CN110508135B

CN110508135B - Capillary electrochromatography method for separating pantoprazole racemate

Info

Publication number: CN110508135B
Application number: CN201910641450.1A
Authority: CN
Inventors: 关瑾; 范实同; 石爽; 阎峰
Original assignee: Shenyang University of Chemical Technology
Current assignee: Shenyang University of Chemical Technology
Priority date: 2019-07-16
Filing date: 2019-07-16
Publication date: 2021-12-21
Anticipated expiration: 2039-07-16
Also published as: CN110508135A

Abstract

The invention provides a capillary electrochromatography method for separating pantoprazole racemate, and relates to a chiral separation method of drugs, which takes 6-p-toluenesulfonyl-beta-cyclodextrin as a chiral selector and bonds the chiral selector on the inner wall of a capillary by a chemical bonding method to prepare a chiral capillary electrochromatography column. Separation of pantoprazole enantiomers is achieved by capillary electrophoresis. Capillary electrochromatography separation conditions: the buffer salt is 50 mmol/Ltris-H₃PO₄pH 5.0; the separation voltage is 15 kV; the ultraviolet detection wavelength is 280 nm. The method is used for detecting the optical purity of the pantoprazole chiral medicine, and provides technical support for research and development of the chiral medicine.

Description

Capillary electrochromatography method for separating pantoprazole racemate

Technical Field

The invention relates to a chiral separation method of a medicament, in particular to a method for separating pantoprazole racemate.

Background

Pantoprazole is an anti-ulcer medicine and is mainly used for treating gastric acid secretion-related diseases such as duodenal ulcer, gastric ulcer, reflux esophagitis and the like. Sulfur atoms in the molecular structure of pantoprazole are chiral centers, two enantiomers of R and S exist, the activity of the S-isomer is stronger than that of the R configuration, and the research and development of single enantiomer of pantoprazole become hot spots. Capillary electrochromatography is a high-efficiency micro-separation chromatographic technique developed in recent decades, is a capillary electrophoresis mode for realizing separation based on chromatographic retention and electrodynamics principles, has the high efficiency of capillary electrophoresis and the high selectivity of liquid chromatography, and has wide application prospect in separation and analysis of enantiomers.

Capillary stationary phases are the heart of the capillary electrochromatography technique. Capillary electrochromatography columns can be divided into packed columns, monolithic columns and open columns, depending on the form of the stationary phase present. The open tubular column is used for bonding or coating the stationary phase on the inner wall of the capillary for electrochromatography separation. The invention is used for preparing a capillary electrochromatography open tubular column and is used for separating pantoprazole racemate.

Disclosure of Invention

The invention aims to provide a capillary electrochromatography method for separating pantoprazole racemate, which adopts chiral capillary electrochromatography to separate and analyze the pantoprazole racemate and provides technical support for research and development of pantoprazole single enantiomer medicaments.

The purpose of the invention is realized by the following technical scheme:

a capillary electrochromatography method for separating pantoprazole racemate is characterized in that 6-tosyl-beta-cyclodextrin is used as a chiral selector and is bonded to the inner wall of a capillary by a chemical bonding method to prepare a chiral capillary electrochromatography column. Separation of pantoprazole enantiomers is achieved by capillary electrophoresis.

The specific separation steps are as follows:

a, a column manufacturing mode:

(1) intercept an uncoated Quartz chromatography column (inner diameter 75)

) The column length is 53 cm, the column is burned at 8 cm, and then the column is wiped by acetone until the column is completely transparent, and the column is a detection window, and the effective length is 45 cm. Washing with pure methanol for 10 min; washing with distilled water for 30 min; washing with 1M NaOH for 1 h; washing with distilled water for 30 min; washing with absolute ethyl alcohol for 1 h; washing with 1% 3-aminopropyl-3-methoxysilane solution for 1 hr, sealing the column overnight, and oven drying in an oven at 100 deg.C for 24 hr.

(2) And washing the dried capillary tube with 1% 6-p-toluenesulfonyl-beta-CD-acetonitrile solution for 3 h, and standing for 1 h. Then washing with 50% anhydrous ethanol for 10min, and washing with 100% anhydrous ethanol for 30 min.

The flushing flow rate is 800 mu L/h

b. The electrochromatography separation conditions are as follows:

the background electrolyte was 50 mM tris-H₃PO₄Solution, pH 5.0; the separation voltage is 15 kV; ultraviolet detection wavelength: 280 nm. Before each analysis, 0.1M NaOH, redistilled water and running buffer solution are used respectivelyWashing for 10 min; and washing the sample injection room with running buffer solution for 5 min, and carrying out next sample injection.

