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Method for detecting neopterin and biopterin in human urine

A technology for detection of human body and biopterin, applied in the field of detection, can solve the problems of non-compliance with rapidity, high throughput, poor storage stability, unfavorable promotion and use, etc., to achieve batch processing detection, low cost, and increase sample throughput Effect

Active Publication Date: 2019-12-06
GUANGZHOU KINGMED DIAGNOSTICS CENT
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Urine contains a large amount of water-soluble and highly polar impurities such as salt, and neopterin and biopterin are easily hydrolyzed and have poor storage stability. The 25cm C18 chromatographic column is used to enhance the retention and separation effect, and the analysis time is longer, which does not meet the requirements of rapid and high-throughput clinical medical testing.
At the same time, the detection cost of the traditional HILIC mode, LC-MS and other methods is relatively high, which is not conducive to popularization and use

Method used

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  • Method for detecting neopterin and biopterin in human urine
  • Method for detecting neopterin and biopterin in human urine
  • Method for detecting neopterin and biopterin in human urine

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Embodiment 1

[0059] 1. Collection and sample preservation of human urine

[0060] Three cases of fresh urine samples were collected, each sample was added with 6M HCl to adjust the pH to about 1, and then stored in the dark at -20°C and -70°C respectively. 15, 22 days each measured once.

[0061] 2. Pretreatment of urine samples

[0062] 1) Sampling: Leave to thaw to room temperature, mix well and take 420 μL of the acidified sample into a 1.5 mL centrifuge tube;

[0063] 2) Oxidation: Add 30 μL of 1% iodine / potassium iodide solution, shake for 3 minutes, and place in a centrifuge at 4°C for 30 minutes in the dark;

[0064] 3) Neutralization: short centrifugation for 3 seconds, sequentially add 5 μL 2mg / mL ascorbic acid, 15 μL 6M sodium hydroxide, vortex shaking for 3 minutes, 4°C, 12000rpm centrifugation for 5 minutes;

[0065] 4) Filtration: Use a 2.0 mL disposable syringe to pipette the supernatant through a 0.22 μm aqueous phase filter membrane for sample analysis.

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Abstract

The invention discloses an accurate, quick and high-throughput method for detecting neopterin and biopterin in the urine of a human body. The method comprises the following steps: analyzing a urine sample which is pre-treated by liquid chromatography on a chromatographic column which is a reverse bonded silica gel column; performing gradient elution with a mobile phase which is methanol and ammonium acetate buffer solution; and detecting with a fluorescent detector. The method is low in cost and simple in pre-treatment, can realize batch-processing detection to optimize the chromatographic process, shorten the analyzing time and greatly improve the throughput of sample, and is especially suitable for requirement of clinical and daily detection for the chromatographic process.

Description

technical field [0001] The invention relates to a detection method, in particular to a method for detection of neopterin and biopterin in human urine. Background technique [0002] Neopterin and biopterin are the metabolites of guanosine triphosphate (GTP) in the body, and guanosine triphosphate (GTP) is an important part in the synthesis of coenzyme tetrahydrobiopterin (tetrahy drobiopterin, BH4). Specopterin is mainly excreted through urine, and professionals can effectively distinguish the types of hyperphenylalaninemia through its content and the ratio of the two. Therefore, the differential diagnosis of early BH4 deficiency in children with HPA is very important. Atypical hyperphenylalaninemia and traditional hyperphenylalaninemia have different treatment options, and develop and establish a pretreatment method suitable for clinical medical testing, accurate, fast, and high-throughput chromatographic method It is beneficial to obtain accurate data quickly and in large...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 陈静宜程雅婷陈智刘伟霞赵蓓蓓李维佘旭辉禤学怡李卓阳董衡沙小花
Owner GUANGZHOU KINGMED DIAGNOSTICS CENT
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