Reagent and method for detecting homocysteine (HCY)

A technology for homocysteine ​​and reagents, which is applied in the field of homocysteine ​​detection and analysis, can solve the problems of time-consuming analysis procedures, high cost and the like, and achieve the effects of simple detection means, low cost and simple detection process.

Active Publication Date: 2013-01-23
STATE GRID CORP OF CHINA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these detection methods require time-consuming analytical procedures and expensive reagents and instruments

Method used

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  • Reagent and method for detecting homocysteine (HCY)
  • Reagent and method for detecting homocysteine (HCY)
  • Reagent and method for detecting homocysteine (HCY)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Synthesis of HMMPC: Mix 2mmoL of 3-hydroxymethyl-5-methylsalicylaldehyde, 3mmoL of cyclopentenone, 3mmoL of imidazole, 3mL of tetrahydrofuran and 3mL of deionized water, stir at room temperature for 48 hours, and dilute with 20mL of 1M HCl , and extracted three times with 30 mL of ethyl acetate, the organic phase was evaporated to dryness under reduced pressure, and then washed with ethyl acetate and petroleum ether in a volume ratio of 1:4 on a silica gel chromatography column to obtain HMMPC with a yield of 56%. The crystal was grown in ethyl acetate and petroleum ether to obtain a light yellow single crystal.

[0025] Characterization of HMMPCs:

[0026] 1 H NMR (600MHz, 25°C, DMSO- d6 ): δ7.18(s, 1H), 7.14(d, 1H), 7.01(s, 1H), 5.30(t, 1H), 2.58-2.63(m, 1H), 2.39-2.74(m, 2H), 2.11-2.38(m, 3H), 2.35(s, 3H); 13 C NMR (150MHz, CDCl 3 ): δ21.23, 28.92, 37.88, 61.94, 76.82, 122.48, 128.61, 129.21, 130.90, 132.36, 133.34, 151.60, 202.06; element analysis (calcd.%) C ...

Embodiment 2

[0028] Add 2ml pH7.0 HEPES (10mM) buffer solution and 2mM, 10μl HMMPC solution to three different UV cuvettes respectively, then add 70μl of Hcy, Cys, GSH respectively, and test on the UV-Vis spectrometer For detection, the UV-visible absorption diagram is shown in figure 2 .

Embodiment 3

[0030] Add 2ml pH7.0 HEPES (10mM) buffer solution and 2mM, 10μl HMMPC solution to different UV cuvettes respectively, add the same amount of various amino acids respectively, and measure the absorption value at 389nm in a UV-visible spectrometer. To draw a histogram of absorbance values ​​corresponding to different amino acids, see image 3 .

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Abstract

The invention provides a reagent for detecting homocysteine (HCY), which is a chromene derivate: 5-(hydroxylmethyl)-7-methyl-3, 3a-dihydrocyclopenta[b]chromen-1(2H)-one (HMMPC). The HMMPC reagent is prepared from 3-hydroxymethyl-5-methyl-salicylic aldehyde and 2-cyclopentenone in the next step at the room temperature; raw materials are cheap, the reaction condition is simple, and the scale production is easy. The invention provides a quantitative detection method for the homocysteine, which is used for quantitatively detecting the content of the homocysteine in a buffer solution of which the pH is 7.0 on account of HMMPC. In the detection method, high sensitivity and selectivity are shown on the HCY; the detection process is simple and convenient, sensitive and rapid; and the detection result is accurate.

Description

technical field [0001] The invention relates to homocysteine ​​detection and analysis technology, in particular to a reagent and method for rapid and quantitative detection of homocysteine. Background technique [0002] Cardiovascular and cerebrovascular diseases are a threat to the health of people all over the world. It is estimated that about 15 million people die of cardiovascular and cerebrovascular diseases in the world every year. Hyperhomocysteine ​​(Hcy)emia is an independent risk factor for cardiovascular and cerebrovascular diseases. At present, many laboratories have used Hcy as one of the clinical indicators that must be detected for the diagnosis of cardiovascular and cerebrovascular diseases. [0003] The currently reported detection method for detecting homocysteine ​​is: High Performance Liquid Chromatography (HPLC) (Kusmierek, K.; Glowacki, R.; Bald, E. Anal. Bioanal. Chem., 2006, 385, 855.), gas-mass chromatography (MacCoss, M.J.; Fukagawa, N.K.; Matthew...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D311/94
Inventor 霍方俊阴彩霞杨瑜涛张改清
Owner STATE GRID CORP OF CHINA
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