The capillary electrochromatography method for separating pantoprazole racemate comprises the following capillary electrochromatography separation conditions: the buffer salt is 50 mmol/Ltris-H₃PO₄pH 5.0; the separation voltage is 15 kV; the ultraviolet detection wavelength is 280 nm.

The invention has the advantages and effects that:

the invention provides a capillary electrochromatography for separating pantoprazole racemate, which takes 6-tosyl-beta-cyclodextrin as a chiral selector, bonds the chiral selector on the inner wall of a capillary by a chemical bonding method, and realizes the separation of pantoprazole racemate by a capillary electrophoresis apparatus. The chiral capillary electrochromatographic column prepared by the invention has high separation efficiency, and the chiral selector is cheap and easy to obtain and has small dosage.

Drawings

FIG. 1 is an electrochromatography of separation of pantoprazole racemate according to the invention.

Detailed Description

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

FIG. 1 is a capillary electrochromatography for separating pantoprazole racemate, the separation and analysis time is within 17 minutes, and the separation degree is more than 7.

The first embodiment is as follows:

(1) intercept an uncoated Quartz chromatography column (inner diameter 75)

) The column length is 53 cm, and the effective length is 45 cm. Washing with pure methanol for 10 min; washing with distilled water for 30 min; washing with 1M NaOH for 1 h; washing with distilled water for 30 min; washing with absolute ethyl alcohol for 1 h; washing with 1% 3-aminopropyl-3-methoxysilane solution for 1 hr, sealing the column overnight, and oven drying in an oven at 100 deg.C for 24 hr.

The flushing flow rate is 800 mu L/h

(3) The electrochromatography separation conditions are as follows: the background electrolyte was 50 mM tris-H₃PO₄Solution, pH 5.0; the separation voltage is 15 kV; ultraviolet detection wavelength: 280 nm.

(4) Accurately weighing pantoprazole racemate 10 mg, placing the pantoprazole racemate in a 10 mL brown volumetric flask, dissolving the pantoprazole racemate in 10% NaOH solution, fixing the volume to a scale, and shaking up to obtain a test solution with the sample concentration of 1 mg/mL. Through a 0.45

Filtering with microporous membrane for use. The capillary column is washed by 0.1 mol/L NaOH, secondary distilled water and running buffer solution for 10min respectively, then baseline running is carried out, siphon sampling is adopted after the baseline is stable (about 2 min), the sampling height is 10 cm, the sampling time is 10 s, the anode sampling and the cathode detection are carried out, and an electrophoresis chart is recorded. The separation electrochromatography of pantoprazole racemate is shown in the attached figure.

Claims

1. An open-tube capillary electrochromatography method for separating pantoprazole racemate is characterized in that 6-tosyl-beta-cyclodextrin is used as a chiral selector and is bonded to the inner wall of a capillary by a chemical bonding method to prepare a chiral capillary electrochromatography column; separating pantoprazole enantiomers by a capillary electrophoresis apparatus;

the specific separation steps are as follows:

a, a column manufacturing mode:

(1) intercepting an uncoated quartz chromatographic column with the inner diameter of 75m, wherein the length of the column is 53 cm, burning the column at the position of 8 cm, wiping the column by using acetone until the column is completely transparent, and taking the column as a detection window, wherein the effective length of the column is 45 cm; washing with pure methanol for 10 min; washing with distilled water for 30 min; washing with 1M NaOH for 1 h; washing with distilled water for 30 min; washing with absolute ethyl alcohol for 1 h; washing with 1% 3-aminopropyl-3-methoxysilane solution for 1 hr, sealing the column overnight, and oven drying in an oven at 100 deg.C for 24 hr;

(2) washing the dried capillary tube with 1% 6-p-toluenesulfonyl-beta-CD-acetonitrile solution for 3 h, and standing for 1 h; then washing with 50% anhydrous ethanol for 10min, and then washing with 100% anhydrous ethanol for 30 min;

the flushing flow rate is 800 mu L \ h;

b. the electrochromatography separation conditions are as follows:

the background electrolyte was 50 mM tris-H₃PO₄Solution, pH 5.0; the separation voltage is 15 kV; ultraviolet detection wavelength: 280 nm; washing with 0.1M NaOH, redistilled water and running buffer solution for 10min before each analysis; washing the sample injection room with running buffer solution for 5 min, and then carrying out next sample injection;

the open-tube capillary electrochromatography separation condition is as follows: the buffer salt is 50 mmol/Ltris-H₃PO₄pH 5.0; the separation voltage is 15 kV; the ultraviolet detection wavelength is 280 nm.