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A kind of assay method of PEG content in biological sample

A method of determination, a technology of biological samples, applied in the field of determination of PEG content in biological samples, can solve the problems of not fundamentally solving the problems, the reduction of quantitative accuracy, and the increase of operation steps, so as to improve the specific identification ability, improve the sensitivity and The effect of accuracy

Active Publication Date: 2017-03-29
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, before quantification, the proportion of PEG with each degree of polymerization to the total amount must be determined before quantification can be completed, and a standard curve must be prepared separately for each component, and the operation steps will increase accordingly, resulting in a decrease in the accuracy of quantification
The above two methods both use the MRM mode, and still need to use Q1 to select the precursor ion, which has not fundamentally solved the problem, and there are still some shortcomings

Method used

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  • A kind of assay method of PEG content in biological sample
  • A kind of assay method of PEG content in biological sample
  • A kind of assay method of PEG content in biological sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] A method for determining PEG content in a biological sample, which is determined by liquid chromatography-time-of-flight mass spectrometry (LC-Q-Q-TOF), and the determination steps include:

[0039] A. Establish a standard curve for PEG determination;

[0040] B. Use liquid chromatography-time-of-flight mass spectrometry to measure the sample to be tested, and calculate the concentration of PEG in the sample to be tested by the standard curve obtained in step A;

[0041] 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, and the settings of the mass spectrometry part of steps A and B are: the voltage in the first mass analyzer Q1 does not select precursor ions, charged particles After all passes, enter the second mass analyzer Q2; set the collision energy in the second mass analyzer Q2 to break the charged particles into fragment ions; selec...

Embodiment 2

[0055] On the basis of Example 1, the determination of PEG 4000 in plasma was specifically selected for quantification.

[0056] The preparations before the measurement are:

[0057] Collection of Rat Plasma Samples

[0058] ① Select one male rat with a body weight of 200 g ± 10 g, free to eat and drink;

[0059] ② Weigh PEG 4000 and dilute to 1.0 mg / mL with normal saline;

[0060] ③The administration process is as follows: tail vein injection administration, the administration dose is 0.6 mL of 1.0 mg / mL PEG 4000 normal saline solution per mouse. Rats were treated before administration (0 h) and after administration at 0.083 h, 0.25 h, 0.5 h, 0.75 h, 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 10 h, 12 h , 24 h, take 0.5 mL of blood through the orbit of the rat, put the whole blood in a pre-cooled heparinized EP tube, centrifuge at 4°C (13300 rpm, 5 min), separate and transfer all the plasma to another EP tube Stored in a -20°C freezer for testing.

[0061] Determination of PEG 4000 C...

Embodiment 3

[0091] The determination of the content of PEG 10000 in the plasma sample is the same as in Example 2 except that the following parameters are different from Example 2.

[0092] In the preparatory work before the determination, the rats were injected with PEG 10000 saline solution, and the standard product selected when making the standard solution was PEG 10000. Among the determination parameters: in the mass spectrometry condition, the unclustering voltage is 100 V, and the collision voltage is 30 eV; the chromatographic elution program is shown in Table 6. See Tables 7 and 8 for standard curves and accuracy.

[0093]

[0094]

[0095]

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Abstract

The invention relates to a method for determining PEG content in a biological sample. The method includes: liquid chromatography-time of flight mass spectroscopy is used for determining the PEG content, a standard curve is produced, and the standard curve is used to calculate the PEG concentration in the to-be-determined sample. The mass spectrum condition of the method is based on the triple tandem mass spectrum technology. The mass spectrum part is characterized in that parent ions are not selected in a first mass analyzer Q1, and all charged particles enter a second mass analyzer Q2 after passing; collision energy is set in the second mass analyzer Q2 to smash the charged particles into fragment ions; stable feature fragment ions are selected in a third mass analyzer which is a time of flight mass analyzer to quantify PEG. The method has the advantages that the method aims at the non-uniqueness of the PEG molecular weight in the biological sample, and the method is simple and practical, accurate and reliable in result, high in sensitivity and good in reproducibility.

Description

technical field [0001] The invention belongs to the technical field of drug analysis, in particular to a method for measuring PEG content in biological samples. A method based on high performance liquid chromatography-time-of-flight mass spectrometry (LC-Q-Q-TOF) combined technology to determine the content of PEG with different degrees of polymerization in samples. Background technique [0002] Polyethylene glycol (PEG) is a pH-neutral, non-toxic, hydrophilic polymer with unique physical and chemical properties and good biocompatibility. It is also one of the few FDA-approved synthetic polymers that can be injected into the body one. When PEG is coupled to proteins, peptides, small molecule organic drugs or nanoparticle shells, it can reduce the immune clearance of pharmaceutical preparations and rapid elimination by the kidneys, prolong the drug's circulation time in vivo, and reduce the toxicity of the drug. As a new type of pharmaceutical excipient, the quality control...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 顾景凯周晓彤程龙妹尹磊杨艳
Owner JILIN UNIV
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