Method for detecting content of oxaloacetic acid on basis of high performance liquid chromatography

A technology of high performance liquid chromatography and oxaloacetic acid, which is applied in the field of oxaloacetic acid content determination based on high performance liquid chromatography (HPLC), can solve the problems of cumbersome sample pretreatment and high loss rate, and achieves obvious price advantages and high cost. Low, high detection sensitivity effect

Inactive Publication Date: 2018-10-12
SUZHOU COMIN BIOTECH
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Problems solved by technology

However, there are few reports on the detection technology of oxaloacetate based on high performance liquid chromatography on the market, and the pretreatment of samples is generally cumbersome and the loss rate is high

Method used

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Embodiment Construction

[0042] The present invention will be described in detail below in conjunction with embodiments.

[0043] A method for determining the content of oxaloacetate based on high performance liquid chromatography, the principle of which is that after the reaction of oxaloacetate and a derivative agent, a substance that absorbs under ultraviolet light is produced, separated by high performance liquid chromatography, and its content is measured by an ultraviolet detector ;

[0044] The method specifically includes the following steps:

[0045] Step 1 preparation of instruments and supplies;

[0046] Prepare high-performance liquid chromatography (water-resistant C18 column, UV detector), low-speed centrifuge, ultrasonic cleaner, solvent suction filtration device, 0.45 μm water filter membrane, 0.45 μm organic filter membrane, 50 0.22 μm water needle filters device, adjustable pipette, 50 2mL sample vials, methanol (chromatographic grade), potassium dihydrogen phosphate, dipotassium h...

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Abstract

The invention discloses a method for detecting the content of oxaloacetic acid on the basis of high performance liquid chromatography. According to a principle,after oxaloacetic acid reacts with a derivative agent to generate an ultraviolet absorbing substance,the ultraviolet absorbing substance is separated by means of the high performance liquid chromatography,and the content of the ultravioletabsorbing substance is detected through an ultraviolet detector. A high performance liquid chromatographic instrument needed to be used in the method has high precision,can detect the nanogram-scale content and has high detection sensitivity. A sample pretreatment process is simple and easy to operate. An extract has simple compositions,only a water solution is needed for extraction,and the cost is low. Detecting steps of the method are automatically operated through the instrument,hundreds of samples can be detected continuously,accurate data is read,and the time for detecting each sample isshort. Compared with a biological immunization detection method,the method has the advantages that the price advantage is obvious,and required reagents are basically free of toxicity.

Description

technical field [0001] The invention belongs to the field of life sciences, and in particular relates to a method for determining the content of oxaloacetate based on high performance liquid chromatography (HPLC). Background technique [0002] Oxaloacetate is an intermediate product of the tricarboxylic acid cycle, which is produced from malic acid under the catalysis of malate dehydrogenase. It condenses with acetyl-CoA to generate citric acid, starting a new cycle. Oxaloacetate can be converted from pyruvate and CO under the action of pyruvate carboxylase 2 Generated; it can also be generated from aspartic acid under the action of transaminase. In addition, oxaloacetate is also known as an inhibitor of succinate dehydrogenase. [0003] The existing detection methods of oxaloacetate in the market can be divided into two categories, including biological immunoassay technology and high performance liquid phase detection technology. The biological immunoassay method is susc...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/067
Inventor 王黔赵林川
Owner SUZHOU COMIN BIOTECH
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