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Method for Quantitative Analysis of Phosphane Isomers Using Liquid Chromatography-Ion Mobility Differential Mass Spectrometry

An ion mobility, quantitative analysis technology, applied in the field of biochemistry and pharmaceutical analytical chemistry, can solve the problems of inability to meet pharmacokinetics, poor analytical sensitivity, long analysis time, etc., to achieve high practicability and reliability, high sensitivity , The effect of strong data reproducibility

Active Publication Date: 2020-12-11
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In the prior art, only high performance liquid chromatography is used to separate and analyze the four isomers of Forzaine, but this method takes a long time to analyze, has poor specificity and poor analytical sensitivity, and cannot meet the requirements of pharmacokinetics. research requirements

Method used

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  • Method for Quantitative Analysis of Phosphane Isomers Using Liquid Chromatography-Ion Mobility Differential Mass Spectrometry
  • Method for Quantitative Analysis of Phosphane Isomers Using Liquid Chromatography-Ion Mobility Differential Mass Spectrometry
  • Method for Quantitative Analysis of Phosphane Isomers Using Liquid Chromatography-Ion Mobility Differential Mass Spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Quantitative analysis of four isomers of fudaine

[0038] This example illustrates the separation and quantification of four isomers of forsaine by high performance liquid chromatography-ion mobility differential mass spectrometry.

[0039] Solution preparation: Weigh 10 mg of Forsaine mixture powder, dissolve in 10 mL of N, N-dimethylformamide, prepare a solution with a concentration of 1 mg / mL, and use a diluent (methanol: acetonitrile: water = 25:25:50) Diluted to a solution of 800ng / mL, take 10-20 μL of the above solution and inject it into a high-performance liquid chromatography-ion mobility differential mass spectrometry tandem device to obtain a mass chromatogram ( figure 1 ). The four isomers of forsaine are completely separated by chromatography and ion differential two-dimensional separation, so they can be quantified simultaneously.

[0040] High performance liquid chromatography conditions:

[0041] Column: Waters UPLC BEH C 18 Column, spec...

Embodiment 2

[0052] Example 2: Methodological Validation

[0053] This example illustrates the methodological validation of the method for the quantitative analysis of the four isomers of Dacein in plasma.

[0054] Plasma sample preparation method:

[0055] Take 50 μL of plasma containing forsaine, add 200 μL of precipitant (methanol: acetonitrile = 50:50), centrifuge for 5 minutes, take 100 μL of supernatant, add 100 μL of water, mix, take 10-20 μL for injection.

[0056] Adopt the analysis method provided by the invention, use external standard method to carry out quantitative analysis. The method was validated for specificity, standard curve, lower limit of quantitation, precision and accuracy, matrix effects, and recovery.

[0057] A. Specificity: Individual plasma samples from 6 different volunteers were used to measure using the analysis method provided by the present invention. In the obtained mass chromatogram, it can be judged that there is no interference of endogenous substanc...

Embodiment 3

[0068] Embodiment 3: pharmacokinetic research

[0069] This example illustrates the study on the pharmacokinetics of four isomers after intravenous injection of forsaine injection in tumor patients.

[0070] Patients with advanced esophageal cancer were intravenously injected with 7 mg of forzaine for 1 hour, and 24 hours after administration, they were irradiated with 670nm laser. , 48h, 96h, 144h, 192h and 288h), blood samples (3 mL) were collected, which were centrifuged at 3000 g for 10 minutes at 4° C. to obtain plasma. Using the plasma pretreatment method provided by the present invention and the quantitative analysis method using high performance liquid chromatography-ion mobility differential mass spectrometry tandem device to analyze the pharmacokinetic characteristics of the four isomers of Forzaine in human plasma, to Study the clinical pharmacology of this innovative drug. The results of pharmacokinetic studies in image 3 shown in . Depend on image 3 As can ...

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Abstract

The invention provides a method for quantitatively analyzing four isomers FD-1, FD-2, FD-3 and FD-4 of photocyanine by a liquid chromatography-ion mobility differential mass spectrometry coupling technology. The method comprises the following steps: 1), preparing a photocyanine sample solution; 2), injecting the sample solution into a high performance liquid chromatography-ion mobility differential mass spectrometry serial device to obtain a mass chromatogram; 3), according to the mass chromatogram, quantitatively analyzing the four isomers FD-1, FD-2, FD-3 and FD-4 of the photocyanine by an external standard method, wherein in the high performance liquid chromatography, gradient elution is performed by using a mixed mobile phase consisting of a mobile phase A and a mobile phase B; the mobile phase A is a weakly alkali aqueous mobile phase having the pH value range of 9.5-11.5; the flow phase B is a mixture of methanol and acetonitrile.

Description

technical field [0001] The invention relates to the fields of biochemistry and pharmaceutical analytical chemistry, and more specifically relates to a method for quantitatively analyzing isomers of sarcine by using liquid chromatography-ion mobility differential mass spectrometry. Background technique [0002] Photodynamic therapy (PDT), as a new means of cancer treatment, has recently received extensive attention. The application of photodynamic therapy is based on three key factors: a photosensitizer, a specific wavelength of laser light, and oxygen. [0003] Forsaine is an amphiphilic disulfonic acid diphthalimide methyl phthalocyanine zinc dipotassium salt, composed of four isomers FD-1, FD-2, FD-3 and FD-4 composition. [0004] [0005] As a new type of anti-cancer phthalocyanine photosensitizer, the China State Food and Drug Administration (CFDA) approved the clinical trial of forsaine in June 2008, and is currently undergoing phase II clinical trials. According ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/72
CPCG01N30/02G01N30/72
Inventor 郑昕崔馨戈江骥
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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