comprising the following steps:

[0128] (1) Preparation of isotope internal standard working solutions: Accurately pipette appropriate amounts of morphine-D3 isotope solution and codeine-D3 isotope solution into volumetric flasks, and dilute to the mark with water to prepare internal standard working solutions with a concentration of 5.0 μg / mL for both morphine-D3 and codeine-D3.

[0129] (2) Preparation of test solution: Add 1 g of adsorbent to 20 g of poppy oil sample, stir and adsorb for 20 min, filter, add the solid to the solvent, heat to 45 °C, stir and desorb for 40 min, filter, wash, combine the washing liquid and the filtrate, add 2 μg of isotope internal standard working solution, filter through a 0.22 μm filter membrane, and adjust the volume to prepare the test solution;

[0130] The solvent is a mixed solution of formic acid and acetonitrile with a volume ratio of 1:100;

[0131] The adsorbent includes adsorbent A prepared in Preparation Example 2, adsorbent B prepared in Preparation Example 5, and adsorbent C prepared in Preparation Example 8, with a mass ratio of 7:6:3;

[0132] (3) Preparation of control stock solutions: Codeine, morphine, papaverine, thebaine, and narcotine stock solutions were prepared at concentrations of 50.0 mg / L, 50.0 mg / L, 10.0 mg / L, 10.0 mg / L, and 10.0 mg / L, respectively;

[0133] Prepare stock solution 1 (codeine 5000 ng / mL, morphine 5000 ng / mL, papaverine 1000 ng / mL, narcotine 1000 ng / mL, thebaine 1000 ng / mL) as follows: Accurately pipette 0.5 mL of stock solution into a 5 mL volumetric flask, dilute to the mark with solvent, mix, and label as stock solution 1.

[0134] Prepare L-1000% solution (codeine 1250 ng / mL, morphine 1250 ng / mL, papaverine 250 ng / mL, narcotine 250 ng / mL, thebaine 250 ng / mL) as follows: accurately pipette 5.0 mL of stock solution 1 into a 20 mL volumetric flask, dilute to the mark with solvent, mix, and label as L-1000% solution;

[0135] (4) Preparation of reference solution: Prepare reference solution (codeine 125 ng / mL, morphine 125 ng / mL, papaverine 25 ng / mL, narcotine 25 ng / mL, thebaine 25 ng / mL) as follows: accurately pipette 1.0 mL of L-1000% solution into a 10 mL volumetric flask, dilute to the mark with solvent, and mix thoroughly to prepare the reference solution.

[0136] (5) Sensitivity preparation: Prepare sensitivity solution (codeine 25 ng / mL, morphine 25 ng / mL, papaverine 5 ng / mL, narcotine 5 ng / mL, thebaine 5 ng / mL) as follows: accurately pipette 0.2 mL of L-1000% solution into a 10 mL volumetric flask, dilute to the mark with solvent and mix thoroughly to prepare the sensitivity solution;

[0137] (6) Detection: Determination of codeine, morphine, papaverine, narcotine, and thebaine in poppy oil using LC-MS / MS;

[0138] The chromatographic conditions for the detection are as follows: chromatographic column: Agilent ZORBAX SB-Aq 3.5μm 3.0×100mm, column temperature 40°C, injection volume 5μL, injection plate temperature 5°C, mobile phase: A: 5Mmol / L ammonium acetate + 0.1% formic acid (v / v); B: 0.1% formic acid acetonitrile solution (v / v), flow rate 0.3mL / min, needle wash solution MeOH:H2O=1:1 (v:v); mass spectrometry conditions: ion source ESI, curtain gas 35psi, temperature 600°C, ionization voltage 5500V, spray gas 50psi, auxiliary heating gas 50psi, scanning mode positive ion mode.

[0139] Example 3

[0140] This embodiment provides a method for determining the content of codeine, morphine, papaverine, narcotine, and thebaine in poppy oil, comprising the following steps:

[0141] (1) Preparation of isotope internal standard working solutions: Accurately pipette appropriate amounts of morphine-D3 isotope solution and codeine-D3 isotope solution into volumetric flasks, and dilute to the mark with water to prepare internal standard working solutions with a concentration of 5.0 μg / mL for both morphine-D3 and codeine-D3.

