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Mixed fatty acid high performance liquid phase chromatography method and application

A high-performance liquid chromatography and mixed fatty acid technology, which is applied in the direction of material separation, analysis materials, instruments, etc., can solve the problems of high absorbance, low reproducibility, complicated operation, etc., and achieve accurate and reliable results, simple and easy operation, and detection The effect of high sensitivity

Inactive Publication Date: 2010-07-14
SICHUAN AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Compared with the isocratic elution method, gradient elution has many disadvantages: for example, the operation is complicated, the requirement for the miscibility of the solvent as the mobile phase is high, and the requirement for the purity of the solvent is high; the mobile phase cannot be recycled, which increases the analysis time. Cost; having to re-equilibrate the column after each gradient run, time consuming, often causing baseline drift and loss of reproducibility, etc.
Using methanol as the mobile phase has two adverse effects. One is that the absorbance of methanol HPLC grade is larger than that of acetonitrile, and the noise generated during UV detection is large. Therefore, methanol HPLC grade is more sensitive than acetonitrile when performing high sensitivity analysis on UV short wavelength Poor (the analysis of fatty acids is in the short wavelength region); second, the viscosity of methanol increases after mixing with water, and excess pressure will be added to the column at the same flow rate, thereby reducing the service life of the chromatographic column

Method used

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  • Mixed fatty acid high performance liquid phase chromatography method and application
  • Mixed fatty acid high performance liquid phase chromatography method and application
  • Mixed fatty acid high performance liquid phase chromatography method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Mixed fatty acid analysis test.

[0028] 1. Instruments and main reagents

[0029] 1.1 Test equipment

[0030] Chromatographic instrument configuration: high performance liquid chromatography (Shimadzu (SHIMADZU) LC-20AD high performance liquid chromatography, Shimadzu CTO-10AS column thermostat, Shimadzu SPD-M20A diode array detector); record chromatograms (Shimadzu) Jin LC solution chromatography data workstation).

[0031] 1.2 Reagents

[0032] Standards: linolenic acid (C18:3), palmitoleic acid (C16:1), linoleic acid (C18:2), oleic acid (C18:1), palmitic acid (C16:0), and stearic acid ( C18:0) was purchased from Sigma Company of the United States, and the reference grade was obtained.

[0033] 2. Preparation of standard samples

[0034] 6 kinds of fatty acids were formulated into 1 mg / ml fatty acid solutions with methanol as solvent; equal amounts of fatty acid solutions were uniformly mixed to form fatty acid mixed solutions.

[0035] 3. Fatty acid lipidation...

Embodiment 2

[0046] Mixed fatty acid analysis test. It is no longer repeated in the same part as in Example 1, and the difference is that the chromatographic separation conditions are as follows:

[0047] Chromatographic column: Kromasil C18 (250mm×4.6mm, 5μm)

[0048] Column temperature: 10°C

[0049] Mobile phase: acetonitrile: 5% CTAB=90:10

[0050] Flow rate: 1.5ml / min

[0051] Detection wavelength: 242±5nm

[0052] The test chromatogram is shown in Figure 2.

Embodiment 3

[0054] Mixed fatty acid analysis test. It is no longer repeated in the same part as in Example 1, and the difference is that the chromatographic separation conditions are as follows:

[0055] Chromatographic column: Kromasil C18 (250mm×4.6mm, 5μm)

[0056] Column temperature: 15°C

[0057] Mobile phase: acetonitrile: water = 95:5

[0058] Flow rate: 1.5ml / min

[0059] Detection wavelength: 242±5nm.

[0060] The test chromatogram is as image 3 shown.

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Abstract

The invention discloses a mixed fatty acid high performance liquid phase chromatography method and application thereof. In order to solve the problems that the prior art has single method in separating linolenic acid, palmitic acid, linoleic acid and other three fatty acid mixtures with very similar chemical and physical properties with HPLC, and defects in elution method, mobile phase composition selection and the like, the invention provides a method for simultaneously separating six fatty acid mixtures, i.e. linolenic acid, palmitic acid, linoleic acid, oleic acid, palmitic acid and stearic acid with HPLC. The method esterifies fatty acid solution to be tested with the acetone solution of omega-bromoacetophenone and the acetone solution of triethanolamine, and separates the solution with HPLC. The HPLC adopts an isocratic elution method, and the mixture of acetonitrile and CTAB solution or acetonitrile-water solution is selected to serve as the mobile phase. Compared with the priorart, the invention has the advantages of simple and feasible operation, good separation effect, high detection sensitivity, accurate and reliable results, and simultaneous separation of 6 fatty acids.

Description

technical field [0001] The invention relates to a method for separating a mixture of fatty acids, in particular to a method for separating a mixture of fatty acids by high performance liquid chromatography and its application. Background technique [0002] Fatty acid is a long aliphatic hydrocarbon chain with a carboxyl group at one end. It is an important industrial raw material, an important component of the main nutrients of organisms and one of the main energy sources. With the improvement of people's living standards, the determination of fatty acids is becoming more and more important. In terms of food nutrition and safety, the determination of fatty acid composition and content of various edible oils, grains, fruits and milk powder is essential; in terms of medicine, the diagnosis of some diseases, the quality of Chinese herbal medicines, injections, etc. Relying on the analysis of fatty acid composition and content; in terms of energy, the evaluation and research of...

Claims

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Application Information

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IPC IPC(8): G01N30/50G01N30/06G01N30/36
Inventor 罗培高邓科君张勇胡学运陈俊伯任正隆
Owner SICHUAN AGRI UNIV
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