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Nucleoside and base group type substance separation method and application thereof

A separation method and base technology, which is applied in the field of separation of nucleosides and bases, can solve problems such as poor peak shape, low sensitivity, and harsh chromatographic conditions, and achieve good peak shape, simple operation, and high sensitivity.

Pending Publication Date: 2018-05-15
SPH NO 1 BIOCHEM & PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is that the existing methods for separating and purifying nucleosides and bases are complicated, complex samples are difficult to achieve effective separation, chromatographic conditions are harsh, long-term use will cause large damage to the chromatographic column, poor peak shape, and low sensitivity , poor repeatability and other problems, thus providing a separation method and application of nucleoside and base substances

Method used

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  • Nucleoside and base group type substance separation method and application thereof
  • Nucleoside and base group type substance separation method and application thereof
  • Nucleoside and base group type substance separation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1. Preparation of test samples

[0044] Take an appropriate amount of each reference substance of adenosine, cytidine, guanosine, adenine, hypoxanthine, and guanine, weigh them accurately, add water to make a stock solution with a concentration of 1 mg / ml, and accurately draw an appropriate amount of each stock solution of the reference substance in In the same measuring bottle, add water to dilute to the scale to obtain a mixed solution with a concentration of 1 μg / ml, and dilute it with acetonitrile to obtain a mixed reference solution of nucleosides and bases.

[0045] 2. Chromatographic conditions and methods

[0046] Liquid chromatograph: Thermo-Fisher UltiMate 3000series high performance liquid chromatograph

[0047] Chromatographic column: ACQUITYXBridge Amide (100mm×2.1mm, 3.5μm)

[0048] Mobile phase: Acetonitrile (containing 0.1% formic acid) and water (containing 5 mM ammonium formate, 5 mM ammonium acetate, 0.1% formic acid) were used as mobile phases A an...

Embodiment 2

[0062] 1. Preparation of test samples

[0063] Take Gualoupi Injection (produced by Shanghai Shangyao No.1 Biochemical Pharmaceutical Co., Ltd.) 1ml, add water to dilute to 50ml, mix well, dilute twice with acetonitrile to get ready.

[0064] 2. Chromatographic conditions and methods

[0065] With embodiment 1.

[0066] 3. Test results

[0067] The retention time of adenosine is 2.975min, the retention time of cytidine is 6.154min, the retention time of guanosine is 7.045min, the retention time of adenine is 3.512min, the retention time of hypoxanthine is 4.631min, the retention time of guanine The time is 5.413min. The resolution is greater than 1.5, and the peak shape is good without tailing or protruding.

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Abstract

The invention discloses a nucleoside and base group type substance separation method and application thereof. A high performance liquid chromatography is used for separating nucleoside and base grouptype substances; a chromatographic column in the high performance liquid chromatography is an amino chromatographic column of a triple-bond bonded acylamino bonded phase; mobile phases in the high performance liquid chromatography are a saline water containing solution and a polar organic solvent; the nucleoside and base group type substances are adenosine, cytidine, guanosine, adenine, hypoxanthine and guanine. The separation method has the advantages that the operation is simple; six kinds of nucleoside and base group type substances can be completely separated; the peak shape is good; the interference by other organic impurities is avoided; the sensitivity is high; simplicity and high speed are realized.

Description

technical field [0001] The invention relates to a method for separating nucleosides and base substances and its application. Background technique [0002] In the past 20 years, with the advancement of biotechnology, nucleic acid series products, including nucleotides, nucleosides and their derivatives, have been widely used. Among them, nucleoside compounds have shown irreplaceable and unique effects in antiviral and antitumor aspects. Most of the anti-AIDS drugs approved by the US FDA are nucleoside derivatives. Nucleoside drugs are expected to become a new generation of important drugs after sulfonamides and antibiotics. [0003] For a class of new drugs, the research and monitoring of their metabolites is particularly important, especially the metabolites of nucleoside drugs are endogenous substances. Adenine, guanine, cytosine, and uracil are the end products of ribonucleic acid (RNA) metabolism. Adenine, guanine, cytosine, and thymine are the final products of deoxy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/72B01D15/08
CPCB01D15/08G01N30/02G01N30/72
Inventor 季申胡青黄臻辉孙健冯睿于泓毛丹朱玲玲
Owner SPH NO 1 BIOCHEM & PHARMA CO LTD
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