Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for measuring CA4P and metabolites thereof

A technology for metabolites and samples, applied in the field of determination of combustin and its metabolites, can solve problems such as difficult separation, inconsistent mass spectrometry scanning modes, and inability to detect compounds to be tested, and achieve good reproducibility.

Active Publication Date: 2020-01-10
FUDAN UNIV SHANGHAI CANCER CENT
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the detection of Combustin and its metabolites in the body, due to the very similar chromatographic retention behavior of different compounds, it is difficult to separate, and the mass spectrometry scanning mode is inconsistent. It is necessary to switch between positive and negative ion scanning modes, so it is impossible to use a single mass spectrometry scanning mode to achieve simultaneous detection. Detect all test compounds
Therefore, so far, there is no report on the simultaneous determination of CA4P, CA4 and CA4G in human or animal plasma by quantitative analysis.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for measuring CA4P and metabolites thereof
  • Method for measuring CA4P and metabolites thereof
  • Method for measuring CA4P and metabolites thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Mass spectrometry ionization form of CA4P, CA4, CA4G and internal standard (deuterated CA4) under different ionization modes of embodiment 1

[0045] experimental method:

[0046] 1. CA4P standard, CA4 standard, CA4G standard and deuterated CA4 internal standard are equipped with aqueous solutions respectively, with a concentration of 1 μg / mL, mixed with the liquid phase mobile phase through the T-shaped tee by needle pump injection, and enter mass spectrometer.

[0047] 2. Liquid mobile phase: solvent A: H 2 O (containing 0.1% formic acid, 20 mM ammonium acetate); solvent B: methanol.

[0048]3. Liquid-mass spectrometry analysis conditions: chromatographic column: Waters SymmetryShield RP8 column (3.5μm, 50×2.1mm); column temperature: 40°C; elution condition: isocratic elution with 50% solvent B; flow rate: 0.30 mL / min; ESI(+) or ESI(-) ionization mode is used for mass spectrometry detection.

[0049] 4. Needle pump speed: 10μL / min.

[0050] Results: When using ES...

Embodiment 2

[0055] Example 2 Establishment of CA4P, CA4 and CA4G Quantitative Analysis Method in Human Plasma

[0056] With CA4P, CA4 and CA4G as the compounds to be tested and deuterated CA4 as the internal standard, a quantitative analysis method for CA4P, CA4 and CA4G in human plasma was established, and it was applied to human plasma after intravenous infusion of combustine preparations. Quantitative determination of the concentration of the test compound.

[0057] experimental method:

[0058] 1. Human plasma sample pretreatment:

[0059] To 50 μL human plasma sample, add 200 μL CH containing internal standard (40.0 ng / mL concentration) 3 CN-water solution (90:10, v / v) was used to precipitate proteins. After shaking for 5 min and centrifugation (4°C, 14,000 rpm, 10 min), the supernatant was taken for LC-MS / MS analysis, with an injection volume of 2 μL.

[0060] 2. Liquid-mass spectrometry analysis conditions: (the percentage content is volume percentage)

[0061] Chromatographic...

Embodiment 3

[0067] Example 3 Methodological verification of quantitative analysis of CA4P, CA4 and CA4G together

[0068] experimental method:

[0069] 1. Preparation of standard curve mixed working solution:

[0070] Precisely draw appropriate amount of CA4P, CA4 and CA4G standard stock solution (concentration is 1mg / mL), and use MeOH-H 2 O solution (50:50, v / v) was diluted to prepare standard working solutions with concentrations of 500 μg / mL (CA4P) and 100 μg / mL (CA4, CA4G). continue with MeOH-H 2 O solution (50:50, v / v) was diluted to make a mixed standard curve with concentrations ranging from 500 to 100,000 ng / mL (CA4P), 50.0 to 20,000 ng / mL (CA4) and 20.0 to 20,000 ng / mL (CA4G). Working solution, used to prepare standard curve plasma samples.

[0071] 2. Preparation of mixed quality control working solution

[0072] Precisely draw appropriate amount of CA4P, CA4 and CA4G standard stock solution (concentration is 1mg / mL), and use MeOH-H 2 O solution (50:50, v / v) was diluted to...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention provides a method for measuring CA4P and metabolites thereof. The method comprises a step of carrying out liquid chromatography-mass spectrometry on a to-be-detected sample, wherein a mobile phase containing ammonium acetate and formic acid is used in a high performance liquid chromatograph; and during mass spectrometry, an MRM mode is used for measuring the CA4P, a [M+H]<+> peak ofCA4 and a [M+NH4]<+> peak of CA4G in an ESI(+) ionization mode. According to the method for measuring the CA4P and the metabolites thereof, simultaneous detection of the CA4P, the CA4 and the CA4G ina single mass spectrum scanning mode becomes possible, an analysis process does not need to perform switching of positive and negative ion scanning modes of the mass spectrum, and the method the characteristics of specificity, sensitivity, simplicity, convenience, good reproducibility and the like.

Description

technical field [0001] The invention relates to the field of drug analysis, in particular to a method for measuring combustin and its metabolites. Background technique [0002] Combustine (CA4P) is an anti-angiogenesis anti-tumor drug developed by OXIGENE, USA, and is a water-soluble phosphate prodrug of CA4. CA4 is a cis-polyhydroxystilbene natural product isolated from the bark of the African shrub Combretum Caffrum. It is similar in structure to colchicine and can competitively bind to the colchicine binding site near β-microtubules , but the affinity is much higher than that of colchicine. However, the poor water solubility of CA4 affects its druggability. CA4P has greatly improved the water solubility of the drug due to the introduction of water-soluble phosphate groups. It does not combine with microtubules and has no anti-tumor activity, but it can be converted into its activity under the action of plasma non-specific and endogenous phosphatase Metabolite CA4, resu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/34G01N30/72
Inventor 孙艳吴奇珍王谦王漪璇黄京秋方雅玲吴维怡
Owner FUDAN UNIV SHANGHAI CANCER CENT