CN115477591A - Isotope drug labeled compound and preparation method and application thereof - Google Patents

Isotope drug labeled compound and preparation method and application thereof Download PDF

Info

Publication number
CN115477591A
CN115477591A CN202211318637.6A CN202211318637A CN115477591A CN 115477591 A CN115477591 A CN 115477591A CN 202211318637 A CN202211318637 A CN 202211318637A CN 115477591 A CN115477591 A CN 115477591A
Authority
CN
China
Prior art keywords
sample
detection
drug
labeled compound
detected
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202211318637.6A
Other languages
Chinese (zh)
Inventor
孙喆明
石寒天
林何
疏天民
洪利军
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Third Research Institute of the Ministry of Public Security
Original Assignee
Third Research Institute of the Ministry of Public Security
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Third Research Institute of the Ministry of Public Security filed Critical Third Research Institute of the Ministry of Public Security
Priority to CN202211318637.6A priority Critical patent/CN115477591A/en
Publication of CN115477591A publication Critical patent/CN115477591A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C225/00—Compounds containing amino groups and doubly—bound oxygen atoms bound to the same carbon skeleton, at least one of the doubly—bound oxygen atoms not being part of a —CHO group, e.g. amino ketones
    • C07C225/20—Compounds containing amino groups and doubly—bound oxygen atoms bound to the same carbon skeleton, at least one of the doubly—bound oxygen atoms not being part of a —CHO group, e.g. amino ketones having amino groups bound to carbon atoms of rings other than six-membered aromatic rings of the carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B59/00—Introduction of isotopes of elements into organic compounds ; Labelled organic compounds per se
    • C07B59/001—Acyclic or carbocyclic compounds
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C221/00—Preparation of compounds containing amino groups and doubly-bound oxygen atoms bound to the same carbon skeleton
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/28—Control of physical parameters of the fluid carrier
    • G01N30/34—Control of physical parameters of the fluid carrier of fluid composition, e.g. gradient
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G01N30/7233—Mass spectrometers interfaced to liquid or supercritical fluid chromatograph
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/05—Isotopically modified compounds, e.g. labelled
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/12—Systems containing only non-condensed rings with a six-membered ring
    • C07C2601/14—The ring being saturated
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N2030/042—Standards
    • G01N2030/045—Standards internal

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)

Abstract

The isotope drug labeling compound provided by the invention can be used for internal standard for measuring the content of drug fludrolone in detection materials such as sewage, hair and the like, and can be used for qualitatively detecting the fludrolone according to retention time and ion pair matching, and particularly, quantitatively detecting the corresponding compound according to the chromatographic peak area. The isotope drug labeled compound is used as an internal standard substance, can reduce the matrix effect of a detection material, and has good application prospect in aspects such as judicial identification and the like.

