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Application of product for mass spectrometric detection of metabolite level in urine to preparation of product for early evaluation of intestinal polyp and colorectal cancer

A product, urine technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problem of lack of simultaneous detection, and achieve the effect of convenient, high sensitivity and high specificity for urine sampling

Pending Publication Date: 2022-01-11
广州济士源生物技术有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there is no simultaneous detection of N1, N12-diacetylspermine in urine (Hu Xin. The change of ornithine metabolic pathway in colon cancer and the application of multivariate data modeling in its diagnosis [D]. Jinzhou Medical University , 2021), kynurenine (Zhao Jianxing, Luo Xuemei, Zhan Yiming, Chen Bo, Zhu Dingliang. Simultaneous determination of kynurenine and kynurenic acid in human urine by high performance liquid chromatography-UV and fluorescence dual detectors[J]. Medicine, 2009,24(09):659-662.) and succinic acid (Yao Xunping, Li Xiaoping, Chen Xiaohong, Jin Micong. Study on the determination of binary organic acids in urine by high performance liquid chromatography fluorescence method[J]. Chinese Journal of Health Inspection ,2008(11):2190-2191+2196.) Detection technology of liquid chromatography tandem mass spectrometry

Method used

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  • Application of product for mass spectrometric detection of metabolite level in urine to preparation of product for early evaluation of intestinal polyp and colorectal cancer
  • Application of product for mass spectrometric detection of metabolite level in urine to preparation of product for early evaluation of intestinal polyp and colorectal cancer
  • Application of product for mass spectrometric detection of metabolite level in urine to preparation of product for early evaluation of intestinal polyp and colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1N1

[0041] Embodiment 1N1, the preparation of the standard working solution of N12-diacetylspermine, kynurene and succinic acid

[0042] 1. The source of the standard

[0043] N1, N12-Diacetylspermine standard product, the specification is 2.5mg, the purity is greater than 98%;

[0044] Kynurea standard product, the specification is 100mg, the purity is greater than 98%;

[0045] Succinic acid standard product, the specification is 250mg, the purity is greater than 98%.

[0046] The above standard products were purchased from Sigma Company.

[0047] 2. Configuration of N1, N12-diacetylspermine, kynurene and succinic acid standard working solution

[0048] Dilute each of the above standards to 10 gradients with ultrapure water.

[0049] N1, N12-diacetylspermine diluted concentrations were 0, 0.05, 0.4, 2, 4, 8, 16, 24, 32, 40ng / mL;

[0050] The concentration of kynurene after dilution is 0, 0.5, 4, 20, 40, 80, 160, 240, 320, 400ng / mL;

[0051] The concentrations of succinic a...

Embodiment 2

[0052] N1 in embodiment 2 standard substance aqueous solution, the mensuration of N12-diacetylspermine, kynurene and succinic acid content

[0053] Draw 10 μL of the calibration working solution with different concentrations prepared in Example 1, and detect by LC-MS / MS. The LC-MS / MS conditions are as follows.

[0054] (1) Chromatographic conditions:

[0055] Chromatographic column: Agilent Zorbax Eclipse XDB-C18 chromatographic column (100mm×2.1mm, 3.5μm);

[0056] Mobile phase reagent A: 10 mM ammonium formate aqueous solution (pH=3), mobile phase reagent B: 90% volume concentration acetonitrile solution of 10 mM ammonium formate (pH=3);

[0057] Column temperature 35°C;

[0058] Injection volume 10μL;

[0059] The mobile phase system selects reagent A with 95% volume concentration and reagent B with 5% volume concentration. The elution conditions select reagent A with 60% volume concentration and reagent B with 40% volume concentration for isocratic elution. The sample ...

Embodiment 3

[0067] N1 in embodiment 3 urine, the detection of N12-diacetylspermine, kynurene and succinic acid content

[0068] 1. Sample pretreatment

[0069] Take 1mL of urine sample, centrifuge at 12000r / min for 10min, pass the centrifuged mixture through a 0.22μm filter membrane, take the filtrate into a 10mL brown centrifuge tube, add 200μL of standard N1,N12-diacetylspermine (D6) , kynurene (ring-D4, 3, 4-D2) and succinic acid (D4) internal reference mixed solution for use, the concentrations were 10ng / mL, 200ng / mL and 400ng / mL respectively.

[0070] 2. Liquid chromatography tandem mass spectrometry (LC-MS / MS) analysis

[0071] The LC-MS / MS conditions of Example 2 were used for analysis. For the relevant chromatograms of N1, N12-diacetylspermine, kynurene and succinic acid Figure 1~3 . The results show that through the chromatograms of N1, N12-diacetylspermine, kynurene and succinic acid standard aqueous solution and urine, the chromatographic peaks of N1, N12-diacetylspermine,...

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Abstract

The invention discloses application of a product for mass spectrometric detection of a metabolite level in urine to preparation of a product for early evaluation of intestinal polyp and colorectal cancer, by using the product, a urine sample is subjected to impurity removal and then is directly introduced into a liquid chromatography tandem mass spectrometer for determination, and the content of N1, N12-diacetyl spermine, cynurin and succinic acid in urine can be rapidly and accurately detected. According to the method disclosed by the invention, corresponding standard substances of N1, N12-diacetyl spermine, cynurin and succinic acid are used as internal standard quantitative analytes, so that errors in a sample pretreatment process can be reduced; the selectivity and the accuracy of the method are improved by using liquid chromatography-tandem mass spectrometry; and the analysis time is shortened by selecting a chromatographic column and elution conditions. Meanwhile, an efficient and feasible technical method is provided for research and development of products for early diagnosis of adenomatous polyp and / or colorectal cancer.

Description

technical field [0001] The invention relates to the technical field of clinical testing and diagnosis, in particular to the application of a product for detecting metabolite levels in urine by mass spectrometry in the preparation of products for early evaluation of intestinal polyps and colorectal cancer. Background technique [0002] According to the World Health Organization, one-third of cancers can be prevented; one-third of cancers can be cured through early screening; one-third of cancers can use existing medical measures to prolong life, relieve pain and improve life quality. At present, cancer prevention and control can be carried out through tertiary prevention: primary prevention is etiological prevention, reducing the damage of external adverse factors; secondary prevention is early screening discovery, early diagnosis, and early treatment; tertiary prevention is improving quality of life , prolong the survival time. According to the "National Cancer Center Annu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/72G01N30/86
CPCG01N30/02G01N30/06G01N30/7266G01N30/8675G01N2030/045
Inventor 张娟张甜
Owner 广州济士源生物技术有限公司
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