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Detection method for o-dihydroxy metabolite spectrum in urine

A detection method, a technique of ortho-2, applied in the field of metabolome analysis

Inactive Publication Date: 2009-05-13
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, there is no report at home and abroad on the use of RHPLC-MS-based metabolomics technology to obtain the urine-containing metabolite profile of breast cancer patients

Method used

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  • Detection method for o-dihydroxy metabolite spectrum in urine
  • Detection method for o-dihydroxy metabolite spectrum in urine
  • Detection method for o-dihydroxy metabolite spectrum in urine

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0017] Example 1 Using fast liquid chromatography-mass spectrometry to analyze the metabolites of ortho-dihydroxyl compounds in normal human urine

[0018] Sample pretreatment: Take 3mL solid phase extraction empty column and fill 100mg phenylboronic acid filler. The pretreatment procedure is: 15mL 50% methanol water rinse containing 0.1mol / L formic acid, 15mL 50% methanol rinse, 20mL 0.25mol / L ammonium acetate (pH8.6) Rinse. After the urine sample is thawed at room temperature, mix and centrifuge. Accurately measure 0.8mL supernatant and add 0.06mL (221.8μg / mL) internal standard (catechin) solution, mix well, and load the sample on the pretreated phenylboronic acid column. , Add 0.2mL 0.25mol / L ammonium acetate (pH8.6) and let stand for 5min. Then wash with 1.6mL 0.25mol / L ammonium acetate (pH8.6), 0.25mL 50% methanol, and 0.2mL 50% methanol water containing 0.1mol / L formic acid, and finally containing o-dihydroxy metabolites with 2mL containing 0.1 mol / L formic acid is eluted wi...

Embodiment 2

[0023] Basically the same as Example 1, the difference is that the analysis object is the urine of breast cancer patients, and the analysis results are shown in the attachment figure 2 , Is a total ion chromatogram (TIC) of o-dihydroxy metabolites in the urine of breast cancer patients. 170 o-dihydroxy metabolites were detected in total.

Embodiment 3

[0025] Basically the same as Example 1, the difference is that the analysis object is the urine of breast cancer patients after chemotherapy. The analysis results are shown in the appendix image 3 , Is the total ion current diagram (TIC) of ortho-dihydroxy metabolites in the urine of breast cancer patients after chemotherapy. A total of 171 ortho-dihydroxy metabolites were detected.

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Abstract

The invention provides a method for testing a spectrum of o-dihydroxy metabolites in urine. Solid-phase extraction is carried out by using phenylo boric acid so as to purify and enrich the o-dihydroxy metabolites; and liquid chromatography-mass spectrometry is adopted to test a characteristic spectrum of the o-dihydroxy metabolites. Concentration by centrifuging is carried out by collecting formic acid-methanol water eluate; and residua are dissolved in water and analyzed after centrifugation. The invention provides a new method for testing the spectrum of the o-dihydroxy metabolites and lays a foundation for further discovering new diagnostic markers of breast cancer.

Description

Technical field [0001] The invention belongs to the technical field of metabolome analysis, and particularly relates to a detection method for the spectrum of metabolites containing o-dihydroxyl groups in urine. Background technique [0002] Breast cancer is a malignant tumor that seriously endangers women's health. Its incidence has always ranked first among female tumors in the world, accounting for 1 / 3 of female malignancies, and it is still increasing at a rate of 2% every year. So far, the exact cause of breast cancer is not very clear, and its recurrence and metastasis rate is very high. Early diagnosis and early treatment are the key to increasing the survival rate of patients and significantly improving the prognosis and quality of life of patients. Although imaging examinations such as B-ultrasound, CT and MRI are more reliable methods for diagnosis and prognosis, the radiation used in CT and MRI can damage the human body to a certain extent. B-ultrasound is poor in clar...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06G01N33/48
Inventor 王书芳沈朋邵青刘昌孝程翼宇
Owner ZHEJIANG UNIV
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