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Mobile phase for separating phospholipids in high efficiency liquid chromatography

A high-performance liquid chromatography and mobile phase technology, applied in the field of organic chemical analysis, can solve problems such as system leakage, achieve the effects of good use, increase pH, and reduce damage

Active Publication Date: 2008-04-09
西安血氧生物技术有限公司
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Problems solved by technology

Among them, the phosphoric acid / sulfuric acid in the mobile phase composed of acetonitrile-methanol-phosphoric acid / sulfuric acid will produce precipitation, which will cause scratches on the plunger and gasket of the high-pressure pump in the high-pressure liquid chromatography, resulting in system leakage

Method used

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  • Mobile phase for separating phospholipids in high efficiency liquid chromatography
  • Mobile phase for separating phospholipids in high efficiency liquid chromatography
  • Mobile phase for separating phospholipids in high efficiency liquid chromatography

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Embodiment Construction

[0028] The specific implementation and effect of the distribution ratio of the components of the mobile phase of the present invention will be described below in conjunction with the accompanying drawings, and the specific chromatographic conditions for the separation and detection of the mobile phase of the present invention will be described.

[0029] High performance liquid chromatography includes system controller, ultraviolet detector, high performance liquid phase pump system, degassing system and column thermostat. The chromatographic column is a reversed-phase analytical column C18 (250mm×4.6mm I.D.), the drift tube temperature of the evaporative light scattering detector is 80°C, the nitrogen flow rate is 2L / min; the mobile phase flow rate is 1.0ml / min.

[0030] Separation and detection experiment initially uses HPLC-UV to separate and detect phospholipids. The sample is the phospholipids extracted from porcine erythrocyte membranes. The volume ratio of the mobile phas...

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Abstract

The invention relates to a mobile phase for separating phosphatide by a highly effective liquid phase chromatography. The components of the mobile phase comprise 10 percent to 50 percent of methyl alcohol, 50 percent to 85 percent of acetonitrile, 0.5 percent to 3 percent of fluorinated organic acid, and 0.5 percent to 3 percent of water. Through the long-term operation, the mobile phase of the invention cannot result in any deposit, cannot damage a pump system, and has no damage to the pump system of a liquid phase chromatographic instrument; the mobile phase can be used to detect the separated phosphatide in an evaporative light-scattering detector, compared with an ultraviolet detector, the base line detected by the evaporative light-scattering detector is stable, the quantitative analysis is easy, the sensitivity is high, and the application of the mobile phase has no limit; compared with the gradient method, the method of the invention uses a single pump to realize the separating detection; when the mobile phase adopts the appropriate compounding ratio, besides the phosphatide can be separated, the unknown materials that cannot be separated by the other mobile phases can also be separated. When the mobile phase of the invention applies to the isocratic eluting, the operation is simple and time-saving, the reproducibility is good, and the duplicity is easy.

Description

technical field [0001] The invention relates to a mobile phase used in the separation of phospholipids by high performance liquid chromatography, and belongs to the technical field of organic chemical analysis. Background technique [0002] Methods for separating phospholipids by high performance liquid chromatography include isocratic elution and gradient elution. Generally use high-performance liquid chromatography and ultraviolet detector combination (HPLC-UV) to detect phospholipids, high-performance liquid chromatography provides the mobile phase with stable flow rate and higher pressure to the chromatographic column, so that the sample is separated at the chromatographic column, and then The separated phospholipids were detected with a UV detector. If the proportion of each solvent in the mobile phase is constant during detection, it is an isocratic elution method, and if the instrument automatically mixes different solvents according to requirements, it is a gradient...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/08
Inventor 严坤平但宁陈超
Owner 西安血氧生物技术有限公司
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