Description

Isotope drug labeled compound and preparation method and application thereof
Technical Field
The invention belongs to the technical field of preparation and application of standard substances of mental active substances, and particularly relates to an isotopic mental active substance labeled compound, and a preparation method and application thereof.
Background
Aiming at the detection of trace and trace substances in a detected material, an isotope dilution mass spectrometry method of a liquid chromatography-mass spectrometer combined with an isotope labeled standard substance internal standard is the most common and efficient detection means. The isotope labeled standard substance is used as an internal standard, and the isotope labeled internal standard and a compound to be detected have completely consistent chemical properties except molecular weight, so that the isotope dilution mass spectrometry can achieve high detection precision and sensitivity, and is an indispensable tool for detecting the chemicals in the sewage.
The detection of drugs and related metabolites in the sewage can comprehensively and intuitively evaluate the inundation situation of the toxicity situation of a city, and the detection is combined with the control of related data of toxicity enforcement work, so that the method is one of important means for urban overall toxicity situation evaluation and criminal investigation, and plays a key role in urban toxicity situation evaluation and key attack. The detection of the drugs and related metabolites in the hair can reflect the long-term exposure condition of the detected object to the drugs, and is an essential detection means for drug addiction judgment, community drug rehabilitation and the like.
At present, the method for detecting trace amount of flunomide in environmental or biological detection materials is generally a liquid phase mass spectrometry, and the content of the flunomide in the detection materials is determined by a standard curve method by selecting a compound with similar chemical properties to the flunomide as an internal standard. The internal standard of choice is typically the more common methoxamine, D4-ketamine or D3-diazepam.
Therefore, it is an urgent need to solve the problems of the prior art to provide an isotope drug labeling compound for improving the accuracy and sensitivity of drug detection in a test material.
Disclosure of Invention
Aiming at the problems of low drug detection precision and low sensitivity in the prior art, the invention provides an isotope drug labeling compound, which has the following structural formula:
Figure BDA0003910466430000011
wherein R is 1 、R 2 、R 3 At least one is D, and the drug contains fludrolone.
Preferably, the structural formula of the isotope drug labeling compound is:
Figure BDA0003910466430000021
the second aspect of the present invention provides a method for preparing the isotope drug labeled compound, wherein the method comprises the following steps:
(1) Synthesis of N-benzyl norflunomide ketone by reaction of norflunomide ketone and benzyl chloride
Figure BDA0003910466430000022
(2) Reacting N-benzyl norfluridone with deuterated iodomethane to synthesize N-benzyl-D3-fluridone:
Figure BDA0003910466430000023
(3) Reducing the N-benzyl-D3-fluoroamidone in the step (1) to D3-fluoroamidone under deuterium gas conditions:
Figure BDA0003910466430000024
the third aspect of the invention provides the application of the isotope drug labeled compound in detecting the content of the isotope drug flunomide in biological detection materials or sewage.
In the above application, the method for detecting the content of the flutriaxone in the biological sample or the sewage by using the isotope narcotic labeled compound comprises the following steps:
(1) Setting the detection conditions of liquid chromatography-mass spectrometry;
(2) Drawing a standard curve: adding different proportions of a standard solution of the fludrolone and the D3-fludrolone into a negative test material corresponding to a sample to be detected, performing LC-MS/MS detection after the same pretreatment step as the sample to be detected, and making a standard curve for the peak area ratio and the concentration of the fludrolone and the D3-fludrolone; wherein the internal standard is an isotopically drug-labeled compound of claim 1 or 2.
(3) Pretreatment and determination of a sample to be detected: adding the isotope drug labeled compound in claim 2 into a sample to be detected as an internal standard, performing LC-MS/MS detection after the same pretreatment steps as those used for drawing a standard curve to obtain quantitative parameters of the sample to be detected and the internal standard, and calculating by using a formula of the standard curve of the internal standard method to obtain the content of the sample to be detected.
