Preparation method for nano chemical light emitting probe capable of natural degrading in living body

A technology of chemiluminescence and chemiluminescence reagents, which is applied in the field of preparation of nanoscale chemiluminescence probes, can solve the problems of unobservable reaction phenomena and difficult real-time detection, etc., and achieves simple production methods, improved detection sensitivity, and obvious experimental results Effect

Inactive Publication Date: 2005-10-19
SOUTH CHINA NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Chemiluminescent probes are mainly used to detect free radicals, and the reaction between the two is instantaneous and violent. Therefore, it is difficult for the existing chemiluminescent probes to detect the chemiluminescence of biological tissues in vivo in real time. As a result, some reaction phenomena cannot be observed, resulting in experimental artifacts

Method used

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  • Preparation method for nano chemical light emitting probe capable of natural degrading in living body
  • Preparation method for nano chemical light emitting probe capable of natural degrading in living body
  • Preparation method for nano chemical light emitting probe capable of natural degrading in living body

Examples

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Effect test

Embodiment 1

[0024] (1) Human serum albumin 10mg, chemiluminescence probe MCLA concentration 20umol / L, rapeseed oil 4ml, triple distilled water 2ml, nonionic surfactant Tween 80 concentration 1%.

[0025] (2) Under the condition of avoiding light, mix the human serum albumin solution with the chemiluminescence reagent MCLA solution diluted to the working concentration, then add rapeseed oil to it, and treat it with 150W ultrasonic wave for 1min at 0°C ; After adding the non-ionic surfactant Tween 80 with a concentration of 1%, continue to use ultrasonic treatment for 2 minutes, the mixture presents a uniform milky white.

[0026] (3) Divide the prepared emulsion, and centrifuge at 2000g / min for 10min at 4°C. After centrifugation, the oil from the emulsion was gently removed. This step is repeated several times until the oil separated out by centrifugation is basically removed.

[0027] (4) Heat and solidify the white emulsion obtained after centrifugation in an environment of 80° C. for ...

Embodiment 2

[0029] Human serum albumin 5mg, chemiluminescent probe MCLA concentration 10umol / L, rapeseed oil 2ml, triple distilled water 1ml, mix evenly; under the condition of 0℃, 80W ultrasonic treatment for 2min, add 0.2% non-ionic After surfactant Tween 80, continue to use ultrasonic treatment for 3 minutes. The prepared emulsion was subpackaged, centrifuged, and excess oil was absorbed, heated and solidified at 50° C. for 20 minutes, and then stored in a refrigerator at 4° C. in the dark.

Embodiment 3

[0031] Bovine serum albumin 50mg, chemiluminescence probe MCLA concentration 20umol / L, rapeseed oil 4ml, three-distilled water 2ml, mix evenly; under the condition of 0℃, 220W ultrasonic treatment for 1min, add 0.2% non-ionic After surfactant Tween 80, continue to use ultrasonic treatment for 1min. The prepared emulsion was divided into packages, centrifuged, excess oil was sucked off, heated and solidified at 80° C. for 10 min, and then stored in a refrigerator at 4° C. in the dark.

[0032] Wherein, the raw material that embodiment 1 adopts, consumption and condition are best among 3 examples.

[0033] as the picture shows, figure 1 It is a photo taken when the nanoscale chemiluminescent probe made according to the method proposed in the present invention was observed with a Japanese H-300 transmission electron microscope. For the results corresponding to Example 1, the particle size is 50-150 nm. Magnification 50k.

[0034] figure 2 In the in vitro experiment, the det...

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Abstract

The characteristics of the chemiluminescnece probe are that the soluble protein with the grain sizes less than 150nm coats the surface of the chemiluminescence reagent. The preparing method includes following steps. (1) chemiluminescence reagent, with the vegetable oil being added into. Under 0-4 deg.C, the amixture is processed by the ultrasonic in the intensity within 80-220W, for 2-5 min, with the nonionic surfactant being added into. The admixture presents the even ivory. (2) The emulsion produced is sub-packaged. The precipitable oil caused by centrifugalizing the emulsion is removed. (3) Heating solidifies the emulsion. The solidified emulsion is conserved at the temperature lower than 4 deg.C. The invention solves the instantaneous reaction between the chemiluminescence reagent MCLA and the oxygen-derived free radicals.

Description

technical field [0001] The invention relates to a preparation method of a nanoscale chemiluminescent probe that can be naturally degraded in a living body. Background technique [0002] Chemiluminescent probes are mainly used to detect free radicals, and the reaction between the two is instantaneous and violent. Therefore, it is difficult for the existing chemiluminescent probes to detect the chemiluminescence of biological tissues in vivo in real time. As a result, some reaction phenomena cannot be observed, resulting in experimental artifacts. Contents of the invention [0003] The object of the present invention is to provide a kind of preparation method of the nano-scale chemiluminescent probe that can degrade naturally in organisms, solve the problem of simple chemiluminescent reagent MCLA and oxygen free radical ( 1 o 2 , O 2 - ) is instantaneous and short-lived, it is difficult to achieve real-time long-term observation. [0004] The nano-scale chemiluminescenc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76
Inventor 邢达郝敏
Owner SOUTH CHINA NORMAL UNIVERSITY
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