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Liposome drug in-vivo biological analysis method

A technology of bioanalysis and liposome, which is applied in the field of medicine, can solve the problems of not effectively reflecting the bioavailability of drugs, and achieve the effect of precise release degree

Active Publication Date: 2022-04-12
南京美新诺医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the complex interaction between drug release in liposomes and tissue and / or cell uptake of drugs and / or liposomes, only simple determination of Total drug concentration may not effectively reflect drug bioavailability in the intended target organ (i.e., site of action)

Method used

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  • Liposome drug in-vivo biological analysis method
  • Liposome drug in-vivo biological analysis method
  • Liposome drug in-vivo biological analysis method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Embodiment 1: the extraction process of total doxorubicin (low concentration quantitative range)

[0037] 1. After all the samples to be tested are completely melted, mix for 10-30 seconds.

[0038] 2. Pipette 50 μL of the sample to be tested into a 1.5 mL centrifuge tube, add 50 μL of IS solution with a concentration of 100 ng / mL, mix well, and obtain a sample to be tested containing an internal standard; pipette 50 μL of a blank sample (that is, 50 μL of blank plasma) to 1.5 Add 50 μL of IS solution with a concentration of 100 ng / mL to a mL centrifuge tube, mix well to obtain a blank internal standard control; pipette 50 μL of a blank sample (i.e. 50 μL of blank plasma) into a 1.5 mL centrifuge tube, add 50 μL of methanol:water (1 : 1), mix well, obtain blank control.

[0039]3. Add 800 μL of chloroform to the test sample containing internal standard, blank internal standard control, and blank control respectively, vortex for 3 minutes, and centrifuge at 10,000×g, 4°...

Embodiment 2

[0043] Embodiment 2: the extraction process of total doxorubicin (high concentration quantitative range)

[0044] 1. After all the samples to be tested are completely melted, mix for 10-30 seconds.

[0045] 2. Pipette 50 μL of the sample to be tested into a 1.5 mL centrifuge tube, add 50 μL of IS solution with a concentration of 500 ng / mL, mix well, and obtain a sample to be tested containing an internal standard; pipette 50 μL of a blank sample (that is, 50 μL of blank plasma) to 1.5 Add 50 μL of IS solution with a concentration of 500 ng / mL into a mL centrifuge tube, mix well to obtain a blank internal standard control; pipette 50 μL of a blank sample (i.e. 50 μL of blank plasma) into a 1.5 mL centrifuge tube, add 50 μL of methanol:water (1 : 1), mix well, obtain blank control.

[0046] 3. Add 800 μL of chloroform to the test sample containing internal standard, blank internal standard control, and blank control respectively, vortex for 3 minutes, and centrifuge at 10,000×g...

Embodiment 3

[0050] Embodiment 3: the extraction process of the doxorubicin of liposome encapsulation (low concentration quantitative range)

[0051] 1. After all the samples to be tested are completely melted, mix for 10-30 seconds.

[0052] 2. Add 100 μL of water to 50 μL of the sample to be tested and mix well to obtain a mixed sample.

[0053] 3. Sequentially add 1 mL of methanol and water to activate the SPE solid phase extraction plate (Strata C18-E).

[0054] 4. Transfer the above-mentioned mixed sample and 150 μL of water to the activated solid phase extraction plate in sequence, and collect the effluent.

[0055] 5. Add 50 μL of IS solution with a concentration of 100 ng / mL to the effluent, mix well, and obtain the test sample containing internal standard. Replace 50 μL of the blank sample (i.e. 50 μL of blank plasma) with the 50 μL of the sample to be tested in step 2 of this example and proceed to step 5 to obtain a blank internal standard control; replace the step of this exa...

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Abstract

The invention provides an in-vivo biological analysis method for a liposome drug. The total content of the liposome drug is detected by adopting a liquid-liquid extraction method and LC-MS / MS (Liquid Chromatography-Mass Spectrometry / Mass Spectrometry); a solid-phase extraction method, a liquid-liquid extraction method and LC-MS / MS are adopted to detect the content of the liposome wrapped medicine; and the content of the free liposome drug is obtained by subtracting the content of the liposome-coated drug from the total liposome drug content. According to the invention, after the liposome drug enters the body, the in-vivo free drug and the liposome wrapping part are effectively measured, and reference is provided for the in-vivo pharmacokinetic behavior of the liposome and the in-vivo release degree of the preparation. Compared with a traditional method for measuring the sum of the in-vivo lipidosome and the free medicine through protein precipitation, the method has the advantage that the release degree of the in-vivo lipidosome can be reflected more accurately.

Description

technical field [0001] The invention belongs to the field of medicine, in particular to a bioanalysis method for liposome medicine in vivo. Background technique [0002] Liposomes are vesicles composed of a single bilayer (unilamellar) and / or a series of concentric bilayers (multilamellar) separated by aqueous compartments formed by amphiphilic phospholipids, the closed center being aqueous Compartment. [0003] In liposomal drugs, the drug is usually encapsulated inside the liposome, generally water-soluble drugs are encapsulated in the aqueous compartment (1 or more), lipophilic drugs are encapsulated in the lipid bilayer Medium (1 or more). [0004] Liposomal drugs differ from emulsions, microemulsions, and drug-lipid mixtures in that the drug release behavior from liposomes, as well as other properties such as liposomal clearance and circulation half-life, may vary depending on polyethylene glycol and / or cholesterol or other The addition of possible liposome additives...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/02G01N30/72G01N1/34G01N1/40
CPCY02A50/30
Inventor 高峥贞张玲玲卢金莲
Owner 南京美新诺医药科技有限公司
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