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A kind of high performance liquid chromatography detection method of acetyl tyrosine and its optical isomers

A high-performance liquid chromatography and optical isomer technology, which is applied in the field of high-performance liquid chromatography detection of acetyltyrosine and its optical isomers, can solve the complex resolution mechanism of chiral mobile phase additives and the operation of derivatization reactions Complicated, interfering chromatographic separation and detection problems, to achieve the effect of simple method, high sensitivity and good reproducibility

Active Publication Date: 2017-01-04
WUHAN GRAND HOYO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the derivatization reaction operation is complicated, and the derivatization reagent and reaction medium may interfere with chromatographic separation and detection; the separation mechanism of chiral mobile phase additives is more complicated, the establishment of the method is more difficult, and the system takes a long time to balance when changing the mobile phase

Method used

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  • A kind of high performance liquid chromatography detection method of acetyl tyrosine and its optical isomers
  • A kind of high performance liquid chromatography detection method of acetyl tyrosine and its optical isomers
  • A kind of high performance liquid chromatography detection method of acetyl tyrosine and its optical isomers

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The separation effect test of different chiral columns of embodiment 1

[0028] 1. Instruments and conditions:

[0029] High performance liquid chromatography: Hitachi

[0030] Chromatographic column: the surface of silica gel is coated with cellulose-tris(3,5-xylylamino formate) (OD-H, Daicel4.6mm*250mm, 5μm); the surface of silica gel is coated with cellulose-tris( 4-methylbenzoate) (OJ-H, Daicel 4.6mm*250mm, 5μm); the surface of the silica gel was coated with amylose-tris[(S)-α-phenylethylcarbamate] (AS -H, Daicel 4.6mm*250mm, 5μm); the surface of silica gel is coated with amylose-tris(3,5-xylylcarbamate) (AD-H, Daicel 4.6mm*250mm, 5μm); silica gel The surface was coated with amylose-tris(5-chloro-2-methylphenylcarbamate) (AY, Guangzhou Yanchuang, 4.6 mm*250 mm, 5 μm).

[0031] Mobile phase: n-hexane-ethanol (80:20)

[0032] Flow rate: 1ml / min

[0033] Column temperature: 40°C

[0034] Inlet volume: 20μl

[0035] Detection wavelength: 210nm

[0036] 2. Experi...

Embodiment 2

[0042] The separation effect test of embodiment 2 different mobile phases

[0043] 1. Instruments and conditions:

[0044] High performance liquid chromatography: Hitachi

[0045] Chromatographic column: the surface of silica gel is coated with amylose-tris(5-chloro-2-methylphenylcarbamate) (AY, Guangzhou Yanchuang, 4.6mm*250mm, 5μm).

[0046] Mobile phase: n-hexane-ethanol (75:25; 80:20; 85:15)

[0047] Flow rate: 1ml / min

[0048] Column temperature: 40°C

[0049] Inlet volume: 20μl

[0050] Detection wavelength: 210nm

[0051] 2. Experimental methods and results:

[0052] Take each qualitative control solution and mixed standard solution, use n-hexane:ethanol=75:25, n-hexane:ethanol=80:20, n-hexane:ethanol=85:15 as mobile phase respectively, use AY chiral chromatographic column, press Above-mentioned chromatographic condition carries out HPLC analysis, record chromatogram, the result sees attached Figure 1-3 , the No. 1 peak in the figure is N-acetyl-D-tyrosine, and...

Embodiment 3

[0055] Determination of content of acetyltyrosine and its optical isomers in the sample of embodiment 3

[0056] 1. Instruments and conditions:

[0057] High performance liquid chromatography: Hitachi

[0058] Chromatographic column: the surface of silica gel is coated with amylose-tris(5-chloro-2-methylphenylcarbamate) (AY, Guangzhou Yanchuang, 4.6mm*250mm, 5μm)

[0059] Mobile phase: n-hexane-ethanol (85:15)

[0060] Flow rate: 1ml / min

[0061] Column temperature: 40°C

[0062] Inlet volume: 20μl

[0063] Detection wavelength: 210nm

[0064] 2. Detection methods and results:

[0065] 1) Preparation of the test solution: take 5 mg of the sample to be tested, dissolve it with 10 ml of absolute ethanol, and use it as the test solution;

[0066] 2) Preparation of the reference solution: take 2.5 mg of each of N-acetyl-L-tyrosine and its enantiomer N-acetyl-D-tyrosine, put it in a 10 mL volumetric flask, add absolute ethanol to dissolve And diluted to the mark, as the ref...

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Abstract

The invention discloses a high performance liquid chromatography detection method of acetyl tyrosine and an optical isomer thereof, and belongs to the technical field of pharmaceutical analysis and detection. The method comprises the step that acetyl tyrosine and the optical isomer thereof are separated and detected by using an amylase coated chiral chromatographic column and using a positive phase mixed solvent as a mobile phase. N- acetyl-L-tyrosine and N-acetyl-D-tyrosine can be effectively separated so that the content of the N- acetyl-L-tyrosine and the isomer thereof in a sample are effectively separated. The method disclosed by the invention is simple, convenient, reliable and fast, high in sensitivity, good in separation degree and reproducibility and accurate and reliable in result, and thus the method can be applied to quality control of the N- acetyl-L-tyrosine.

Description

technical field [0001] The invention belongs to the technical field of drug analysis and detection, and in particular relates to a high performance liquid chromatography detection method for acetyltyrosine and its optical isomers. Background technique [0002] N-acetyl-L-tyrosine is an important fine organic chemical intermediate, its chemical structure is: [0003] [0004] N-acetyl-L-tyrosine is widely used in medicine, pesticide, chemical industry and other fields. At present, only the routine physical and chemical indicators of N-acetyl-L-tyrosine are usually tested, but by-products will be produced in the production process. ——The optical isomer of N-acetyl-D-tyrosine, which affects product quality. Establishing a method that can accurately determine the content of acetyltyrosine and its optical isomers will have important guiding significance for its production. [0005] There are usually three methods for HPLC to resolve amino acid racemates: one is to use chiral...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/60
Inventor 黄蕾李建华
Owner WUHAN GRAND HOYO
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