Preferably, in step (1), the detection conditions of the liquid chromatography-mass spectrometry detection are as follows: the mobile phase is as follows: acetonitrile (0.01% formic acid), B: water (0.01% formic acid, 5% ammonium formate), gradient: 1min 80% by weight, A,4min 95% by weight, A,10min 95% by weight; a chromatographic column: poroshell120 PFP 3.0x100mm 1.9um; column temperature: 50 ℃; flow rate: 0.5mL/min; sample introduction amount: 5 mu L of the solution; an ion source: electrospray ion source, positive mode (ESI +); the spraying voltage is 3500V; ion source temperature: 340 ℃; collision gas: and (4) nitrogen.
Preferably, in the step (2), the mass concentration of the internal standard substance in the mixed standard solution is 0.1-99.9%.
Preferably, in the step (3), the mass concentration of the internal standard substance in the sample to be detected is 0.1-99.9%.
Preferably, in step (3), the formula of the internal standard method standard curve is as follows:
Figure BDA0003910466430000031
wherein X is the concentration of a sample to be detected, Y is the peak area obtained by liquid chromatography-mass spectrometry, b0 is the intercept of a standard curve, and b1 is the slope of the standard curve.
Preferably, the mass content of fludrolone in the biological sample or the sewage is 10 -7 ％～10％。
The three hydrogen atoms on the N-methyl group of the flunomide ketone molecule are all in equivalent positions for mass spectrum detection, namely the C-H bond or C-D bond on the methyl group is not involved in the fragmentation process of ions in mass spectrum, and the methyl structure is in a stable state. Therefore, the different substitution modes of the three atoms on the N-methyl do not influence the performance of the N-methyl in mass spectrum, and the corresponding mass spectrum parameters can be simply obtained by referring to D3-flunomide ketone molecules.
In another aspect, the present invention provides a method for detecting the content of drug fludrolone in biological detection materials or sewage, the method comprises using isotope drug labeled compound as internal standard to detect drug fludrolone, the structural formula of the compound is as follows:
Figure BDA0003910466430000032
preferably, the method for detecting the content of the flutriafol in the biological detection material or the sewage comprises the following steps:
(1) Setting detection conditions of liquid chromatography-mass spectrometry;
(2) Preparing a mixed standard solution containing a sample to be detected and an internal standard substance, performing LC-MS/MS detection, and making a standard curve for peak area ratio and concentration of the fludrolone and D3-fludrolone; wherein the internal standard is the isotopically drug-labeled compound of claim 1 or 2.
(3) Pre-treating and measuring a sample to be measured, adding the isotope drug labeled compound in claim 2 after pre-treating the sample to be measured as an internal standard, performing LC-MS/MS detection to obtain quantitative parameters of the sample to be measured and the internal standard, and calculating by using an internal standard method formula to obtain the content of the sample to be measured.
The isotope drug labeling compound D3-flunomide provided by the invention can be used as an internal standard for measuring the content of the isotope drug flunomide in sewage and biological detection materials, can reduce the matrix effect of the detection materials, and has good application prospects in aspects such as judicial identification and the like.
Drawings
FIG. 1 shows the high resolution mass spectrum of D3-fluoroamidone in example 1 of the present invention.
FIG. 2A Process for preparing D3-Fluoroaminone in example 1 of the present invention 1 H-NMR spectrum.
FIG. 2B preparation of D3-Fluoroaminone in example 1 of the present invention 13 C-NMR spectrum.
FIG. 3 is an extracted ion chromatogram of D3-fluoroamidone of example 1 of the present invention.
FIG. 4 is a chromatogram for detecting a fadrofreon sample in example 2 of the present invention.
FIG. 5 is a chromatogram for detecting a fadroflumetone sample in example 2 of the present invention.
FIG. 6 is a hair fluorenone calibration curve for example 2 of the present invention.
FIG. 7 is a chromatogram for detecting a fluoroamidone sample from wastewater in example 3 of the present invention.
FIG. 8 is a chromatogram for detecting a fluoroamidone sample from wastewater in example 3 of the present invention.
FIG. 9 shows a standard curve of the sewage flumethazone in example 3 of the present invention.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further explained below by combining the specific drawings.
