In the field of chemiluminescence immunoassay, acridine esters have become important chemiluminescence markers due to their high sensitivity and low background signal advantages. However, the chemiluminescence reaction of acridine ester requires specific reaction conditions, in which the pre excitation solution and excitation solution are key components for initiating this reaction. This article will systematically explain the mechanism of action, preparation requirements, and usage precautions of acridine ester pre excitation solution and excitation solution, helping experimental personnel to gain a deeper understanding of this important chemical reaction system.

acridine ester powder

Pre excitation solution: mechanism of action of hydrogen peroxide solution

The pre excitation solution is the first step in the chemiluminescence reaction of acridine ester, and its main component is hydrogen peroxide solution. Hydrogen peroxide is relatively stable under acidic conditions and can undergo preliminary reactions with the active groups in acridine ester molecules. In the pre excitation stage, hydrogen peroxide attacks the active ester groups of acridine ester molecules, forming unstable peroxide intermediates. This reaction is usually carried out under acidic or neutral conditions to ensure the stability and reaction efficiency of hydrogen peroxide.

Stimulating solution: the key role of sodium hydroxide solution

The excitation solution is the second step of the acridine ester chemiluminescence reaction, and its main component is sodium hydroxide solution. After the pre excitation solution reacts with acridine ester to form a peroxide intermediate, an alkaline excitation solution is added to rapidly increase the pH value of the reaction system. Under strong alkaline conditions, peroxide intermediates undergo decomposition, producing excited state acridone molecules. When the excited state molecules return to the ground state, photons are released, producing a chemiluminescence signal.

The synergistic effect of pre excitation liquid and excitation liquid

Pre excitation solution and excitation solution play a synergistic role in the chemiluminescence reaction of acridine ester. Hydrogen peroxide in the pre excitation solution reacts with acridine ester under acidic conditions to form an unstable peroxide intermediate; The sodium hydroxide in the excitation solution provides a strong alkaline environment, promoting the decomposition of peroxide intermediates and generating chemiluminescence signals.

Preparation requirements for pre excitation solution and excitation solution

When preparing pre excitation solution and excitation solution, strict quality control standards must be followed. Firstly, all raw materials must use high-purity reagents to avoid the introduction of impurities such as metal ions and carbonates; Secondly, prepare the solution with ultrapure water to avoid ion interference; Thirdly, the prepared solution needs to be filtered and sterilized, and stored at 4 ℃ in the dark to avoid degradation of the reagent due to light and high temperature; Fourth, regularly check the pH value and concentration of the solution to ensure that the quality of the reagents meets the requirements; Fifth, establish a record of reagent usage, including preparation date, batch number, and usage status, for easy traceability and quality control.

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