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Fluorescence detecting reagent, preparation method thereof and system and method for measuring skin cholesterol

A fluorescence detection and fluorescence measurement technology, used in the medical and optical fields, can solve the problems of difficult quality control, long detection time, and enzyme activity easily affected by environmental factors such as temperature and pH, so as to improve compliance and shorten detection time. Effect

Active Publication Date: 2018-06-26
ANHUI YIKANGDA PHOTOELECTRIC TECH CO LTD
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Problems solved by technology

This method involves two reagents, namely binding reagent and chromogenic reagent. The whole measurement process is about 4-5 minutes, and the detection time is relatively long; in addition, after the chromogenic reagent is added to the detection site, the color will gradually change with time. deepening, so the detection time of the chromogenic reagent needs to be strictly controlled, and the measurement results will vary greatly if it exceeds a few seconds, so the detection time needs to be very strict; moreover, the above-mentioned A-C-B reagents involve biological macromolecules (enzyme ), polymers and small molecules, the synthesis process is relatively complicated, and it is difficult to control the quality of the synthetic product in each step, and the enzyme activity is easily affected by environmental factors such as temperature and pH, and the storage time is relatively short

Method used

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  • Fluorescence detecting reagent, preparation method thereof and system and method for measuring skin cholesterol
  • Fluorescence detecting reagent, preparation method thereof and system and method for measuring skin cholesterol
  • Fluorescence detecting reagent, preparation method thereof and system and method for measuring skin cholesterol

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Experimental program
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Effect test

Embodiment 1

[0063] Such as figure 2 As shown, the synthesis of the fluorescence detection reagent 2-1 includes 7-diethylaminocoumarin-3-carboxylic acid 3-1 (261mg, 1mmol) and triethylene glycol 4-1 (1.5g, 10mmol) in EDCI [1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride] (288mg, 1.5mmol), DMAP (4-dimethylaminopyridine) (25mg, 0.2mmol ), Et 3 N(triethylamine) (152mg, 1.5mmol) and DCM (dichloromethane) (5mL) were reacted at room temperature (25°C) for 24h to obtain product 5-1 (338mg, 86%); (393mg, 1mmol) and succinic anhydride 6-1 (200mg, 2mmol) were refluxed for 24h in the presence of THF (tetrahydrofuran) (2mL) to obtain the product 7-1 (490mg, 99%); then the product 7-1 (493mg , 1mmol) and N-hydroxysuccinimide 8-1 (173mg, 1.5mmol) in EDCI [1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride] (288mg , 1.5mmol) and DCM (dichloromethane) (10mL) in the presence of the reaction to obtain the product 9-1 (550mg, 93%), the product 9-1 (12mg, 0.02mmol) and digitonin (12mg...

Embodiment 2

[0065] Such as image 3 As shown, the synthesis of the fluorescence detection reagent 2-2 includes the fluorescein 3-2 (332mg, 1mmol) and triethylene glycol 4-1 (1.5g, 10mmol) in EDCI [1-(3-dimethyl Aminopropyl)-3-ethylcarbodiimide hydrochloride] (288mg, 1.5mmol), DMAP (4-dimethylaminopyridine) (25mg, 0.2mmol), Et 3 N(triethylamine) (152mg, 1.5mmol) and DCM (dichloromethane) (5mL) were reacted at room temperature (25°C) for 24h to obtain product 5-2 (424mg, 88%); (482mg, 1mmol) and succinic anhydride 6-1 (200mg, 2mmol) were refluxed for 24h in the presence of THF (tetrahydrofuran) (2mL) to obtain the product 7-2 (580mg, 99%); then the product 7-2 (582mg , 1mmol) and N-hydroxysuccinimide 8-1 (173mg, 1.5mmol) in EDCI [1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride] (288mg , 1.5mmol) and DCM (dichloromethane) (10mL) in the presence of the reaction to obtain the product 9-2 (660mg, 94%), the product 9-2 (13mg, 0.02mmol) and digitonin (12mg, 0.01mmol ) in Et 3 N (t...

Embodiment 3

[0067] Such as Figure 4 As shown, the synthesis of the detection reagent 2-3 includes rhodamine 3-3 (444mg, 1mmol) and triethylene glycol 4-1 (1.5g, 10mmol) in EDCI [1-(3-dimethylamino Propyl)-3-ethylcarbodiimide hydrochloride] (288mg, 1.5mmol), DMAP (4-dimethylaminopyridine) (25mg, 0.2mmol), Et 3 N(triethylamine) (152mg, 1.5mmol) and DCM (dichloromethane) (5mL) were reacted to obtain product 5-3 (490mg, 85%) in the presence of conditions; then product 5-3 (576mg, 1mmol) and Succinic anhydride 6-1 (200mg, 2mmol) was refluxed in the presence of THF (tetrahydrofuran) (2mL) to obtain product 7-3 (672mg, 99%); then product 7-3 (676mg, 1mmol) and N-hydroxy Succinimide 8-1 (173mg, 1.5mmol) was dissolved in EDCI [1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride] (288mg, 1.5mmol) and DCM ( In the presence of dichloromethane) (10mL), the product 9-3 (735mg, 95%) was reacted, and the product 9-3 (16mg, 0.02mmol) and digitonin (12mg, 0.01mmol) were prepared in Et 3 N (trie...

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Abstract

The invention relates to a fluorescence detecting reagent, a preparation method thereof and a system and method for measuring skin cholesterol. The reagent, the preparation method and the system and method belong to the fields of medicine and optics. The digitonin capable of being combined with cholesterol is connected with different fluorophores in a chemical method. By means of the matched testing devices and testing materials, the detected fluorescence signal intensity can reflect the content of skin cholesterol, so as to assess the risk of skin cholesterol-related diseases like cardiovascular disease and achieve the effect of early warning and prevention to guide people to reasonably plan the diet and exercise habits, so that the body is always in a healthy state. The high-sensitivityfluorescence detecting reagent is adopted to overcome the shortcomings that the skin cholesterol is low in content and difficult to detect and achieve non-invasive rapid sensitive detection of skin cholesterol.

Description

technical field [0001] The invention belongs to the fields of medicine and optics, and particularly relates to the application field of detection reagents in the prediction and prevention of chronic diseases, in particular to a fluorescence detection reagent and its preparation method, and a measurement system and method for skin cholesterol. Background technique [0002] With the aggravation of population aging and changes in eating habits, the prevalence of atherosclerotic diseases is gradually increasing. Such diseases belong to the category of chronic diseases. Seriously affect the quality of life of patients, in view of this, it is urgent to develop a marker and detection method that can early warning of such diseases. [0003] Studies have shown that hypercholesterolemia has a great correlation with atherosclerosis. The determination of plasma total cholesterol, high-density lipoprotein (HDL) and low-density lipoprotein (LDL) content is one of the widely used methods ...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/64G01N2021/6417
Inventor 刘勇王贻坤倪敬书董美丽
Owner ANHUI YIKANGDA PHOTOELECTRIC TECH CO LTD
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