In the description of the present invention, the "biological sample" refers to a human tissue sample. Such as hair, blood, urine, etc.
In the present description, the term "matrix effect" refers to the influence and interference of substances other than the analyte flunomide on the analysis process and the detection result
In the description of the invention, the mass concentration of the fludrolone in the biological detection material or the sewage is 10 -7 ％～10％。
The three hydrogen atoms on the N-methyl group of the flunominone molecule are all in equivalent positions for mass spectrum detection, namely the C-H bond or the C-D bond on the methyl group is not involved in the ion fragmentation process in mass spectrum, and the methyl structure is in a stable state.
Therefore, the different substitution modes of the three atoms on the N-methyl group do not influence the performance of the N-methyl group in the mass spectrum, and the corresponding mass spectrum parameters can be simply obtained by referring to the D3-flunomide molecule. In the present invention, the isotope drug labeled compound D3-flunomide is taken as an example for illustration, and the structural formula is as follows:
Figure BDA0003910466430000051
the invention is further illustrated by the following specific examples.
Example 1
A preparation method of D3-flunomide ketone.
To a solution of norflunomide (222mg, 1mmol) and triethylamine (0.4 mL) in dichloromethane (5 mL) at 0 deg.C was added dropwise a solution of benzyl chloride (139mg, 1.1mmol) in dichloromethane (5 mL), and after all the additions, the temperature was returned to room temperature and the reaction was allowed to proceed for 4 hours. Washing the organic phase with water, saturated sodium bicarbonate water solution and saturated salt solution, and evaporating the solvent from the organic phase to obtain oily matter, namely the intermediate N-benzyl norfluridone. N-benzyl norfluridone and cesium carbonate (1 mmol) were dissolved in DMF (10 mL), deuterated iodomethane (0.6 mmol) was added dropwise, stirred at room temperature for 2 hours, then heated to 50 ℃ and heated for 2 hours. After cooling, the solvent was removed by rotary evaporation, and 5mL of dichloromethane was added to dissolve the solution, and 5% mmol of Pd/C (10%) was added, and the solution was heated to reflux for 5 hours under reduction of 1atm of deuterium gas, and then cooled to 0 ℃ and water was added dropwise. The insoluble matter was removed by Celite filtration, and the Celite was washed with 10mL of methylene chloride, and the organic phases were combined, washed with a 5% aqueous solution of sodium hydroxide and dried over anhydrous magnesium sulfate. The drying agent was removed by filtration and the solvent removed by rotary evaporation. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 1) gave D3-fluoroaminoketone as a colorless liquid (50mg, 45%). 1H NMR (600MHz, methanol-D4) delta 7.81 (td, J =7.9,1.7Hz, 1H), 7.67 (dddd, J =8.6,7.1,5.3,1.6Hz, 1H), 7.49 (td, J =7.7,1.3Hz, 1H), 7.34 (ddd, J =11.8,8.3,1.2Hz, 1H), 3.32-3.28 (M, 1H), 2.57-2.49 (M, 2H), 2.16-2.08 (M, 1H), 1.95 (ddd, J =15.4,10.1,3.7Hz, 2H), 1.82-1.66 (M, 2H). 13C NMR (151MHz, methanol-D4) delta 205.24,205.23,162.05,160.40,133.46,133.40,130.28,130.27,125.79,125.76,117.75,117.67,117.00,116.85,69.36,38.44,38.43,34.30,28.47,21.33.HR-MS (ESI/TOF) M/z: calcd.for C13H14D3FNO [ M + H ] +225.1482; 225.1476. The spectrogram is shown in the attached figures 1,2A,2B and 3.
Example 2
And detecting the content of the flunomide in the hair by taking the D3-flunomide as an internal standard.
(1) Liquid chromatography-mass spectrometry detection conditions:
a) The instrument model is as follows: agilent 1290-6470QQQ;
b) And (3) chromatographic column: poroshell120 PFP 3.0x100mm 1.9um;
c) Column temperature: 50 ℃;
d) Mobile phase: acetonitrile (0.01% formic acid), B: water (0.01% formic acid, 5% ammonium formate), gradient: 1min 80% by weight, A,4min 95% by weight, A,10min 95% by weight;
e) Flow rate: 0.5mL/min;
f) Sample injection amount: 5 mu L of the solution;
g) An ion source: electrospray ion source, positive mode (ESI +);
h) The spraying voltage is 3500V;
i) Ion source temperature: 340 ℃;
j) Collision gas: nitrogen gas.
The ion pairs and corresponding conditions are shown in table 1:
TABLE 1
Figure BDA0003910466430000061
(2) Pretreatment and detection of samples
Cleaning hair sample with ultrapure water, detergent water and acetone, air drying, cutting into pieces, weighing 20.0mg, adding 1mL methanol (containing 1ng/mL D3-fluoroaminoketone), grinding, ultrasonic treating in a refrigerated ultrasonic instrument for 30min, centrifuging at 4000r for 5min, collecting supernatant 800 μ L, volatilizing in water bath air flow at 60 deg.C, redissolving with 80 μ L methanol, filtering with 0.22 μ L filter membrane, and subjecting 5 μ L to LC-MS/MS analysis to obtain the spectrum shown in figure 4 and 5. Wherein the quantitative ion pair is 191.0/222.0, and the peak area ratio of the flunomide to the internal standard D3-flunomide in two detections is 12162053/1203546 respectively.
