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Analysis method for detecting drug distribution and application thereof

An analysis method and drug technology, applied in the direction of analyzing materials, material analysis by electromagnetic means, measuring devices, etc., can solve the problems of difficult to truly reflect the distribution of drugs, unable to detect the distribution of drugs, etc., and achieve the effect of accurate drug distribution information.

Active Publication Date: 2017-11-24
宁波瑞瑧生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current commonly used drug distribution analysis methods can only detect the distribution of drugs at the tissue scale, but cannot detect the distribution of drugs in cells and subcellular structures, and it is difficult to truly reflect the distribution of drugs in the subject

Method used

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  • Analysis method for detecting drug distribution and application thereof
  • Analysis method for detecting drug distribution and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] The present embodiment provides an analytical method for detecting drug distribution, which includes:

[0051] Step a. Construct an in vitro myocardial tissue model: take the sterilized decellularized mouse myocardial tissue and place it in a 24-well plate whose inner wall is pre-coated with fibronectin; put the medium containing cardiomyocytes at a concentration of 50,000 cells / mL Inject the suspension into a 24-well plate; place the 24-well plate in an incubator for 4 hours, continue to add medium, and co-culture cardiomyocytes with decellularized mouse myocardial tissue for 7 days to obtain an in vitro myocardial tissue model.

[0052] Step b. Administration: the isotope 13 The C-labeled drug to be tested was added to the culture medium of the myocardial tissue model in vitro, and cultured for 48 hours under the same conditions.

[0053] Step c. Preparation and detection of samples to be tested: use 2.5% glutaraldehyde solution to chemically fix the in vitro myocard...

Embodiment 2

[0055] The present embodiment provides an analytical method for detecting drug distribution, which includes:

[0056] Step a. Constructing an in vitro stem cell co-culture model: take the sterilized decellularized porcine small intestinal mucosa tissue and place it in a 24-well plate whose inner wall is pre-coated with fibronectin; Inject the culture medium suspension into the 24-well plate; place the 24-well plate in the incubator for 4 hours, continue to add the medium, co-culture the mesenchymal stem cells with the decellularized porcine small intestinal mucosa tissue for 7 days, and obtain the in vitro Stem cell co-culture model.

[0057] Step b. Administration: the isotope 15 The N-labeled drug to be tested was added to the culture medium of the in vitro stem cell co-culture model, and cultured for 72 hours under the same conditions.

[0058] Step c. Preparation of the sample to be tested: After the in vitro stem cell co-culture model obtained in step b is subjected to ...

experiment example 1

[0061] Distribution detection of amiodarone in macrophages:

[0062] 1. Experimental method

[0063] 1. Construction of in vitro macrophage co-culture model: Decellularized natural soft tissue and macrophages (NR8383) were co-cultured in a culture dish.

[0064] 2. Administration: Amiodarone solution with a concentration of 1.56 μg / mL was added to the macrophage culture medium, and in 5% CO 2 and 37°C for 72 hours.

[0065] 3. Sample processing: use the chemical fixation method to process the co-culture model of macrophages after administration, that is, a solution prepared with 0.5% tannic acid and 2.5% glutaraldehyde (adjusted with 0.08m sodium cacodylate) pH to 7) to treat the macrophage co-culture model, followed by embedding with epoxy resin. After the resin was cured, cut into 500nm thin slices with a microscope microtome and dried on the surface of the silicon wafer.

[0066] 4. Electron microscope imaging: use a scanning electron microscope to detect the dried flak...

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Abstract

The present invention provides an analysis method for detecting drug distribution and application thereof, and belongs to the field of drug analysis and detection. The analysis method is as follows: using a fixing treatment method to treat a biological tissue administrated with a to-be-tested drug, and slicing the biological tissue to prepare a sample to be tested; and subsequently analyzing the sample to be tested by using a secondary ion mass spectrometer. Through the analysis method, the distribution of the to-be-tested drug in various tissues, cells and subcellular structures can be known, drug molecules can be detected in cell and subcellular scales, and accumulation of the to-be-tested drug in a target site can be known. The analysis method can be widely used in the study of detection of transport, absorption and metabolism of the drug, or in the study of detection of drug efficacy and drug screening.

Description

technical field [0001] The invention relates to the field of drug analysis and detection, in particular to an analysis method for detecting drug distribution and its application. Background technique [0002] Secondary ion mass spectrometer (SIMS) is a method to obtain material surface information by analyzing the secondary ions generated by sputtering after the primary ions are incident on the target surface by mass spectrometry. Secondary ion mass spectrometry can analyze all elements including hydrogen, and can give isotopic information, analyze compound components and molecular structures. Secondary ion mass spectrometry has high sensitivity, which can reach the order of ppm or even ppb, and can also perform micro-region composition imaging and depth profile analysis. [0003] The current commonly used drug distribution analysis methods can only detect the distribution of drugs at the tissue scale, but cannot detect the distribution of drugs in cells and subcellular str...

Claims

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Application Information

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IPC IPC(8): G01N27/62
CPCG01N27/62
Inventor 蒋海波陈琦
Owner 宁波瑞瑧生物科技有限公司
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