A body fluid pterin spectrum detection kit and its use

A kit and pterin spectrum technology are applied in chemical analysis and clinical medicine and medical fields, which can solve the problems of complex composition of pterin spectrum, difficult differential diagnosis, low sensitivity and specificity, etc.

Active Publication Date: 2021-10-15
SHANGHAI IND TECH INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the composition of the pterin spectrum in the urine sample is quite complicated. In addition to the above-mentioned NEO and BIO, there are also dihydroneopterin (7,8-dihydroneopterin, NH2), dihydrobiopterin (7,8-dihydrobiopterin, BH2), isoxanthopterin (isoxanthopterin, ISO), pterin (pterin, PT) and other pterin substances, and their relationship involves the action of multiple enzymes
Therefore, it is difficult to accurately differentiate all subtypes of BH4 deficiency based on the content of NEO and BIO in oxidized urine samples
On the other hand, the method based on HPLC-fluorescence detection is currently mainly used for the determination of NEO and BIO, but for other pterin substances in complex urine samples, the sensitivity and specificity are not high

Method used

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  • A body fluid pterin spectrum detection kit and its use
  • A body fluid pterin spectrum detection kit and its use
  • A body fluid pterin spectrum detection kit and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107] Optimization of urine oxidation conditions

[0108] In order to investigate the optimal oxidation time of pterin in urine under the condition of acidic manganese dioxide, a certain amount of urine sample was taken according to the operation steps and added with oxidizing agent and acidifying agent to react 0, 1, 2, 5, 10, 15, 20 respectively. , 25min, by comparing the mass spectrometric response peak areas of the four pterins NEO, BIO, ISO, and PT under different oxidation treatment times. the result shows( figure 1 ), the detection value of pterin reached a steady state after the oxidation time was 2 minutes. Considering the convenience of the experimental operation, the following oxidation time was determined to be 5 minutes.

Embodiment 2

[0110] Determination of mobile phase formulation

[0111] The four pterin standard solutions and actual urine samples were analyzed according to the formula in Table 3, and the corresponding chromatographic peak resolution, as well as the corresponding sensitivity and linear range were investigated, and the detection of actual urine samples could be completed. Preferably, a C18 reverse-phase chromatographic column is used, and when the mobile phase adopts formula 6 in Table 3, better sensitivity and a wider linear range are obtained.

[0112] Table 3 Optimization of mobile phase formulation

[0113]

Embodiment 3

[0115] A kind of tandem mass spectrometry pterin detection kit

[0116] 3.1 A tandem mass spectrometry pterin detection kit includes

[0117] Four pterin mixed standards: lyophilized powder, dry, 1 bottle, stored at -20°C;

[0118] A pterin standard product (internal standard): lyophilized powder, dry, 1 bottle, stored at -20°C;

[0119] Four pterin mixed standard quality control products (including urine matrix): lyophilized powder, dry, 3 concentrations, 3 bottles, stored at -20°C;

[0120] Manganese dioxide powder: dry, 1 bottle (2g);

[0121] Hydrochloric acid solution (1M HCl): 1 bottle (50mL);

[0122] Mobile phase: 2 bottles (each 1L), mobile phase A is acetonitrile (containing 1mM ammonium acetate and 20mM ammonia, 5% water), mobile phase B is water (containing 1mM ammonium acetate and 20mM ammonia, 5% acetonitrile);

[0123] U-bottom, heat-resistant microplate, 96 wells, 2 plates;

[0124] V-shaped cut-off bottom, clean microplate, 96-well detection plate, 2 plat...

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Abstract

The invention discloses a body fluid pterin spectrum detection kit and its use. The kit includes pterin mixed standards and creatinine.

Description

technical field [0001] The invention relates to the field of chemical analysis and clinical medicine and medicine. In particular, it relates to a new combination marker of pterin metabolites and its application in the preparation of a kit for screening hyperphenylalaninemia subtypes. Background technique [0002] Pterin compounds are derivatives of pteridine, which are widely distributed in blood, urine, cerebrospinal fluid and other body fluids in the human body, but the content is very small. Pterin compounds play an important auxiliary role in the process of cell metabolism. The content of pterin compounds in healthy human body fluid is low and stable. When the immune system is activated by certain diseases such as cancer (such as kidney cancer, breast cancer), the content of pterin compounds changes significantly. Hyperphenylalaninemia (HPA) is a common autosomal recessive metabolic disease that can lead to severe intellectual disability. Differences in genetic variat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 吴谦李园园李亦学贺珍
Owner SHANGHAI IND TECH INST
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