(3) Drawing of standard curve
Cleaning blank negative hair samples with ultrapure water, detergent water and acetone, drying in the air, cutting into pieces, weighing 20.0mg of each part, adding 1mL of methanol (containing 1ng/mL of D3-fluoroamidone), adding a fluoroamidone standard reference substance, preparing into hair addition samples with the concentrations of 0.005, 0.01, 0.05, 0.1, 0.2, 0.5, 1.0, 2.0 and 10.0ng/mg, preparing 3 parts of each concentration in parallel, swirling for 3min, standing and soaking for 30min at room temperature, treating according to a sample pretreatment process, performing LC-MS/MS detection, and making standard curves for peak area ratios and concentrations of fluoroamidone and D3-fluoroamidone to obtain a graph shown in figure 6. The standard curve formula is Y =18.202398 xx-1.208354, wherein X is the concentration of the sample to be detected, Y is the peak area obtained by liquid chromatography-mass spectrometry detection, and b is 0 =1.208354 is the intercept of the standard curve, b 1 =18.202398 is the slope of the standard curve.
According to the quantitative relation between the peak area ratio and the concentration in the standard curve and the calculation formula:
Figure BDA0003910466430000071
and calculating the content of the fludrolone in the sample to be detected to be 0.49ng/mg.
Example 3
And detecting the content of the flunomide in the sewage by taking the D3-flunomide as an internal standard.
(1) Liquid chromatography-mass spectrometry detection conditions
Same as in example 1.
(2) Pretreatment and detection of samples
Filtering the sewage sample by filter paper, taking 100mL, adding 2mL of methanol (containing 10ng/mL D3-fluoroamidone), passing 50mL of each sample through an SPE column at the speed of 5mL/min, eluting with 5mL of methanol after the sample is finished, finally eluting with 5mL of 5% ammonia-acetonitrile solution, volatilizing the eluent under water bath air flow at 60 ℃, redissolving with 80 μ L of methanol, passing through a 0.22 μ L filter membrane, and performing LC-MS/MS analysis on 5 μ L of the eluent to obtain a spectrum shown in an attached figure 7 and an attached figure 8. Wherein the quantitative ion pair is 191.0/222.0, and the peak area ratio of the flunomide to the internal standard D3-flunomide in two detections is 43526/846298 respectively.
(3) Drawing of standard curve
Adding 1mL of methanol (containing 10ng/mL of D3-fluoroamidone) into 50mL of a negative sewage sample, adding a fluoroamidone standard reference substance, preparing 2 parts of sewage adding samples with the concentrations of 0.5, 1,2, 5, 10, 20 and 50ng/L in parallel at each concentration, treating the samples according to a sample pretreatment process, performing LC-MS/MS detection, and making a standard curve for the peak area ratio and the concentration of the fluoroamidone and the D3-fluoroamidone to obtain a graph shown in figure 6. The standard curve formula is Y =0.005675 xx +0.016637, wherein X is the concentration of the sample to be detected, Y is the peak area obtained by liquid chromatogram-mass spectrum detection, b 0 =0.016637 is the intercept of the standard curve, b 1 =0.005675 is the slope of the standard curve.
According to the quantitative relation between the peak area ratio and the concentration in the standard curve and the calculation formula:
Figure BDA0003910466430000072
and calculating the content of the flunomide in the sample to be detected to be 6ng/L according to the quantitative relation between the peak area ratio and the concentration in the standard curve.
When the isotope drug labeling compound provided by the invention is used for detecting a drug, namely, the fludrolone, in a detection material, a proper amount of the isotope drug labeling compound D3-fludrolone is added into the detection material, appropriate pretreatment is carried out according to the detection requirement, then liquid chromatography-mass spectrometry (LC-MS/MS) detection is carried out, the isotope drug labeling compound is used as an internal standard, and qualitative and quantitative detection of the to-be-detected substance is realized by comparing the peak area ratios of a target substance and the to-be-detected substance in a Multiple Reaction Monitoring (MRM) mode, so that the detected drug is the fludrolone, and the isotope drug can be detected, and has the advantages of strong specificity and high sensitivity.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.

