Method for detecting content of isomers in Cbz-Glu

Abstract

The invention discloses a method for detecting isomer content in Cbz-Glu, belongs to the field of chemical analysis, and particularly relates to high performance liquid chromatography conditions as follows: a chromatographic column is CHIRALCEL OX-3, the detection wavelength is 210 nm, the column temperature is 35 DEG C, a mobile phase is a mixed solution of n-hexane absolute ethyl alcohol, the flow velocity of the mobile phase is 0.6 ml / min, and the elution time is 40 min. According to the method, the CbzL-Glu and the isomer Cbz-D-Glu of the CbzL-Glu can be effectively separated, so that thecontent of the isomer Cbz-D-Glu is detected in time.

Description

technical field

[0001] The invention belongs to the field of chemical analysis, in particular to a method for detecting the content of isomers in Cbz-Glu. Background technique

[0002] There are two isomers of Cbz-Glu, Cbz-L-Glu and Cbz-D-Glu, wherein Cbz-L-Glu is the target product, and Cbz-D-Glu is an impurity that is isomers of each other. During the production process, the content of Cbz-D-Glu needs to be strictly controlled, but there is no method for detecting the content of isomer Cbz-D-Glu impurities in Cbz-Glu in the prior art. Contents of the invention

[0003] In order to solve the above problems, the present invention provides a method for detecting the content of isomers in Cbz-Glu. The method of the invention can effectively separate the magazine Cbz-D-Glu, thereby measuring its content.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] A method for detecting the content of isomers in Cb...

Examples

Embodiment 1

[0020] (1) Set the HPLC conditions

[0021] The chromatographic column is CHIRALCEL OX-3, the specification of the chromatographic column is 250×4.6mm, 3μm, the detection wavelength is 210nm, the column temperature is 35°C, the mobile phase is a mixture of n-hexane-absolute ethanol, and the flow rate of the mobile phase is 0.6ml / min, the elution time was 40min, the injection volume was 10μL, the volume ratio of n-hexane to absolute ethanol in the mobile phase was 85:15, and 0.1% TFA was added to the mobile phase.

[0022] (2) Take 5mg of the test sample, weigh it accurately, put it in a 10mL volumetric flask, dissolve it with a diluent and dilute to the mark, and shake it well; the diluent is a mixture of n-hexane and absolute ethanol, and n-hexane The volume ratio with absolute ethanol is 80:20;

[0023] (3) Enter the obtained sample dilution into a high-performance liquid chromatography system for detection, and record the peak area of ​​Cbz-L-Glu and the peak area of ​​Cb...

Embodiment 2

[0027] (1) Set the HPLC conditions:

[0028] The chromatographic column is CHIRALCEL OX-3, the specification of the chromatographic column is 250×4.6mm, 3μm, the detection wavelength is 205nm, the column temperature is 35°C, the mobile phase is a mixture of n-hexane-absolute ethanol, and the flow rate of the mobile phase is 0.6ml / min, the elution time was 30min, the injection volume was 10μL, the volume ratio of n-hexane to absolute ethanol in the mobile phase was 85:15, and 0.1% TFA was added to the mobile phase.

[0029] (2) Take 5mg of the test sample, weigh it accurately, put it in a 10mL volumetric flask, dissolve it with a diluent and dilute to the mark, and shake it well; the diluent is a mixture of n-hexane and absolute ethanol, and n-hexane The volume ratio with absolute ethanol is 80:20;

[0030] (3) Enter the obtained sample dilution into a high-performance liquid chromatography system for detection, and record the peak area of ​​Cbz-L-Glu and the peak area of ​​C...

Embodiment 3

[0034] (1) Set the HPLC conditions:

[0035] The chromatographic column is CHIRALCEL OX-3, the specification of the chromatographic column is 250×4.6mm, 3μm, the detection wavelength is 220nm, the column temperature is 35°C, the mobile phase is a mixture of n-hexane-absolute ethanol, and the flow rate of the mobile phase is 0.6ml / min, the elution time was 35min, the injection volume was 10μL, the volume ratio of n-hexane to absolute ethanol in the mobile phase was 85:15, and 0.1% TFA was added to the mobile phase.

