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Fluorescent probe compound and its preparation method and its application and method for detecting endotoxin

A fluorescent probe and endotoxin technology, applied in the field of endotoxin detection, can solve the problems of fluorescence enhancement, fluorescent probe can only be used in glucose injection, HEPES buffer, ultrapure water or other non-containing, false positive, etc. , to achieve the effects of clear fluorescence curve, easy promotion and convenient operation

Active Publication Date: 2020-10-30
CHINA-SINGAPORE INT JOINT RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] Therefore, when the system to be tested contains components that can replace iodide ions in TPE-Be-I (for example, physiological saline and Ringer's solution), even without LPS, the fluorescence of the detection system will increase, thus causing false positives. As a result, the fluorescent probe can only be used for glucose injection, HEPES buffer, ultrapure water or other samples that do not contain iodide ion replacement components, therefore, there is still room for improvement

Method used

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  • Fluorescent probe compound and its preparation method and its application and method for detecting endotoxin
  • Fluorescent probe compound and its preparation method and its application and method for detecting endotoxin
  • Fluorescent probe compound and its preparation method and its application and method for detecting endotoxin

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Embodiment 1

[0069] A kind of fluorescent probe compound, fluorescent probe compound is (TPEPyE), molecular formula is C 33 h 28 NI, the structural formula is:

[0070]

[0071] The preparation method of fluorescent probe compound comprises the following steps:

[0072] S1. Add 3.7g (22mmol) diphenylmethane (1) into a 250mL two-necked flask; vacuumize the two-necked flask and then fill it with nitrogen, repeat three times; inject 40mL of anhydrous tetrahydrofuran (THF); The system was cooled to 0°C, and then 13.75mL (22mmol) n-butyllithium (1.6M) was added dropwise, and the reaction solution turned red; after stirring for 1 hour at 0°C (other embodiments can be made at 1°C, 2°C, 3 ℃, 4 ℃ under stirring for 1 hour), 5.222g (20mmol) 4-bromobenzophenone (2) was dissolved in 10mLTHF, and then slowly added dropwise to the reaction system; the reaction mixture was naturally warmed to room temperature and continued to react 10 hours. After the reaction is over, use saturated ammonium chlor...

Embodiment 2

[0081] In this embodiment, the solution to be tested is: a positive sample to be tested with a concentration of 0.9% sodium chloride injection with an endotoxin content of 0.75EU / mL and a concentration of 0.9% sodium chloride injection with an endotoxin content of 0.1EU / mL of negative test samples.

[0082] A kind of method utilizing the fluorescent probe compound detection concentration among the embodiment 1 to be the endotoxin content in the sodium chloride injection of 0.9%, comprises the following steps:

[0083] (1) Dissolve the fluorescent probe compound in pyrogen-free water to prepare a storage solution, specifically: take 1.1 mg of the fluorescent probe compound TPEPyE, pour it into a 25 mL volumetric flask, add pyrogen-free water to the volumetric flask to constant volume, Vibrate on a vortex shaker for 30 s to fully dissolve TPEPyE, and prepare a TPEPyE solution with a concentration of 100 μM, which is the storage solution.

[0084] (2) Preparation of standard solu...

Embodiment 3

[0102] In this embodiment, the solution to be tested is: a positive test sample with a concentration of 5% glucose injection with an endotoxin content of 0.75EU / mL and a negative sample with a concentration of 5% glucose injection with an endotoxin content of 0.1EU / mL. Sample to be tested.

[0103] A kind of method utilizing the fluorescent probe compound detection concentration among the embodiment 1 to be the endotoxin content in the glucose injection of 5%, comprises the following steps:

[0104] (1) Dissolve the fluorescent probe compound in pyrogen-free water to prepare a storage solution, specifically: take 1.65 mg of the fluorescent probe compound TPEPyE, pour it into a 25 mL volumetric flask, add pyrogen-free water to the volumetric flask to constant volume, Vibrate on a vortex shaker for 30 s to fully dissolve TPEPyE to prepare a TPEPyE solution with a concentration of 150 μM, which is the storage solution.

[0105] (2) Preparation of standard solution:

[0106] Tak...

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Abstract

The invention relates to the technical field of endotoxin detection, and provides a fluorescent probe compound to solve a problem that fluorescent probes only can be used for detecting samples free from iodide ion displacement components. The fluorescent probe compound is 4-(4-(1,2,2-triphenylvinyl)phenyl)ethylpyridinium iodide, the molecular formula of the compound is C33H28NI, and the structuralformula is shown in the description. A method for detecting endotoxin by using the fluorescent probe compound comprises the following steps: (1) preparing a stock solution; (2) preparing a standard substance solution; (3) preparing a standard reaction solution and a reaction solution to be detected; (4) detecting to obtain a standard fluorescence curve and a fluorescence curve of the reaction solution to be detected; and (5) comparing the fluorescence curve of the reaction solution to be detected with the standard fluorescence curve to determine whether the endotoxin content in the reaction solution to be detected reaches a standard or not. When a system to be detected contains a component capable of displacing iodide ions, TPEPyE has a good water solubility, and cannot form an aggregatein a solution free from LPS molecules, so fluorescence cannot be emitted, thereby the interference of iodide ion substitute to detection is eliminated.

Description

technical field [0001] The invention relates to the technical field of endotoxin detection, in particular to a fluorescent probe compound and its preparation method and its application and method for detecting endotoxin. Background technique [0002] Bacterial endotoxins widely exist in nature, for example, the endotoxin content in tap water is 1-100EU / mL. When endotoxin enters the human body through the digestive tract, it does not produce toxicity, but if endotoxin enters the blood through injection, etc., it will cause different diseases. After a small amount of endotoxin enters the blood, it can be inactivated by the Kupffer cells of the liver without causing damage to the body. A large amount of endotoxin entering the blood will cause a fever reaction, and even cause death in severe cases. Therefore, preparations such as biological products, injections, chemicals, radiopharmaceuticals, antibiotics, vaccines, dialysis fluids, and medical equipment (such as disposable s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D213/127C07D213/20C09K11/06G01N21/64
Inventor 唐本忠刘勇谢你姚必成
Owner CHINA-SINGAPORE INT JOINT RES INST
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