Method for qualitatively and quantitatively analyzing chiral substance based on circular dichroism spectrum technology

A spectroscopic technique and a 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 difficult popularization and utilization of detection methods, easy contamination of original samples, cumbersome operation, etc., and achieve pollution-free recovery of samples , simple calculation and easy operation

Active Publication Date: 2021-03-02
NINGBO UNIV
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  • Description
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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

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  • Method for qualitatively and quantitatively analyzing chiral substance based on circular dichroism spectrum technology
  • Method for qualitatively and quantitatively analyzing chiral substance based on circular dichroism spectrum technology
  • Method for qualitatively and quantitatively analyzing chiral substance based on circular dichroism spectrum technology

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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 qualitatively and quantitatively analyzing a chiral substance based on a circular dichroism spectrum technology. The method which comprises the following steps: 1), quickly establishing a first standard curve by using the circular dichroism spectrum technology to obtain a concentration difference value of a levorotatory compound and a dextrorotatory compound inthe chiral substance; 2), establishing a second standard curve by utilizing a high performance liquid chromatography technology to obtain the concentration sum value of the levorotatory compound andthe dextrorotatory compound in the chiral substance; and 3), finally, obtaining the concentrations of the levorotatory compound and the dextrorotatory compound in the chiral substance through simple calculation, and carrying out qualitative and quantitative analysis. The method has the advantages that the concentration of the enantiomer in the chiral substance can be quickly determined without adding a chiral reagent, in-situ qualitative and quantitative analysis and pollution-free sample recovery are realized, the operation is simple and convenient, the cost is low, the calculation is simple,and particularly, the chiral purity of each batch of crude drugs can be quickly analyzed in the field of chiral crude drug detection; and suitable popularization and utilization are realized.

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 Applications(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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