Claims (10)

1. An isotopically labeled compound, wherein the compound has the following structural formula:
Figure FDA0003910466420000011
wherein R is 1 、R 2 、R 3 At least one is D, and the drug contains fludrolone.
2. The isotopically labeled compound of claim 1, wherein the isotopically labeled compound has the formula:
Figure FDA0003910466420000012
3. a method for preparing an isotopically labeled compound according to claim 1 or 2, comprising the steps of:
(1) Synthesis of N-benzyl norflunomide ketone by reaction of norflunomide ketone and benzyl chloride
Figure FDA0003910466420000013
(2) The N-benzyl norfluridone reacts with deuterated iodomethane to synthesize the N-benzyl-D3-fluridone:
Figure FDA0003910466420000014
(3) Reducing the N-benzyl-D3-fluoroaminoketone in the step (1) into D3-fluoroaminoketone under the deuterium gas condition:
Figure FDA0003910466420000015
4. use of the isotopically drug-labeled compound of claim 1 or 2 for detecting the content of a toxic fluoroamidone in a biological sample or in waste water.
5. The use according to claim 4, wherein the method for detecting the content of the flunomide in the biological sample or the sewage by using the isotope drug labeled compound according to claim 1 or 2 comprises the following steps:
(1) Setting the detection conditions of liquid chromatography-mass spectrometry;
(2) Drawing a standard curve: adding different proportions of a standard solution of flutolamine and D3-flutolamine into a negative detection material corresponding to a sample to be detected, performing LC-MS/MS detection after the same pretreatment step as the sample to be detected, and making a standard curve for the peak area ratio and the concentration of the flutolamine and the D3-flutolamine, wherein an internal standard substance is the isotope drug labeled compound in claim 1 or 2;
(3) Pretreatment and determination of a sample to be detected: adding the isotope drug labeled compound in claim 2 into a sample to be detected as an internal standard substance, performing LC-MS/MS detection after the same pretreatment steps as those used for drawing a standard curve to obtain quantitative parameters of the sample to be detected and the internal standard substance, and calculating by using a formula of the standard curve of the internal standard method to obtain the content of the sample to be detected.
6. The method according to claim 5, wherein in the step (1), the detection conditions of the liquid chromatography-mass spectrometry detection are as follows: mobile phase: acetonitrile (0.01% formic acid), B: water (0.01% formic acid, 5% ammonium formate), gradient: 1min 80% by weight, A,4min 95% by weight, A,10min 95% by weight; a chromatographic column: poroshell120 PFP 3.0x100mm 1.9um; column temperature: 50 ℃; flow rate: 0.5mL/min; sample introduction amount: 5 mu L of the solution; an ion source: electrospray ion source, positive mode (ESI +); the spraying voltage is 3500V; ion source temperature: 340 ℃; collision gas: nitrogen gas.
7. The method according to claim 5, wherein in the step (2), the mass concentration of the internal standard substance in the mixed standard solution is 0.1-99.9%; in the step (3), the mass concentration of the internal standard substance in the sample to be detected is 0.1-99.9%.
8. The method according to claim 5, wherein in the step (3), the formula of the standard curve by the internal standard method is as follows:
Figure FDA0003910466420000021
wherein X is the concentration of a sample to be detected, Y is the peak area obtained by liquid chromatography-mass spectrometry detection, b 0 Is the intercept of the standard curve, b 1 Is the slope of the standard curve.
9. The use according to claim 4, wherein the content of the fluoroamidone in the biological sample or wastewater is 10 -7 ％～10％。
10. A method for detecting the content of drug flunomide in biological detection materials or sewage comprises the step of detecting the drug flunomide by using an isotope drug labeled compound as an internal standard substance, wherein the structural formula of the compound is as follows:
Figure FDA0003910466420000031
CN202211318637.6A 2022-10-26 2022-10-26 Isotope drug labeled compound and preparation method and application thereof Pending CN115477591A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202211318637.6A CN115477591A (en) 2022-10-26 2022-10-26 Isotope drug labeled compound and preparation method and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202211318637.6A CN115477591A (en) 2022-10-26 2022-10-26 Isotope drug labeled compound and preparation method and application thereof

