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In-vivo biomarker for detecting 5F-AMB and application thereof

A 5F-AMB and biomarker technology, which is applied in the field of synthetic cannabinoid 5F-AMB biomarkers, can solve the problems of detection method lag and achieve accurate and reliable results, high sensitivity and high precision

Pending Publication Date: 2020-12-11
ZHEJIANG POLICE COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In contrast to the grim situation of synthetic cannabinoids, there are currently few reports on the pharmacology and toxicology of synthetic cannabinoids, and the detection methods are lagging behind

Method used

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  • In-vivo biomarker for detecting 5F-AMB and application thereof
  • In-vivo biomarker for detecting 5F-AMB and application thereof
  • In-vivo biomarker for detecting 5F-AMB and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: The method of high-performance liquid phase tandem electrostatic orbital ion trap high-resolution mass spectrometry combined with liver microsome incubation experiment in vitro to obtain 5F-AMB metabolites in vitro:

[0036] (1) Take RLM (rat liver microsomes) and thaw at -20°C for 10 minutes, then thaw at 4°C for use;

[0037] (2) Accurately weigh 1mg of 5F-AMB, use DMSO as solvent, configure 1mg / ml 5F-AMB standard solution; take 20.4724mg NADP + (Oxidized coenzyme II sodium salt) in 1ml of PBS to get 0.026mmol / ml NADP + stock solution; get 20.0706mg G-6-P (glucose disodium 6-phosphate) 1ml in the PBS, obtain 0.066mmol / ml G-6-P stock solution; the G-6-PDH (glucose-6- Phosphate dehydrogenase) was dissolved in 10ml of PBS to obtain 100U / ml G-6-PDH stock solution; weigh 0.02852g of MgCl 2 Dissolve in 30ml of PBS to get 10mM MgCl 2 stock solution;

[0038] (3) In vitro incubation

[0039] 3.1 Take a 2ml centrifuge tube and add 10μl NADP + stock solution, 1...

Embodiment 2

[0046] Embodiment 2: Analysis of 5F-AMB metabolites in vitro

[0047] Enter the exact mass number of the parent drug in the total ion chromatogram, and extract the chromatographic peak; compare the response value of the chromatographic peak of the standard substance, the target substance without adding the standard, and the sample after incubation for 60 minutes, and obtain the change amount of the parent drug after incubation and metabolism, see The chromatogram of Fig. 15F-AMB, wherein (a) standard solution, (b) the sample incubated 60min, (c) blank solution.

[0048] figure 2 is the secondary mass spectrum of 5F-AMB;

[0049] image 3 is the chromatogram of liver microsome metabolites in vitro;

[0050] Figure 4 It is the secondary mass spectrogram of liver microsome in vitro metabolites and the deduced fragmentation mode;

[0051] Figure 5 is a schematic diagram of the in vitro metabolic pathway of liver microsomes;

[0052] Figure 6 is the curve of the 5F-AMB ...

Embodiment 3

[0069] Example 3: Validation of biomarkers by in vivo experiments in rats

[0070] Use polyoxyethylated castor oil and absolute ethanol at a ratio of 1:1 to prepare a 5 mg / ml 5F-AMB solution and dilute it to 1.25 mg / ml with normal saline for use. There were 2 male SD rats, and the rats were forbidden to eat 12 hours before the administration, but they were allowed to drink water freely; the weight of the rats was about 200g.

[0071] 0.8ml sample was injected through the tail vein, and the administration dose was 5mg / kg. After the tail vein injection, the rat urine was collected for 0h-4h, 4h-8h, 8h-24h, 24h-48h, 48h-72h. Stored at -80°C until analytical experiments, at which point the rats were sacrificed. Rats injected with blank solvent were used as control analysis.

[0072] Take a certain volume of urine and add 3 times the volume of acetonitrile, vortex for not less than 30s, centrifuge at a speed greater than 12000rpm for a certain period of time, take the supernatant...

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Abstract

The invention discloses an in-vivo biomarker for detecting 5F-AMB and application thereof. The in-vivo biomarker of synthetic cannabinoid 5F-AMB is detected as shown in Table 1. The in-vivo biomarkerfor detecting 5F-AMB and the application thereof overcome the defects of high in-vivo metabolism speed of the synthetic cannabinoid 5F-AMB and generally lower parent drug content in a biological detection material, and the invention provides a method for detecting by using an in-vivo metabolism marker, which has the advantages of high accuracy, high sensitivity and an accurate and reliable result.The biomass of the detection sample required by the invention is less and is only 100 microliters.

Description

technical field [0001] The present invention relates to the confirmation of a synthetic cannabinoid 5F-AMB biomarker, especially by using high-efficiency liquid phase series electrostatic orbital ion trap high-resolution mass spectrometry detection combined with liver microsome in vitro incubation experiment, through screening 5F-AMB in vitro metabolites, and analyzing 5F - AMB biomarkers, and validated by in vivo experiments in rats. Background technique [0002] Synthetic cannabinoids are a class of new psychoactive substances whose mechanism of action on the human body is similar to that of the natural cannabinoid Tetrahydrocannabinol (THC). Compared with natural cannabinoids, synthetic cannabinoids are more potent and harmful in the human body. Structurally speaking, many synthetic cannabinoids have nothing to do with classical cannabinoids; according to structure, synthetic cannabinoids can be roughly divided into seven categories, naphthoyl indole (JWH-018), naphthale...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/72G01N30/06G01N30/34G01N30/88G01N30/86
CPCG01N30/02G01N30/72G01N30/06G01N30/34G01N30/88G01N30/8634G01N2030/884G01N2030/8813
Inventor 柯星吕云平范一雷何丹丹周善慧
Owner ZHEJIANG POLICE COLLEGE
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