Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with two-step derivation method

A technology for simultaneous analysis and carbohydrate metabolism, applied in the field of simultaneous analysis of two-step derivatization of bio-based samples

Active Publication Date: 2012-08-01
王益超
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] Aiming at the defects of existing analysis methods such as chromatographic column ion exchange method, organic acid extraction method and urease pretreatment method, the purpose

Method used

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  • Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with two-step derivation method
  • Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with two-step derivation method
  • Method for synchronously analyzing base, nucleotide, organic acid, fatty acid, amino acid and saccharide metabolic product with two-step derivation method

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Embodiment 1

[0036] 1) Sampling: Use test tubes or filter paper to collect urine: use ordinary test tubes or domestic No. 3 filter paper, cut into 3.0cmX8.0cm size. The morning urine or any time of urine soaked, filtered and dried, put it in a special filter paper drying bag for later use.

[0037] 2) Collect test samples: dry the filter paper bio-matrix sample, elute with 1.0mL deionized double-distilled water several times, and collect the eluate after the bio-based sample is processed by physical methods such as ultrasound and centrifugation; measure the creatinine in the eluate content, to determine the creatinine concentration, take 200 μl of eluate equivalent to 2.50 μmoL / μl creatinine concentration as the test sample for later use;

[0038] 3) Oximation treatment: Add 50 μl of hydroxylamine hydrochloride aqueous solution (containing 25 μg / μl of hydroxylamine hydrochloride) to 200 μl of the test sample, adjust the pH value of the above test sample to 10 with 6 normal concentration Na...

Embodiment 2

[0059] The preparation method and analysis method are the same as in Example 1, except that the bio-based sample is changed to blood, and the same detection effect as in Example 1 can be obtained, see figure 2 .

[0060] The present invention can also detect other bio-based samples, such as cerebrospinal fluid, pathological tissue fluid and amniotic fluid, etc., and can obtain the same detection effect as in Example 1, which will not be listed here.

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Abstract

The invention discloses a method for synchronously analyzing a base, a nucleotide, an organic acid, a fatty acid, an amino acid and a saccharide metabolic product with a two-step derivation method. The method comprises the following steps of: performing a series of physicochemical method technical treatment such as ultrasonic treatment, centrifugation, oximation, urea removing, phosphorus removing, sulfur removing, protein removing, nitrogen blowing, vacuum drying, and trimethyl silylation derivation on biological substrate samples (such as urea, blood, cerebrospinal fluid, and tissue fluid); and detecting the biological substrate samples by adopting a gas chromatograph-mass spectrum combination technology. Due to the adoption of method, programmed treatment can be performed on the biological substrate samples simultaneously, and over 400 kinds of metabolic intermediate and final products of over five kinds of substances can be detected at one time.

Description

technical field [0001] The invention belongs to the field of synchronous analysis methods of two-step derivatization method for bio-based samples, in particular to the simultaneous analysis of bases, nucleosides, organic acids, fatty acids, amino acids and Methods of Carbohydrate Metabolites. Background technique [0002] Human urine contains terminal or intermediate metabolites of many endogenous compounds such as bases, nucleosides, organic acids, fatty acids, amino acids, and sugars. These organic compounds have relatively high water solubility and renal clearance rate. The concentration is usually higher than serum and other tissue fluids, such as urine, blood, CSF (cerebrospinal fluid) and interstitial fluid, among which urine specimens are the easiest to collect and are non-invasive techniques recognized by researchers at home and abroad. In the 20th century, our country can only carry out preliminary detection of genetic metabolic, nutritional and other metabolic dis...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 王益超
Owner 王益超
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