Publications (1)

Publication Number Publication Date
CN115477591A true CN115477591A (en) 2022-12-16

Family

ID=84395255

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202211318637.6A Pending CN115477591A (en) 2022-10-26 2022-10-26 Isotope drug labeled compound and preparation method and application thereof

Country Status (1)

Country Link
CN (1) CN115477591A (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101765582A (en) * 2007-04-26 2010-06-30 奥斯拜客斯制药有限公司 Deuterium-labelled ketamine
CN113698307A (en) * 2021-09-08 2021-11-26 上海义守生物科技有限公司 Isotope compound and preparation method and application thereof
CN113717066A (en) * 2021-09-08 2021-11-30 上海义守生物科技有限公司 Isotope compound and preparation method and application thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101765582A (en) * 2007-04-26 2010-06-30 奥斯拜客斯制药有限公司 Deuterium-labelled ketamine
CN113698307A (en) * 2021-09-08 2021-11-26 上海义守生物科技有限公司 Isotope compound and preparation method and application thereof
CN113717066A (en) * 2021-09-08 2021-11-30 上海义守生物科技有限公司 Isotope compound and preparation method and application thereof

Similar Documents

Publication Publication Date Title
CN113717066A (en) Isotope compound and preparation method and application thereof
Eaton et al. Kinetic isotope effect on proton tautomerism in tetraarylporphyrins
CN113321611A (en) Isotope mental active substance labeled compound and preparation method and application thereof
CN113698307A (en) Isotope compound and preparation method and application thereof
CN112305007B (en) Method for measuring deuterium isotope abundance of deuterium labeled compound by using nuclear magnetic hydrogen spectrum or deuterium spectrum
CN112608270B (en) Isotope compound and preparation method and application thereof
Zhao et al. Sensitive and accurate determination of neurotransmitters from in vivo rat brain microdialysate of Parkinson's disease using in situ ultrasound-assisted derivatization dispersive liquid–liquid microextraction by UHPLC-MS/MS
Zhang et al. A near-infrared BODIPY-based fluorescent probe for the detection of hydrogen sulfide in fetal bovine serum and living cells
Chen et al. A novel single-fluorophore-based ratiometric fluorescent probe for detection of formaldehyde in air
CN102980968A (en) Liquid chromatogram tandem mass spectrum measuring method for creatinine in urine
CN115784995A (en) Isotopic psychoactive substance labeled compound, preparation method and application thereof
Uzu et al. Use of 4-(N, N-dimethylaminosulphonyl)-7-fluoro-2, 1, 3-benzoxadiazole as a labelling reagent for peroxyoxalate chemiluminescence detection and its application to the determination of the β-blocker metoprolol in serum by high-performance liquid chromatography
Shima et al. Direct determination of glucuronide and sulfate of 4-hydroxy-3-methoxymethamphetamine, the main metabolite of MDMA, in human urine
CN115477591A (en) Isotope drug labeled compound and preparation method and application thereof
CN103389351B (en) Method for simultaneously detecting 9 acetylene-terminated hormones by virtue of liquid phase-tandem mass spectrometry
CN115047093B (en) Method for detecting dimethyl sulfate in anhydrous caffeine
Minami et al. Novel tryptamine-related substances, 5-sulphatoxydiacetyltryptamine, 5-hydroxydiacetyltryptamine, and reduced melatonin in human urine and the determination of those compounds, 6-sulphatoxymelatonin, and melatonin with fluorometric HPLC
CN115677444A (en) Isotope drug metabolite marker compound, preparation method and application thereof
CN111830175B (en) Method for detecting micro-molecular aldehyde in air
CN114478389A (en) Isotopic psychoactive substance labeled compound, preparation method, application and application thereof
Zhao-Fu et al. Flow injection-chemiluminescence method for determination of hydrocortisone in human serum by using trivalent silver complex
CN113325088A (en) Isotope mental active substance labeled compound and preparation method and application thereof
CN113325089A (en) Isotope mental active substance labeled compound and preparation method and application thereof
CN115894377A (en) Isotopic psychoactive substance labeled compound, preparation method and application thereof
CN110563609B (en) Preparation method and application of near-infrared fluorescent probe for detecting selenious acid roots

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination