Method for enhancing stability of amino column

An amino column and stability technology, applied in the field of analytical chemistry, can solve the problems of high baseline noise, poor separation selectivity, time-consuming, etc., and achieve the effects of good reproducibility, enhanced use stability, and development promotion.

Pending Publication Date: 2021-02-02
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One is to improve the filler preparation process, such as TSKgel NH from TOSOH 2 -100 and Welch's Ultimate XB-NH 2 , but these two amino columns still have high baseline noise and poor separation selectivity in the ELSD analysis of sorbitol and mannitol, and the poor separation and analysis results are derived from the instability of the amino column sex
The second is to use a suitable mobile phase chromatographic column to flush, but the method of isopropanol flushing in the European Pharmacopoeia is not only time-consuming (continuous flushing for 10 hours at a low flow rate), but also cannot be used in the ELSD analysis and detection of sorbitol and mannitol. get satisfactory results

Method used

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  • Method for enhancing stability of amino column
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  • Method for enhancing stability of amino column

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] First, prepare 20mM phosphate solution (including sodium dihydrogen phosphate 10mM, disodium hydrogen phosphate 10mM); then, mix the phosphate solution with methanol in equal volume as the mobile phase to wash R5-NH 2 Column, wash 18 column volumes at a flow rate of 0.5mL / min; finally, wash R5-NH with water / methanol solution with a volume ratio of water to methanol of 1 / 1 as the mobile phase 2 The column was flushed for 6 column volumes at a flow rate of 0.5 mL / min.

[0021] Taking uracil and uridine as analysis object, adopt the method of the present invention to R5-NH 2 Column R5-NH before and after flushing 2 Column (4.6×150mm) analysis of uracil, uridine reproducibility comparison example figure 1 shown.

[0022] Depend on figure 1 As can be seen, the method of the present invention is not used to flush R5-NH 2 Before column, R5-NH 2 The reproducibility of the continuous injection of the column for 5 times is very poor; however, after washing by the present in...

Embodiment 2

[0024] The difference from Example 1 is that 48 column volumes are washed with phosphate / methanol solution; finally, 12 column volumes are washed with water / methanol solution.

[0025] Take sorbitol, mannitol as analysis object, adopt the inventive method to R5-NH 2 Column R5-NH before and after flushing 2 Comparison of column (4.6×150mm) separation and analysis effects on sorbitol and mannitol figure 2 shown.

[0026] Depend on figure 2 It can be seen that R5-NH 2 The baseline noise is strong and the separation selectivity is poor before column flushing; but after flushing by the method of the invention, the baseline noise is obviously weakened and the separation selectivity is obviously improved. The above results show that the present invention can reduce the baseline noise when using an evaporative light scattering detector and provide better separation selectivity while rapidly and effectively enhancing the stability of the amino column.

Embodiment 3

[0028] The difference from Example 2 is that other different brands of amino columns (4.6 × 150mm) are used to compare the separation and analysis effects of other different brands of amino columns (4.6 × 150mm) to sorbitol and mannitol before and after washing with the method of the present invention. Comparison picture as image 3 shown.

[0029] Depend on image 3 It can be seen that the method for enhancing the stability of the amino column of the present invention is universal, that is, the present invention is not only applicable to a certain type of amino column, but also applicable to other brands of amino columns.

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Abstract

The invention provides a method for enhancing the stability of an amino column. The method can obviously enhance the use stability and repeatability of the amino column. According to the method, the phosphate/methanol solution is used as a mobile phase to wash the amino column, and then the water/methanol solution is used as a mobile phase to wash the amino column so that the surface of the columnis stabilized, and good use stability is obtained. According to the method for enhancing the stability of the amino column, good reproducibility of separation analysis based on the amino column can be obtained, baseline noise generated when an evaporative light scattering detector is used is reduced, better separation selectivity is provided, and the method has very important value on applicationof the amino column and is also an important direction of chromatographic research.

Description

technical field [0001] The invention belongs to the field of analytical chemistry and relates to a method for enhancing the stability of an amino column. [0002] technical background [0003] Amino columns are widely used in chromatographic separation and analysis, such as sugars and certain polar drugs, pharmaceutical excipients, etc. However, amino columns have problems such as poor stability and reproducibility, and short lifetime. Although the amino column has the above disadvantages, and new HILIC stationary phases are constantly being introduced, the amino column cannot be completely replaced due to its unique selectivity. Therefore, enhancing the stability of the amino column is of great value to its application, and it is also an important direction of chromatography research. [0004] At present, the methods to enhance the stability of amino columns mainly start from two aspects. One is to improve the filler preparation process, such as TSKgel NH from TOSOH 2 -1...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/60G01N30/74G01N30/88G01N30/32G01N30/34
CPCG01N30/32G01N30/34G01N30/60G01N30/74G01N30/88G01N2030/324G01N2030/889
Inventor 梁鑫淼夏东海郭志谋金高娃闫竞宇
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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