[0142] (2) Preparation of test solution: Add 1 g of adsorbent to 20 g of poppy oil sample, stir and adsorb for 20 min, filter, add the solid to the solvent, heat to 42 °C, stir and desorb for 30 min, filter, wash, combine the washing liquid and the filtrate, add 2 μg of isotope internal standard working solution, filter through a 0.22 μm filter membrane, and adjust the volume to prepare the test solution;

[0143] The solvent is a mixed solution of formic acid and acetonitrile with a volume ratio of 0.7:100;

[0144] The adsorbent includes adsorbent A prepared in Preparation Example 3, adsorbent B prepared in Preparation Example 6, and adsorbent C prepared in Preparation Example 9, with a mass ratio of 6:5:2;

[0145] (3) Preparation of control stock solutions: Codeine, morphine, papaverine, thebaine, and narcotine stock solutions were prepared at concentrations of 50.0 mg / L, 50.0 mg / L, 10.0 mg / L, 10.0 mg / L, and 10.0 mg / L, respectively;

[0146] Prepare stock solution 1 (codeine 5000 ng / mL, morphine 5000 ng / mL, papaverine 1000 ng / mL, narcotine 1000 ng / mL, thebaine 1000 ng / mL) as follows: Accurately pipette 0.5 mL of stock solution into a 5 mL volumetric flask, dilute to the mark with solvent, mix, and label as stock solution 1.

[0147] Prepare L-1000% solution (codeine 1250 ng / mL, morphine 1250 ng / mL, papaverine 250 ng / mL, narcotine 250 ng / mL, thebaine 250 ng / mL) as follows: accurately pipette 5.0 mL of stock solution 1 into a 20 mL volumetric flask, dilute to the mark with solvent, mix, and label as L-1000% solution;

[0148] (4) Preparation of reference solution: Prepare reference solution (codeine 125 ng / mL, morphine 125 ng / mL, papaverine 25 ng / mL, narcotine 25 ng / mL, thebaine 25 ng / mL) as follows: accurately pipette 1.0 mL of L-1000% solution into a 10 mL volumetric flask, dilute to the mark with solvent, and mix thoroughly to prepare the reference solution.

[0149] (5) Sensitivity preparation: Prepare sensitivity solutions (codeine 25 ng / mL, morphine 25 ng / mL, papaverine 5 ng / mL, narcotine 5 ng / mL, thebaine 5 ng / mL) as follows: Accurately pipette 0.2 mL of L-1000% solution into a 10 mL volumetric flask, dilute to the mark with solvent, and mix thoroughly to prepare the sensitivity solution. The limit of detection (LOD) is also called sensitivity; the detection limit refers to the lowest amount of the analyte that can be detected in the sample. This is the limit of quantification (LOQ) preparation method; the quantification limit refers to the lowest amount of the analyte that can be quantitatively determined in the sample.

[0150] (6) Detection: Determination of codeine, morphine, papaverine, narcotine, and thebaine in poppy oil using LC-MS / MS;

[0151] The chromatographic conditions for the detection are as follows: chromatographic column: Agilent ZORBAX SB-Aq 3.5μm 3.0×100mm, column temperature 40°C, injection volume 5μL, injection plate temperature 5°C, mobile phase: A: 5Mmol / L ammonium acetate + 0.1% formic acid (v / v); B: 0.1% formic acid acetonitrile solution (v / v), flow rate 0.3mL / min, needle wash solution MeOH:H2O=1:1 (v:v); mass spectrometry conditions: ion source ESI, curtain gas 35psi, temperature 600°C, ionization voltage 5500V, spray gas 50psi, auxiliary heating gas 50psi, scanning mode positive ion mode.

[0152] Comparative Example 1-2

[0153] Compared with Example 3, the difference is that adsorbent A is prepared by Comparative Preparation Example 1-2.

[0154] Comparative Examples 3-5

[0155] Compared with Example 3, the difference is that adsorbent B is prepared by Comparative Preparation Examples 3-5.

[0156] Comparative Examples 6-7

[0157] Compared with Example 3, the difference is that adsorbent C is prepared by Comparative Preparation Examples 6-7.

[0158] Example 4

[0159] (1) Limit of detection (LOD):

[0160] The results are shown in Table 1.

[0161] Table 1

[0162] ;

[0163] As can be seen from the table above, the signal-to-noise ratio meets the requirements. The signal-to-noise ratio is used in analytical methods that display baseline noise. This ratio compares the signal from a known low-concentration sample with the signal from a blank sample to calculate the lowest concentration or amount of the substance that can be reliably detected. The detection limit is generally determined by the concentration or amount injected into the instrument at a signal-to-noise ratio of 3:1.

[0164] (2) Limit of quantification (LOQ):

[0165] The results are shown in Table 2. The limit of quantification is generally determined by the concentration corresponding to a signal-to-noise ratio of 10:1 or the amount injected into the instrument.

[0166] Table 2

[0167] ;

[0168] It can be seen from the above table that the signal-to-noise ratio of the quantification limit meets the requirements.

[0169] (3) Linear range: The linear range is within the range of LOQ to 200% limit concentration, and the regression coefficient (r) is ≥0.990, indicating good linearity.

[0170] The results are shown in Table 3.

