Method for detecting plasma protein binding rate of meropenem or imipenem by combining liquid chromatography-mass spectrometry technology with ultrafiltration technology

A technology of imipenem and meropenem, which can be used in measurement devices, instruments, scientific instruments, etc., can solve problems such as drug instability and inaccurate measurement results

Pending Publication Date: 2021-04-20
PEKING UNIV THIRD HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing methods for detecting the free concentration of meropenem, the determination results are inaccurate because the problem of drug instability is not solved

Method used

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  • Method for detecting plasma protein binding rate of meropenem or imipenem by combining liquid chromatography-mass spectrometry technology with ultrafiltration technology
  • Method for detecting plasma protein binding rate of meropenem or imipenem by combining liquid chromatography-mass spectrometry technology with ultrafiltration technology
  • Method for detecting plasma protein binding rate of meropenem or imipenem by combining liquid chromatography-mass spectrometry technology with ultrafiltration technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products. Example 1 Determination of plasma protein binding rate of meropenem or imipenem by liquid chromatography-mass spectrometry combined with ultrafiltration

[0045] 1 material

[0046] 1.1 Instrument

[0047] High performance liquid chromatography (DGU LC-30AD, Shimadzu Corporation); quadrupole / linear ion trap mass spectrometer (ABSciex 4000 + Mass spectrometry detector, American AB SCIEX company), equipped with electrospray ionization source (ESI); MS 3 digital vortex mixer (Germany IKA company); BT25S electronic balance (Germany Sartorius company); 3-18K centrifuge (US Sigma company); Cascada-I pure water machine (USA Pall company), HWT-10...

Embodiment 2

[0107] The optimization of embodiment 2 centrifugal force and centrifugal time

[0108] Add 200 μL and 400 μL plasma samples to the ultrafiltration tubes respectively, and centrifuge at 20°C for 20 minutes under the centrifugal force of 6000g, 8000g and 10000g for each volume of double samples (number 200-1, 200-2 and 400-1, 400-2 respectively). Or 30min, investigate the volume that obtains ultrafiltrate.The results are shown in Table 10:

[0109] The volume of ultrafiltrate under different centrifugal conditions of table 10

[0110]

[0111]

[0112] According to the standards that the ultrafiltration membrane is not damaged and the ratio of the ultrafiltrate to the total volume of plasma before ultrafiltration is 0.3-0.6, preferably 8000g is used as the centrifugal force, and the ultrafiltration time is in the range of 20-30min.

Embodiment 3

[0113] Embodiment 3 chromatographic column and the selection of mobile phase ratio

[0114] In this embodiment, the selection of the chromatographic column and the proportion of the mobile phase is carried out with the peak time and peak shape as the selection criteria.

[0115] Mobile phase: A: Acetonitrile (containing 0.1% formic acid); B: 5mM ammonium formate solution (containing 0.1% formic acid, 5% acetonitrile)

[0116] ①Symmetry C18 column (2.1×50mm, 3.5μm)

[0117] Mobile phase ratio: 90%B

[0118] Peak time: IMP: 0.38min; MER: 0.48min. IMP and MER are not retained on this column ( Figure 4 ).

[0119] ②Atlantis T3 column (2.1×50mm, 3μm)

[0120] Mobile phase ratio: 95%B

[0121] Peak time: IMP: 0.61min; MER: 2.42min. IMP is not retained on this column ( Figure 5 ).

[0122] ③HILIC Silica column (2.1mm×50mm, 3μm)

[0123] Mobile phase ratio: 15% B

[0124] Peak time: IMP: 3.68min; MER: 4.64min. Poor peak shape and poor sensitivity ( Figure 6 ).

[0125...

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Abstract

According to a method for detecting the plasma protein binding rate of meropenem or imipenem by combining a liquid chromatography-mass spectrometry technology with an ultrafiltration technology, the liquid chromatography-tandem mass spectrometry detection technology is combined with the ultrafiltration technology, and the total concentration of meropenem or imipenem in plasma and the free concentration of drugs in ultrafiltrate are respectively detected. And an MOPs stabilizer is added during sample treatment, so that the problem of poor drug stability is solved, and the determination result is more accurate. The method can be used for rapidly detecting the protein binding rate of the drug at high throughput, and is particularly suitable for unstable drug molecules.

Description

technical field [0001] The invention relates to the field of drug analysis, in particular to a method for determining the plasma protein binding rate of meropenem or imipenem by liquid chromatography-mass spectrometry combined with ultrafiltration. Background technique [0002] Meropenem (MEM) and imipenem (IMP) are carbapenem antibiotics, and their chemical structures are unstable. In the existing methods for detecting the free concentration of meropenem, the problem of drug instability is not solved, resulting in inaccurate determination results. Contents of the invention [0003] The purpose of the present invention is to provide a method for determining the plasma protein binding rate of meropenem or imipenem by liquid chromatography-mass spectrometry combined with ultrafiltration. [0004] In order to achieve the purpose of the present invention, the present invention provides a method for determining the plasma protein binding rate of meropenem or imipenem by liquid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/06G01N30/88
Inventor 王晨张现化杨平翟所迪许明哲赵荣生杨丽张斗胜崇小萌王立新姚尚辰王欣
Owner PEKING UNIV THIRD HOSPITAL
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