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A Qualitative and Quantitative Analysis Method of Chiral Substances Based on Circular Dichroism Spectroscopy

A spectroscopic technique and quantitative analysis technique, applied in the field of qualitative and quantitative analysis of chiral substances based on circular dichroism spectroscopy, can solve the problems of easy contamination of original samples, cumbersome operation, complex calculation, etc., and the method is fast, simple and universal The effect of high reliability and low instrument cost

Active Publication Date: 2022-03-29
NINGBO UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The HPLC-CD method is to combine circular dichroism spectrometer and high-performance liquid chromatography to measure circular dichroism spectrum signal and chromatographic peak area respectively, and obtain the purity of enantiomers in the chiral substance through calculation; however prior art The detection method still has the problems of requiring additives as internal standards, easily contaminating the original sample, cumbersome operation, and complicated calculations, especially in the detection field of chiral APIs, which often need to quickly analyze the chiral purity of each batch of APIs. Existing detection methods are difficult to popularize and utilize

Method used

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  • A Qualitative and Quantitative Analysis Method of Chiral Substances Based on Circular Dichroism Spectroscopy
  • A Qualitative and Quantitative Analysis Method of Chiral Substances Based on Circular Dichroism Spectroscopy
  • A Qualitative and Quantitative Analysis Method of Chiral Substances Based on Circular Dichroism Spectroscopy

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Embodiment 1

[0044] Determination of L-leucine (L-Leu) Concentration by Circular Dichroism Spectroscopy

[0045] Accurately weigh 26.23mg of L-leucine (L-Leu), add 2mL of distilled water or deionized water as a stock solution; then dilute to concentrations of 0.005M, 0.01M, 0.015M, 0.025M standard solution, the circular dichroism chromatograms of each L-Leu standard solution are obtained by circular dichroism detection, such as figure 1 As shown, it is found that L-Leu has a circular dichroism peak with a positive signal at a wavelength of 200 nm.

[0046] Take the ellipticity θ of the circular dichroism peak at the wavelength of 200nm respectively, and draw a linear standard relationship curve with the corresponding concentration c, as figure 2 As shown, the fitting equation y=0.0002x-0.0005 of the ellipticity θ of the circular dichroism peak at the wavelength of 200nm and the concentration c is obtained, wherein, R 2 =0.9993, indicating that the degree of linear correlation between θ ...

Embodiment 2

[0053] Concentration Determination of Components of Leu's Enantiomer Mixture

[0054] There are two kinds of enantiomers of Leu, i.e. the left-handed compound L-Leu and the right-handed compound D-Leu; the absorption peaks of L-Leu and D-Leu on the achiral chromatographic column are overlapped, using the examples The L-Leu standard solution of five kinds of concentration in one carries out the determination of achiral HPLC, wherein the achiral HPLC technology is the mobile phase that does not use chiral chromatographic column and does not contain chiral additive, draws the wavelength at 210nm place The linear relationship curve graph of signal peak area integral value and concentration, such as image 3 As shown, the fitting equation y=4E-5x-0.001 is obtained, where R 2 =0.9991, indicating that the linear correlation between the integral value of the signal peak area and the concentration is good.

[0055] Concentration determination of mixture A: Take L-Leu and D-Leu with a...

Embodiment 3

[0065] The high-performance liquid chromatograph of embodiment two is also connected with an ultraviolet detector, obtains the absorbance of the L-Leu standard solution of five kinds of concentrations in embodiment one at a wavelength of 210nm, draws the absorbance of each L-Leu standard solution and the corresponding Concentration linear relationship graph, obtain the fitting equation of absorbance and concentration, all the other are identical with embodiment two.

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Abstract

The invention discloses a method for qualitative and quantitative analysis of a chiral substance based on a circular dichroism spectroscopy technique, comprising the following steps: 1) using the circular dichroism spectroscopy technique to rapidly establish a first standard curve to obtain the levorotatory compound and the chiral substance in the chiral substance. 2) Use high performance liquid chromatography to establish a second standard curve to obtain the concentration and value of the levorotatory and dextrorotatory compounds in the chiral substance; 3) Finally, obtain the concentration of the chiral substance by simple calculation The concentration of levorotatory compounds and dextrorotatory compounds is used for qualitative and quantitative analysis; the advantage of the present invention is that the concentration of enantiomers in chiral substances can be quickly determined without external chiral reagents, and in-situ qualitative and quantitative analysis is realized. The sample is recovered without pollution, and the operation is simple, the cost is low, and the calculation is simple. Especially in the field of detection of chiral APIs, the chiral purity of each batch of APIs can be quickly analyzed, which is suitable for popularization and utilization.

Description

technical field [0001] The invention belongs to the technical field of chemical detection, in particular to a qualitative and quantitative analysis method for chiral substances based on circular dichroism spectroscopy. Background technique [0002] There are a large number of chiral substances in nature, including many biomolecules and drugs in living organisms. The raw materials used to synthesize proteins in organisms are all L-type amino acids, and the nucleosides used to synthesize nucleic acids are all D-type, while the corresponding raw materials with opposite chirality cannot be used to synthesize proteins or nucleic acids; in the field of medicine, as people The research on chiral drugs continues to deepen, and it is gradually realized that different isomers of chiral drugs, especially enantiomers, have different or even opposite pharmacological and toxicological effects. Therefore, the development of qualitative and quantitative analysis methods for chiral substanc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/19G01N30/02G01N30/74G01N30/86
CPCG01N21/19G01N30/02G01N30/74G01N30/8675
Inventor 应见喜赵玉芬丁瑞文
Owner NINGBO UNIV
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