Mobile phase for separating levodopa in broad beans in high efficiency liquid chromatography

A high-performance liquid chromatography and levodopa technology, which is applied in the mobile phase field of separating levodopa in broad beans in high-performance liquid chromatography, can solve problems such as seal ring friction, damage to the pump system, damage, etc., to reduce damage, The effect of increasing the pH value and facilitating quantitative analysis

Inactive Publication Date: 2012-09-19
CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High-performance liquid chromatography does not have high requirements on the absorption wavelength of the mobile phase, but the mobile phase with corrosion and precipitation will cause damage to the components of the high-performance liquid chromatography. For example, the phosphate buffer in the existing mobile phase is prone to precipitation, causing Piston rod and seal rubbing, damaging the pump system

Method used

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  • Mobile phase for separating levodopa in broad beans in high efficiency liquid chromatography
  • Mobile phase for separating levodopa in broad beans in high efficiency liquid chromatography
  • Mobile phase for separating levodopa in broad beans in high efficiency liquid chromatography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1, such as figure 1 Or as shown in 2, the mobile phase is composed of 95% by volume of 0.1% formic acid and 5% acetonitrile, and the HPLC-UV chromatograms obtained by separating and detecting the levodopa standard and the levodopa extracted from Vicia faba flower respectively . Chromatographic column is Shiseido CAPCELL PAK CR column (150 mm×4.6 mm, 5 μm); mobile phase flow rate: 1.0 mL / min; detection wavelength: 280 nm; column temperature: 30 ℃; injection volume: 20 μL, in the chromatogram The abscissa is the retention time of the levodopa standard and the levodopa extracted from the broad bean flower passing through the chromatographic column, and the time is in minutes, and the ordinate is the levodopa standard and the levodopa extracted from the broad bean flower passing through the chromatographic column Response value voltage, the voltage response value is in volts (the following examples use the same chromatographic column conditions as this example). f...

Embodiment 2

[0036] Example 2, such as image 3Or as shown in 4, adopt mobile phase composition volume percent to be: 0.1% formic acid 90%, acetonitrile 9%, 0.1 mol / L acetic acid 1%, the levodopa standard substance and the levodopa extracted in the broad bean flower are separated respectively Detect the resulting HPLC-UV chromatogram, figure 1 As shown in the chromatogram, the mobile phase can completely separate the baseline of levodopa, and the peak area of ​​the peak 4.465 has a good linear correlation with the mass, but the baseline drift is obvious. figure 2 As shown in the chromatogram, the mobile phase can completely separate the baseline of levodopa and some unknown substances, and the peak area of ​​the peak 4.490 has a good linear correlation with the mass, but the baseline drift is obvious.

Embodiment 3

[0037] Example 3, such as Figure 5 Or as shown in 6, adopt mobile phase composition volume percentage to be: 0.1% formic acid 97%, acetonitrile 2%, 0.1 mol / L acetic acid 1%, the levodopa standard substance and the levodopa extracted in the broad bean flower are separated respectively According to the HPLC-UV chromatogram obtained by detection, the separation of levodopa between the standard levodopa and the faba bean flower sample is better in this mobile phase, and the retention time reaches more than 6 minutes, but the baseline drift of the two is obvious.

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Abstract

The invention discloses a mobile phase for separating levodopa in broad beans in high efficiency liquid chromatography. The mobile phase contains, by volume percent: 69-97% of 0.1% formic acid, 2-30% of acetonitrile and 1-10% of 0.1mol / L acetic acid. Sediment is prevented from being generated by using acetic acid to replace phosphate buffer fluid, and damage to a pump system is reduced. The mobile phase has no damage to the pump system of a high efficiency liquid chromatograph after being used for a long time. The formic acid can be completely dissolved in acetonitrile-formic acid-water with any proportion at any proportion, no sediment is generated, and the pump system cannot be damaged and is good in condition in year-and-year use. When used for isocratic elution, the mobile phase is simple to operate, time-saving, good in repeatability and easy to repeat.

Description

technical field [0001] The invention relates to a method for determining the content of levodopa components in organisms, in particular to a mobile phase for separating levodopa in broad beans in high performance liquid chromatography. Background technique [0002] Levodopa (L-DOPA), namely 3,4-dihydroxyphenylalanine, is the metabolic precursor of catecholamine hormones, neurotransmitters (such as norepinephrine, dopamine) and melanin in organisms. It is used to treat Parkinson's disease (Parkinson's disease), so it is of great significance to measure the content of levodopa in the pharmaceutical preparation and biological samples. In recent years, the in vivo and in vitro analysis methods of levodopa have made great progress. Ultraviolet spectrophotometry and high performance liquid chromatography are the most widely used in the in vivo and in vitro analysis of levodopa. Flow injection automatic analysis method and capillary electrophoresis, etc. It is also widely used, an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/26
Inventor 郑开斌李爱萍郑金贵廖素凤曹奕鸯
Owner CROP RES INST OF FUJIAN ACAD OF AGRI SCI
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