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High performance liquid chromatography detection methods for glycyl-D-glutamine

A technology of glutamine and high performance liquid phase, applied in the field of high performance liquid chromatography detection, can solve the problems of long test time, long reaction time, difficult separation, etc., and achieves the improvement of test efficiency, wide application range and good economic value. Effect

Active Publication Date: 2019-12-20
SHANDONG WEIGAO PHARM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] When developing a drug with an asymmetric center, it is necessary to use a variety of chiral chromatographic columns and chiral mobile phases for testing, which takes a long time to test and the effect is not ideal
Glycylglutamine is the terminal absorption, and the ultraviolet absorption is not strong. The separation and determination of glycylglutamine isomers using chiral chromatographic columns is not only costly, but also difficult to separate.
In the prior art, other products with weaker ultraviolet absorption have been reported to be derivatized by derivatization reagents and then separated by ordinary chromatographic columns. However, the method requires a long reaction time and requires steps such as solvent drying, redissolution, and filtration. , the operation steps are cumbersome

Method used

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  • High performance liquid chromatography detection methods for glycyl-D-glutamine
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  • High performance liquid chromatography detection methods for glycyl-D-glutamine

Examples

Experimental program
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Effect test

Embodiment 1

[0050] Chromatographic conditions: covalently bonded on the surface of silica gel-quinine (8R, 9S)-(1R, 2R)-cyclohexylsulfamic acid derivatives as a chromatographic column, and the mobile phase is formic acid-diethylamine-methanol ( 1.9:2.6:995.5), the detection wavelength is 248nm, the flow rate is 0.2ml / min, the column temperature is 20°C, and the injection volume is 2μl.

[0051]Preparation of the test solution: Accurately weigh an appropriate amount of glycyl-L-glutamine, dilute it with water to a solution containing 3mg of glycyl-L-glutamine per 1ml of water, as the test solution; Take appropriate amount of glycyl-L-glutamine and glycyl-D-glutamine, dilute with water so that every 1ml of water contains 3mg of glycyl-L-glutamine, glycyl-D-glutamine 4.5 μg of the mixed solution, as a mixed reference solution. Precisely measure 10 μl of the test solution, put it in a dry derivation tube (diameter 6 mm, height 50 mm), add 30 μl of buffer solution (Buffer1), vortex mix, add 6...

Embodiment 2

[0054] Chromatographic conditions: covalently bonded on the surface of silica gel-quinine (8R, 9S)-(1R, 2R)-cyclohexylsulfamic acid derivatives as a chromatographic column, mobile phase is formic acid-diethylamine-methanol ( 1.9:2.6:995.5), the detection wavelength is 248nm, the flow rate is 0.2ml / min, the column temperature is 22°C, and the injection volume is 2μl.

[0055] Preparation of the test solution: Accurately weigh an appropriate amount of glycyl-L-glutamine, dilute it with water to a solution containing 3mg of glycyl-L-glutamine per 1ml of water, as the test solution; Take appropriate amount of glycyl-L-glutamine and glycyl-D-glutamine, dilute with water so that every 1ml of water contains 3mg of glycyl-L-glutamine, glycyl-D-glutamine 4.5 μg of the mixed solution, as a mixed reference solution. Precisely measure 10 μl of the test solution, put it in a dry derivation tube (diameter 6 mm, height 50 mm), add 30 μl of buffer solution (Buffer1), vortex mix, add 60 μl of...

Embodiment 3

[0058] Chromatographic conditions: covalently bonded on the surface of silica gel-quinine (8R, 9S)-(1R, 2R)-cyclohexylsulfamic acid derivatives as a chromatographic column, mobile phase is formic acid-diethylamine-methanol ( 1.9:2.6:995.5), the detection wavelength is 248nm, the flow rate is 0.22ml / min, the column temperature is 20°C, and the injection volume is 2μl.

[0059] Preparation of the test solution: Accurately weigh an appropriate amount of glycyl-L-glutamine, dilute it with water to a solution containing 3mg of glycyl-L-glutamine per 1ml of water, as the test solution; Take appropriate amount of glycyl-L-glutamine and glycyl-D-glutamine, dilute with water so that every 1ml of water contains 3mg of glycyl-L-glutamine, glycyl-D-glutamine 4.5 μg of the mixed solution, as a mixed reference solution. Precisely measure 10 μl of the test solution, put it in a dry derivation tube (diameter 6 mm, height 50 mm), add 30 μl of buffer solution (Buffer1), vortex mix, add 60 μl o...

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Abstract

The invention discloses high performance liquid chromatography detection methods for glycyl-D-glutamine. A method comprises the following steps of (1), preparing a test solution; (2), preparing a mixed reference solution; (3), deriving the test solution; (4), deriving the mixed reference solution; and (5), carrying out chromatography, specifically, precisely measuring 2mul of each sample derived from the (3) and the (4), after chiral chromatographic columns are balanced, injecting the samples into liquid chromatographs, collecting chiral separation chromatograms, recording the chromatograms for 60min, and checking separation effects. According to the method, the chiral separation problem of glycyl-L-glutamine and the glycyl-D-glutamine is realized; testing efficiency is improved; stabilityof the chromatographic columns is relatively high; after separation is carried out for many times, the good separation effect also can be obtained; an application range is wide; and the method has good economic value.

Description

technical field [0001] The invention belongs to the technical field of high performance liquid chromatography detection and relates to a high performance liquid phase detection method of glycyl-D-glutamine. Background technique [0002] The presence of amino acids in the human body not only provides important raw materials for protein synthesis, but also provides a material basis for promoting growth, normal metabolism, and maintaining life. Amino acid drugs have low side effects, high drug efficacy, strong pertinence, and will not accumulate in the body to cause poisoning. [0003] Glycyl-L-glutamine is often used to provide glutamine to promote protein synthesis. The amino acids that make up proteins are all L-type amino acids, and the drugs obtained through chemical synthesis are often enantiomers, not a single optical Isomers, in many cases, there are significant differences in the pharmacological activity, metabolic process, metabolic rate and toxicity of a pair of ena...

Claims

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Application Information

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IPC IPC(8): G01N30/88G01N30/06
CPCG01N30/06G01N30/88G01N2030/067G01N2030/884
Inventor 杨万运王智军冷嘉琦罗艳玲韩瑞花姜吉慧
Owner SHANDONG WEIGAO PHARM CO LTD
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