High performance liquid chromatography separating column suitable for amino acid chiral resolution

A high-performance liquid chromatography and chiral separation technology, which is applied in the field of high-performance liquid chromatography separation columns for chiral separation of amino acids, can solve problems such as harsh temperature conditions, no separation effect, and no solution, and achieve separation Fast speed, low cost of column making, and high reproducibility

Inactive Publication Date: 2015-04-01
YUNNAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of chiral HPLC columns to separate chiral drugs is one of the most effective methods, but the current HPLC separation columns specially used for chiral resolution of amino acids still have the following problems, such as: the chirality of mixed amino acids The resolution effect is not good, the temperature conditi

Method used

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  • High performance liquid chromatography separating column suitable for amino acid chiral resolution
  • High performance liquid chromatography separating column suitable for amino acid chiral resolution
  • High performance liquid chromatography separating column suitable for amino acid chiral resolution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The method for synthesizing R-(3,3'-dibromo-1,1'-dinaphthyl)-20-crown-6 (such as figure 1 Shown): R-binaphthol 10g is added in 180mL acetonitrile, adds 20g K 2 CO 3 and 0.5g CsCO 3 As a catalyst, stir under magnetic reflux at 70°C, slowly add 6.52 mL of methyl iodide dropwise during the reflux process, and react for 4 hours. The mixture was extracted with dichloromethane, the organic layer was dried and purified by silica gel column [eluent V (ethyl acetate): V (petroleum ether) = 1:5], and the eluent was removed under reduced pressure to obtain 10.54 g of white crystal B; Under the protection of nitrogen, 37.6mL of normal C 4 h 9 Li (1.6mol / L) was added to a solution of tetramethyldiethylamine (6.1mL) in 200mL of ether, stirred at 25°C for 15min, and 8.0g of product B was added and stirred for 3h. Then the reactant was cooled to -75°C, and Br was added within 10 min. 2 (3.9mL) in pentane (20mL) solution, then the reaction was raised to room temperature and stirre...

Embodiment 2

[0023] With the chromatographic column obtained in Example 1, the detection wavelength of the ultraviolet detector is 210nm, the mobile phase is pH=2, and the perchloric acid solution that is ultrasonically degassed is filtered through a 0.45 μm filter membrane, and the flow rate is 0.5mL min -1 , under the condition that the column temperature of the liquid phase column is 25°C, test the liquid chromatography chiral resolution effect on DL-glutamine and DL-asparagine, see image 3 .

[0024] Depend on image 3 It can be seen that the chromatographic column obtained in Example 1 can achieve baseline separation for DL-glutamine and partial separation for DL-asparagine, indicating that it also has a certain separation effect for non-protein α-chiral primary amines.

Embodiment 3

[0026] With the chromatographic column of the present invention a (Embodiment 1 gained) and commodity CR (+) column b The chiral column detects 21 kinds of α-amino acid enantiomers (the first 19 are amino acids that make up proteins, and the latter two are other chiral primary amines). The detection wavelength of the ultraviolet detector is 210nm, and the mobile phase is pH=1. Filter the ultrasonically degassed perchloric acid solution with a μm filter membrane at a flow rate of 0.5mL min -1 , the chromatographic chiral resolution effect under the condition that the column temperature of the liquid phase column is 25°C. retention factor k 1 =(t 1 -t 0 ) / t 0 ,k 2 =(t 2 -t 0 ) / t 0 ;Separation factor α=k 2 / k 1 ; Resolution Rs=1.18(t 2 -t 1 ) / (W 1 / 2 (1)+W 1 / 2 (2)); where t 1 , t 2 Respectively, the retention time of the first elution peak and the second elution peak of the sample; W 1 / 2 (1), W 1 / 2 (2) are respectively the half-peak widths of two enantiomer chro...

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Abstract

The invention discloses a high performance liquid chromatography separating column suitable for amino acid chiral resolution. The separating column is prepared by the following steps: dissolving R(or S)-(3,3'-bromo-1,1'-dinaphthyl)-20-crown-6 in dichloromethane, uniformly dispersing the crown ether solution to C18 silica gel, carrying out rotary evaporation to remove the solvent to prepare a chiral stationary phase; and mixing the chiral stationary phase in a methanol water solution, stirring to obtain a homogenate solution, and filling the chromatographic column by a wet process by using the methanol water solution as a displacement fluid. The separating column can effectively separate all the 19 protein amino acids with chiral site at normal temperature, and can effectively separate chiral drugs with primary amine on the chiral site. The separating column has the obvious characteristics of high resolving power, high separating rate, high reproducibility, lower preparation cost and the like, and can be used repeatedly. The separating column has obviously better separating effect for amino acids than the like products at home and abroad.

Description

technical field [0001] The invention belongs to the technical field of high-performance liquid chromatography separation, and in particular relates to a high-performance liquid chromatography separation column especially suitable for chiral separation of amino acids. Background technique [0002] High performance liquid chromatography has the advantages of high pressure, high speed, high efficiency, high sensitivity, wide application range, and no damage to samples. It has been widely used in nucleic acids, peptides, lactones, fused ring aromatics, surfactants, antioxidants, and medical drugs. analysis of other substances. The use of chiral HPLC columns to separate chiral drugs is one of the most effective methods, but the current HPLC separation columns specially used for chiral resolution of amino acids still have the following problems, such as: the chirality of mixed amino acids The resolution effect is not good, the temperature conditions are relatively harsh in the ch...

Claims

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Application Information

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IPC IPC(8): B01J20/29B01J20/30B01D15/22C07B57/00C07C227/34C07C229/08C07C229/24C07C229/36C07C229/22C07C323/58C07C319/28C07C279/14C07C227/08C07C237/06C07C227/40C07D207/16C07D233/64
CPCB01D15/22B01J20/29B01J2220/52B01J2220/80C07B57/00C07C227/34C07C277/08C07C319/28C07D207/16C07D233/64C07C229/08C07C229/24C07C229/36C07C229/22C07C227/40C07C279/14C07C323/58
Inventor 袁黎明伍鹏路振宇
Owner YUNNAN NORMAL UNIV
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