[0171] Table 3

[0172] ;

[0173] (4) Accuracy

[0174] Three sample spike solutions were prepared at three accuracy limits of 50%, 100%, and 150%. The average recoveries for the corresponding concentrations ranged from 70.0% to 130.0%. The precision of the recoveries for the nine spiked sample solutions was no greater than 15.0%. The results are shown in Tables 4 to 8.

[0175] Table 4 Codeine

[0176] ;

[0177] Table 5 Morphine

[0178] ;

[0179] Table 6 Papaverine

[0180] ;

[0181] Table 7 Nascotine

[0182] ;

[0183] Table 8 Thebaine

[0184] ;

[0185] Test Example 1

[0186] The average recoveries and precisions of the spiked test results for codeine, morphine, papaverine, narcotine, and thebaine in the same batch of poppy oil samples (detected using the methods in Example 3 and Comparative Examples 1-7, respectively) are shown in Tables 9-13.

[0187] Table 9 Codeine

[0188] ;

[0189] Table 10 Morphine

[0190] ;

[0191] Table 11 Papaverine

[0192] ;

[0193] Table 12 Nascotine

[0194] ;

[0195] Table 13 Thebaine

[0196] ;

[0197] As can be seen from the above table, the average recoveries of codeine, morphine, papaverine, narcotine and thebaine determined by the methods of Examples 1-3 of the present invention are 93.60-98.96%, and the precisions are 1.2-3.3%.

[0198] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for determining the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil, characterized in that: The following steps are involved: (1) Preparation of isotope internal standard working solutions: Accurately pipette morphine-D3 isotope solution and codeine-D3 isotope solution into volumetric flasks, and adjust the volume to prepare the internal standard working solutions of morphine-D3 and codeine-D3 respectively; (2) Preparation of test solution: add adsorbent to poppy oil sample, stir for adsorption, filter, add solid to solvent, heat for desorption, filter, wash, combine washing solution and filtrate, add isotope internal standard working solution, filter, and adjust to volume to prepare test solution; (3) Preparation of control stock solutions: Prepare corresponding stock solutions using codeine, morphine, papaverine, thebaine, and narcotine standards; (4) Preparation of reference solution: dilute the reference stock solution to prepare the reference solution; (5) Detection: LC-MS / MS was used to determine the contents of codeine, morphine, papaverine, narcotine and thebaine in poppy oil.

2. The measuring method according to claim 1, wherein The adsorbent includes adsorbent A, adsorbent B and adsorbent C, with a mass ratio of 5-7:4-6:1-3; The preparation method of the adsorbent A is as follows: Morphine, thebaine, monomer, cross-linking agent, emulsifier and porogen are mixed and added to ethyl acetate, mixed evenly, added dropwise to water, emulsified, and under inert gas protection, an initiator is added, heated and stirred to react, the product is filtered, added to a Soxhlet extractor, extracted with a mixed solvent until no morphine and thebaine are detected, washed, and dried to prepare adsorbent A; The preparation method of the adsorbent B is as follows: S1. Preparation of a chiral amino acid-coupled MOF: L-aspartic acid and L-glutamic acid were added to water, followed by N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and activated by stirring. UiO-66-NH2 was added, the reaction was stirred, filtered, washed, and dried to obtain a chiral amino acid-coupled MOF. S2. Double bond grafting modification: Maleic acid was added to water, followed by N-hydroxysuccinimide and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and the mixture was stirred for activation. A chiral amino acid was added to couple the MOF, and the mixture was stirred for reaction. The mixture was filtered, washed, and dried to obtain the modified chiral amino acid-coupled MOF. S3. Copolymerization: The modified chiral amino acid-coupled MOF, methyl methacrylate, styrene, and N-isopropylacrylamide were mixed in ethyl acetate, uniformly mixed, and dropwise added to water for emulsification. Under inert gas protection, an initiator was added, and the reaction was heated with stirring. The product was filtered to produce adsorbent B. The preparation method of the adsorbent C is as follows: T1 Preparation of diazonium sulfanilate solution: sulfanilic acid and sodium nitrite were dissolved in lye, heated and mixed, cooled to room temperature, hydrochloric acid was added dropwise, and the reaction was stirred to obtain a diazonium sulfanilate solution; T2. Preparation of sulfonated / double-bond co-modified graphene oxide: Maleic acid was added to toluene, graphene oxide and p-toluenesulfonic acid were added, the mixture was heated to reflux with stirring, the reaction was centrifuged, washed, and dried, and the product was added to a solution of diazonium p-aminobenzenesulfonic acid, the reaction was stirred, centrifuged, washed, and dried to obtain sulfonated / double-bond co-modified graphene oxide; T3. Copolymerization: Sulfonated / double-bond co-modified graphene oxide, methyl methacrylate, styrene, and N-isopropylacrylamide were mixed in ethyl acetate, uniformly mixed, and dropwise added to water for emulsification. Under inert gas protection, an initiator was added, and the mixture was heated and stirred for reaction. The product was filtered to obtain adsorbent C.