[0036] (2) Take 5mg of the test sample, weigh it accurately, put it in a 10mL volumetric flask, dissolve it with a diluent and dilute to the mark, and shake it well; the diluent is a mixture of n-hexane and absolute ethanol, and n-hexane The volume ratio with absolute ethanol is 80:20;

[0037] (3) Enter the obtained sample dilution into a high-performance liquid chromatography system for detection, and record the peak area of ​​Cbz-L-Glu and the peak area of ​​C...

technical field

[0001] The invention belongs to the field of chemical analysis, in particular to a method for detecting the content of isomers in Cbz-Glu. Background technique

[0002] There are two isomers of Cbz-Glu, Cbz-L-Glu and Cbz-D-Glu, wherein Cbz-L-Glu is the target product, and Cbz-D-Glu is an impurity that is isomers of each other. During the production process, the content of Cbz-D-Glu needs to be strictly controlled, but there is no method for detecting the content of isomer Cbz-D-Glu impurities in Cbz-Glu in the prior art. Contents of the invention

[0003] In order to solve the above problems, the present invention provides a method for detecting the content of isomers in Cbz-Glu. The method of the invention can effectively separate the magazine Cbz-D-Glu, thereby measuring its content.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] A method for detecting the content of isomers in Cb...

Examples

Embodiment 1

[0020] (1) Set the HPLC conditions

[0021] The chromatographic column is CHIRALCEL OX-3, the specification of the chromatographic column is 250×4.6mm, 3μm, the detection wavelength is 210nm, the column temperature is 35°C, the mobile phase is a mixture of n-hexane-absolute ethanol, and the flow rate of the mobile phase is 0.6ml / min, the elution time was 40min, the injection volume was 10μL, the volume ratio of n-hexane to absolute ethanol in the mobile phase was 85:15, and 0.1% TFA was added to the mobile phase.

[0022] (2) Take 5mg of the test sample, weigh it accurately, put it in a 10mL volumetric flask, dissolve it with a diluent and dilute to the mark, and shake it well; the diluent is a mixture of n-hexane and absolute ethanol, and n-hexane The volume ratio with absolute ethanol is 80:20;

[0023] (3) Enter the obtained sample dilution into a high-performance liquid chromatography system for detection, and record the peak area of ​​Cbz-L-Glu and the peak area of ​​Cb...

Embodiment 2

[0027] (1) Set the HPLC conditions:

[0028] The chromatographic column is CHIRALCEL OX-3, the specification of the chromatographic column is 250×4.6mm, 3μm, the detection wavelength is 205nm, the column temperature is 35°C, the mobile phase is a mixture of n-hexane-absolute ethanol, and the flow rate of the mobile phase is 0.6ml / min, the elution time was 30min, the injection volume was 10μL, the volume ratio of n-hexane to absolute ethanol in the mobile phase was 85:15, and 0.1% TFA was added to the mobile phase.

[0029] (2) Take 5mg of the test sample, weigh it accurately, put it in a 10mL volumetric flask, dissolve it with a diluent and dilute to the mark, and shake it well; the diluent is a mixture of n-hexane and absolute ethanol, and n-hexane The volume ratio with absolute ethanol is 80:20;

[0030] (3) Enter the obtained sample dilution into a high-performance liquid chromatography system for detection, and record the peak area of ​​Cbz-L-Glu and the peak area of ​​C...

Embodiment 3

[0034] (1) Set the HPLC conditions:

[0035] The chromatographic column is CHIRALCEL OX-3, the specification of the chromatographic column is 250×4.6mm, 3μm, the detection wavelength is 220nm, the column temperature is 35°C, the mobile phase is a mixture of n-hexane-absolute ethanol, and the flow rate of the mobile phase is 0.6ml / min, the elution time was 35min, the injection volume was 10μL, the volume ratio of n-hexane to absolute ethanol in the mobile phase was 85:15, and 0.1% TFA was added to the mobile phase.

[0036] (2) Take 5mg of the test sample, weigh it accurately, put it in a 10mL volumetric flask, dissolve it with a diluent and dilute to the mark, and shake it well; the diluent is a mixture of n-hexane and absolute ethanol, and n-hexane The volume ratio with absolute ethanol is 80:20;

[0037] (3) Enter the obtained sample dilution into a high-performance liquid chromatography system for detection, and record the peak area of ​​Cbz-L-Glu and the peak area of ​​C...

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