CN114441696A

CN114441696A - Method for detecting eribulin mesylate key intermediate

Info

Publication number: CN114441696A
Application number: CN202111580570.9A
Authority: CN
Inventors: 田龙
Original assignee: Nanjing Gearpharma Technology Co ltd
Current assignee: Nanjing Gearpharma Technology Co ltd
Priority date: 2021-12-22
Filing date: 2021-12-22
Publication date: 2022-05-06

Abstract

The invention relates to a method for detecting an eribulin mesylate key intermediate. Provides a simple, convenient and sensitive liquid chromatography detection method for detecting the purity of the key intermediate of eribulin mesylate and LC-MS. The method adopts the tail-end ultraviolet absorption wavelength and adopts high performance liquid chromatography for detection, has high sensitivity, good separation degree and simple and convenient operation, is suitable for large-scale production central control analysis, and can be used as an LS-MS qualitative identification method.

Description

Method for detecting eribulin mesylate key intermediate

Technical Field

The invention relates to the field of drug detection, and particularly relates to a method for detecting an eribulin mesylate key intermediate. Belongs to the field of medicine technology.

Background

Eribulin mesylate (Eribulin) is a derivative of macrolide compound extracted from marine organism sponge, and is a substance with chemical activity. The medicinal action mechanism is that the medicinal composition directly combines with tubulin to inhibit mitosis, inhibits the growth of microtubules and inhibits the growth of cancer cells to play a therapeutic role. The clinical medicinal component of the medicine is eribulin mesylate, the trade name of which is Halaven, which is developed by Japanese Eisai company and is firstly marketed in the United states at 11/15/2010, and Halaven mesylate is approved by the United states Food and Drug Administration (FDA) to treat metastatic breast cancer patients who have received chemotherapy for advanced diseases at least twice, and the medicine is subsequently marketed in the countries of Singapore, European Union, Switzerland, Japan, China and the like.

The key intermediate of eribulin mesylate related to the patent has small polarity and weak ultraviolet absorption, and because the finished product eribulin mesylate is extremely valuable, the quality of the key intermediate of eribulin mesylate needs to be strictly controlled in order to improve and control the quality of the finished product eribulin mesylate.

The molecular structure of the key intermediate of eribulin mesylate is as shown in formula I:

at present, reports about key intermediate physique control methods of eribulin mesylate are not discovered at all. The method can be used as LC-MS detection method for identifying the product.

Disclosure of Invention

The invention aims to provide a simple and sensitive liquid chromatography detection method for detecting the purity of an eribulin mesylate key intermediate and LC-MS.

The technical scheme of the invention is as follows:

a method for detecting the purity and LC-MS of an eribulin mesylate key intermediate is characterized by comprising the following steps: the method comprises the steps of taking a butylsilane bonded silica gel filler (C4) as a chromatographic column, taking water-formic acid-acetonitrile as a mobile phase, and adopting gradient elution.

In the chromatographic conditions, 0.1-0.2% formic acid water solution is used as a mobile phase A, and 0.1-0.2% formic acid acetonitrile solution is used as a mobile phase B. The gradient elution procedure was as follows:

in the chromatographic conditions of the invention, the mobile phase A is 0.1% formic acid aqueous solution; the mobile phase B is 0.1 percent formic acid acetonitrile solution; the detection wavelength was 200nm and the column temperature was 25 ℃.

The invention further provides a purity and LC-MS detection method of the key intermediate of eribulin mesylate, which adopts a reversed-phase high performance liquid chromatography method and comprises the following steps:

1. preparing a test solution: taking a proper amount of the key intermediate of eribulin mesylate, adding methanol, dissolving to a constant volume, and preparing into a solution with a certain concentration;

2. and (3) chromatographic detection: using a butylsilane bonded silica gel filler (C4) as a chromatographic column, using a 0.1% formic acid aqueous solution as a mobile phase A, and using a 0.1% formic acid acetonitrile solution as a mobile phase B; the detection wavelength was 200nm and the column temperature was 25 ℃. The amount of sample was 5. mu.L.