3. The measuring method according to claim 2, wherein In the preparation process of adsorbent A, the monomers include methyl methacrylate, styrene, 1-vinyl-3-(aminopropyl) imidazole bis(trifluoromethanesulfonyl) imide salt, and N-isopropyl acrylamide, with a mass ratio of 3-5:4-6:1-2:5-7, the crosslinker is ethylene glycol dimethacrylate, the emulsifier is a mixture of Tween-80 and Span-80, with a mass ratio of 2-3:3-5, the porogen is hexadecyltrimethylammonium chloride or hexadecyltrimethylammonium bromide, and the initiator is The cross-linking agent is selected from at least one of azobisisobutyronitrile, sodium persulfate, potassium persulfate, and ammonium persulfate. The temperature of the heating and stirring reaction is 55-65° C. and the time is 4-6 hours. The mass ratio of morphine, thebaine, monomer, cross-linking agent, emulsifier, porogen and initiator is 0.5-1:0.5-1:8-12:0.2-0.4:0.5-1:0.1-0.2:0.01-0.

15. The mixed solvent is a mixed solvent of acetonitrile and dichloromethane, and the volume ratio is 1:1-3.

4. The measuring method according to claim 2, wherein The mass ratio of L-aspartic acid, L-glutamic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and UiO-66-NH2 in step S1 is 2-3:1-2:2-4:2-4:10; the mass ratio of maleic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, and chiral amino acid coupling MOF in step S2 is 2-4:1-2:1-2:10; the mass ratio of the modified chiral amino acid coupling MOF, methyl methacrylate, styrene, N-isopropylacrylamide, and initiator in step S3 is 5-7:4-6:3-5:6-8:0.1-0.2, the temperature of the heating and stirring reaction is 60-70°C, the time is 3-5h, and the initiator is selected from at least one of sodium persulfate, potassium persulfate, and ammonium persulfate.

5. The measuring method according to claim 2, wherein The mass ratio of p-aminobenzenesulfonic acid, sodium nitrite and hydrochloric acid in step T1 is 0.04-0.06:1:6-8, and the stirring reaction time is 3-5 hours; the mass ratio of maleic acid, graphene oxide, p-toluenesulfonic acid and p-aminobenzenesulfonic acid diazonium salt solution in step T2 is 2-3:8-10:0.5-1:15-20, the heating reflux stirring reaction time is 4-7 hours, and the stirring reaction time is 10-12 hours; the mass ratio of sulfonated / double bond co-modified graphene oxide, methyl methacrylate, styrene, N-isopropylacrylamide and initiator in step T3 is 4-6:3-5:3-4:5-7:0.1-0.2, and the heating stirring reaction time is 55-65°C for 4-6 hours.

6. The measuring method according to claim 1, wherein The chromatographic conditions for the detection are as follows: chromatographic column: Agilent ZORBAX SB-Aq 3.5μm 3.0×100mm, column temperature 35-45°C, injection plate temperature 3-7°C, mobile phase: A: 2-7Mmol / L ammonium acetate + 0.05-0.15% formic acid (v / v); B: 0.05-0.15% formic acid acetonitrile solution (v / v), flow rate 0.1-0.5mL / min, needle wash solution MeOH:H2O=1:1 (v:v).

7. The measuring method according to claim 1, wherein The mass spectrometry conditions for the detection are: ion source ESI, curtain gas 30-40 psi, temperature 550-650° C., ionization voltage 5000-6000 V, spray gas 45-55 psi, auxiliary heating gas 45-55 psi, and scanning mode positive ion mode.

8. The measuring method according to claim 1, wherein in, The linear range of codeine was 62.5 ng / mL-187.5 ng / mL, the linear range of morphine was 62.5 ng / mL-187.5 ng / mL, the linear range of papaverine was 12.5 ng / mL-37.5 ng / mL, the linear range of narcotine was 12.5 ng / mL-37.5 ng / mL, and the linear range of thebaine was 12.5 ng / mL-37.5 ng / mL.

9. The measuring method according to claim 1, wherein in, The regression coefficients for codeine were 0.995, morphine 0.994, papaverine 0.995, narcotine 0.998, and thebaine 0.

999.

10. The measuring method according to claim 1, wherein The mass ratio of the poppy oil, the adsorbent, and the isotope internal standard working solution is 15-20:1-2:0.000001-0.000002; the solvent is a mixed solution of formic acid and acetonitrile with a volume ratio of 0.5-1:100; the heating desorption temperature is 40-45° C., and the time is 20-40 minutes.