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Quantitative assay method of PEG-modified medicine in biological sample

A measurement method and technology of biological samples, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve problems such as non-unique values, and achieve the effects of strong specificity, wide application range, and high sensitivity

Active Publication Date: 2015-10-21
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] and PEG is a series of polymer mixtures composed of ethylene glycol as the basic unit, its molecular weight is not the only value, but the molecular weight of a certain degree of polymerization The average value is the center of normal distribution, for example: PEG 4000 is actually a mixture of PEG molecules with various molecular weights that are normally distributed around the molecular weight of 4000; PEG600, PEG4000, PEG6000, and PEG10000 are all mixtures of PEG molecules with various molecular weights; Therefore, this brings challenges to LC-MS / MS quantitative analysis based on the target molecular weight of the analyte

Method used

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  • Quantitative assay method of PEG-modified medicine in biological sample
  • Quantitative assay method of PEG-modified medicine in biological sample
  • Quantitative assay method of PEG-modified medicine in biological sample

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0034] A method for quantitative determination of PEGylated drugs in biological samples, which is determined by liquid chromatography-time-of-flight mass spectrometry, and the determination steps include:

[0035] A. Establish a standard curve for free drug determination and a standard curve for PEGylated drug determination,

[0036] B. Use liquid chromatography-time-of-flight mass spectrometry to measure the sample to be tested, and calculate the concentration of free drug and the concentration of PEGylated drug in the sample to be tested by the standard curve obtained in step A;

[0037] The chromatographic conditions and mass spectrometry conditions of steps A and B are the same, wherein the mass spectrometry conditions are based on triple tandem mass spectrometry technology, and the setting of the mass spectrometry part of steps A and B is: the voltage in the first mass analyzer Q1 is set to RF (radio frequency voltage) only, no parent ion is selected, all charged particle...

Embodiment 2

[0050] On the basis of Example 1, the PEGylated doxorubicin in plasma was specifically determined.

[0051] A quantitative assay method for PEGylated doxorubicin in a biological sample,

[0052] PEGylated doxorubicin with a drug loading equivalent to 1 mg of doxorubicin was dissolved in 1 mL of normal saline and administered to rats through the tail vein. 2, 3, 4, 6, 12, 24, 48, 72h. Determine the content of free doxorubicin and PEGylated doxorubicin in plasma after rat tail vein administration of PEGylated doxorubicin, the plasma drug concentration time curve is shown in figure 2 .

[0053] Measured by liquid chromatography-time-of-flight mass spectrometry, the determination steps include:

[0054] A. preparation of doxorubicin concentration determination standard curve and PEGylated doxorubicin concentration determination standard curve;

[0055] 1) Dilute doxorubicin and PEGylated doxorubicin stock solution to 0.05, 0.1, 0.3, 1.0, 3.0, 10.0, 30.0 μg / mL with acetonitr...

Embodiment 3

[0071] On the basis of Example 1, PEGylated gemcitabine in plasma was specifically determined.

[0072] A quantitative assay method for PEGylated gemcitabine in a biological sample,

[0073] PEGylated gemcitabine with a drug loading equivalent to 4 mg of gemcitabine was dissolved in 1 mL of normal saline and administered to rat tail veins. The blood collection time points were as follows: 0, 0.083h, 0.25h, 0.75h, 1h, 1.5h, 2h, 4h , 6 h, 9 h, 24 h, 36 h, take 0.5 mL of blood from the orbital vein of the rat, put the plasma in a pre-cooled heparinized EP tube, centrifuge at 4°C (15000 rpm, 5 min), separate the plasma, and take all Supernatant, the supernatant was stored in a -20°C refrigerator for testing. Determination of the content of free gemcitabine and PEGylated gemcitabine in the plasma after rat tail vein administration of PEGylated gemcitabine, the plasma drug concentration time curve is shown in Figure 4 .

[0074] Measured by liquid chromatography-time-of-flight...

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Abstract

The invention provides a quantitative assay method of PEG-modified medicine in a biological sample. Assay is conducted through liquid chromatogram-flight time mass spectrum. The method includes the steps that firstly, a standard curve is made; secondly, sample introduction assay is conducted on the sample to be tested, and the concentrate of the PEG-modified medicine and free medicine in the sample to be tested are worked out. The mass spectrum condition is based on a triple tandem mass spectrum technology, and the mass spectrum is set in the mode that parent ions are not selected in a first mass analyzer Q1, and charge particles enter a second mass analyzer Q2 after passing through the first mass analyzer Q1 totally; energy collision is set in the second mass analyzer Q2, and the charge particles are smashed into fragment ions; stable PEG-modified medicine feature fragment ions and free medicine feature fragment ions are selected in a third quality analyzer ion trap or flight time mass analyzer to quantitate the PEG-modified medicine and the free medicine. According to the method, for the nonuniqueness of PEG-modified medicine molecular weight in the biological sample, the quantitative assay method is established, and analysis and quantitation can be conducted on the PEG-modified medicine and the free medicine in the biological sample at the same time.

Description

technical field [0001] The invention belongs to the technical field of drug analysis and research, and specifically relates to a quantitative determination method for PEGylated drugs in biological samples. Background technique [0002] Polyethylene glycol (PEG) is an excellent water-soluble polymer compound with a neutral pH and non-toxicity. It is one of the few synthetic polymers approved by the FDA that can be injected into the body. Small molecule drugs, especially antineoplastic drugs, generally have defects such as poor water solubility, short half-life, poor targeting of biological tissue distribution, and high toxicity, which greatly limit their clinical applications. PEG is hydrophilic and has no immunogenicity. After PEG modification of drugs, the water solubility of drug molecules can be significantly improved, and the bioavailability of insoluble drugs can be improved. At the same time, due to the steric hindrance effect of PEG, the drug can be slowly released...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 顾景凯尹磊史美云孟祥骏宿崇彭雯雯陆欢林文孩杨艳景遐斌
Owner JILIN UNIV
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