Elution was performed using the following gradient elution procedure:

the product has very low polarity, very weak ultraviolet absorption and no obvious characteristic absorption, adopts the tail end ultraviolet absorption wavelength, is detected by a high performance liquid chromatography, has high sensitivity, good separation degree and simple and convenient operation, is suitable for large-scale production central control analysis, and can be used as an LS-MS qualitative identification method.

The LC-MS spectra of eribulin mesylate key intermediate are shown in figure 1-1, figure 1-2 and figure 1-3.

Drawings

FIGS. 1-1, 1-2 and 1-3 are LC-MS spectra of eribulin mesylate key intermediate

FIG. 2 is a liquid chromatogram of the key intermediate of eribulin mesylate in example 1

FIG. 3 is a liquid chromatogram of key intermediate of eribulin mesylate in example 2

DETAILED DESCRIPTION OF EMBODIMENT (S) OF INVENTION

The present invention will be described in further detail below with reference to examples and the accompanying drawings.

Example 1

In this example, C4(250 × 4.6,5 μm) was used as a chromatographic column, and methanol was used as a solvent, and the specific detection steps were as follows:

sample preparation: taking a proper amount of the key intermediate of eribulin mesylate, adding a solvent, ultrasonically dissolving and diluting to prepare a solution containing 1mg per 1mL, and shaking up.

Chromatographic conditions are as follows: using C4(250 x 4.6,5 μm) as chromatographic column, 0.1% formic acid water as mobile phase A, and 0.1% formic acid acetonitrile as mobile phase B; the detection wavelength is 200nm, the column temperature is 25 ℃, and the flow rate is 1.0 mL/min. The amount of sample was 5. mu.L.

Elution was performed using the following gradient elution procedure:

the liquid chromatogram of the key intermediate of eribulin mesylate is shown in FIG. 2.

Example 2

Chromatographic conditions are as follows: using C4(250 x 4.6,5 μm) as chromatographic column, 0.15% formic acid water as mobile phase A, and 0.15% formic acid acetonitrile as mobile phase B; the detection wavelength is 200nm, the column temperature is 25 ℃, and the flow rate is 1.0 mL/min. The amount of sample was 5. mu.L.

Elution was performed using the following gradient elution procedure:

the liquid chromatogram of the key intermediate of eribulin mesylate is shown in FIG. 3.

Claims

1. A method for detecting the purity and LC-MS of an eribulin mesylate key intermediate is characterized by comprising the following steps: the method comprises the steps of taking a butylsilane bonded silica gel filler (C4) as a chromatographic column, taking water-formic acid-acetonitrile as a mobile phase, and adopting gradient elution.

2. The detection method according to claim 1, wherein the chromatographic conditions include a mobile phase A comprising a 0.1% to 0.2% formic acid aqueous solution and a mobile phase B comprising a 0.1% to 0.2% formic acid acetonitrile solution.

3. The detection method according to claim 1, wherein the gradient elution procedure is as follows:

4. the detection method according to any one of claims 1 to 3, wherein the chromatographic conditions are such that the mobile phase A is a 0.1% aqueous formic acid solution.

5. The detection method according to any one of claims 1 to 3, wherein the chromatographic conditions are performed with a mobile phase B of 0.1% formic acid in acetonitrile.

6. The detection method according to any one of claims 1 to 3, wherein the chromatographic conditions are such that the detection wavelength is 200nm and the column temperature is 25 ℃.

7. A method for detecting the purity and LC-MS of an eribulin mesylate key intermediate by adopting a reversed-phase high performance liquid chromatography method comprises the following steps:

(1) preparing a test solution: taking a proper amount of the eribulin mesylate key intermediate, adding methanol, performing ultrasonic treatment, and dissolving;

(2) and (3) chromatographic detection: using C4 as chromatographic column, 0.1% formic acid water solution as mobile phase A, and 0.1% formic acid acetonitrile solution as mobile phase B; elution was performed using the following gradient elution procedure:

wherein the detection wavelength is 200nm, the column temperature is 25 ℃, and the flow rate is 1.0 mL/min; the amount of sample was 5. mu.L.