Surface plasma resonance (SPR) analysis method for determining combination level of quercetin and proteins

A surface plasma and analysis method technology, applied in the field of ion resonance analysis, can solve the problem of quercetin solubility, aggravate the solubility of quercetin and other problems

Inactive Publication Date: 2017-08-29
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the SPR analysis process often uses a buffer solution containing inorganic salts to prepare the sample solution to simulate the physiological environment in vivo, which further exacerbates the problem of low solubility of quercetin
At present, there is no report on the use of SPR method to study the interaction between quercetin and protein in the patent method, the reason is the solubility of quercetin in aqueous solution

Method used

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  • Surface plasma resonance (SPR) analysis method for determining combination level of quercetin and proteins
  • Surface plasma resonance (SPR) analysis method for determining combination level of quercetin and proteins
  • Surface plasma resonance (SPR) analysis method for determining combination level of quercetin and proteins

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Experimental program
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Embodiment

[0044] Embodiment: The analysis method of the present invention is used to determine the equilibrium constant of the interaction between quercetin and human serum albumin (HSA). Specific steps are as follows.

[0045] 1. Use a new sensor chip, which is a glass plate coated with a bare gold film. Install it into the SPR instrument, start the instrument, and maintain the temperature of the chip area at 37°C.

[0046] 2. Use 5mmol / L 3-mercaptopropionic acid aqueous solution to circulate for 30 minutes at a flow rate of 5 microliters / minute, and control the temperature of the chip area at 37°C. After completion, rinse the pipeline and chip surface with water.

[0047] 3. Dissolve HSA in acetic acid-sodium acetate buffer solutions with a sodium ion concentration of 10 mmol / L and a pH of 4.0, 4.5, 5.0, and 5.5, respectively, and circulate for 5 minutes at a flow rate of 5 μl / min. The pH of the solution with the largest signal change at the end of the circulation is 5.5, so the buf...

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Abstract

The invention discloses a surface plasma resonance (SPR) analysis method for determining a combination level of quercetin and proteins. An interaction system of the quercetin and the protines is used as an analysis target, and a balance constant of the interaction reaction, i.e. the combination level can be solved. In the method, the quercetin is dissolved by utilizing pyridine and diluted by utilizing a buffer solution containing the pyridine, so that the problem that the quercetin is insufficiently dissolved in an aqueous solution and the SPR analysis is difficult can be solved; and meanwhile, in the analysis process, when the dilute quercetin solution is prepared, the pyridine dilute solution of a same component without quercetin is correspondently prepared and used for representing the influence of the pyridine on an SPR signal so as to accurately acquire an interaction signal generated by the quercetin.

Description

technical field [0001] The invention relates to an application method of surface plasmon resonance technology in analytical chemistry, in particular to a surface plasmon resonance analysis method for measuring the interaction and binding level of quercetin and protein. Background technique [0002] Surface plasmon resonance, abbreviated as SPR (surface plasmon resonance), is an analytical method widely used in the analysis and detection of intermolecular interactions in substances, and can be used in the study of interactions between macromolecules, small molecules, and ions. The basic principle of SPR is to irradiate the SPR sensor chip with incident light under suitable conditions, and excite the evanescent wave generated by the incident light at the incident point to resonate with the surface plasmon wave of the thin metal layer on the surface of the sensor chip, so that the outgoing light After that, the interaction process occurs on the surface of the sensor chip, resul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552G01N1/38
CPCG01N21/553G01N1/38G01N2001/383
Inventor 刘虎威白玉张一丁
Owner PEKING UNIV
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