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Fluorescence ratio probe as well as synthesis method thereof and application thereof to detection on intracellular lysosomal formaldehyde

A technology of fluorescent ratio probes and synthesis methods, which is applied in the directions of fluorescence/phosphorescence, chemical instruments and methods, and luminescent materials. Targeting, good water solubility, and high sensitivity

Active Publication Date: 2019-01-04
NORTHEASTERN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the complex synthesis process, fast photobleaching, short fluorescence lifetime, poor water solubility, detection accuracy easily interfered by the probe environment, and low detection sensitivity faced by the existing fluorescent probes when detecting formaldehyde, the present invention adopts water-soluble Combining nano-carbon dots with good properties and small organic molecules, a ratiometric formaldehyde fluorescent probe with dual emission bands is synthesized, abbreviated as CDs-NA; and further provides the synthesis method and application of CDs-NA

Method used

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  • Fluorescence ratio probe as well as synthesis method thereof and application thereof to detection on intracellular lysosomal formaldehyde
  • Fluorescence ratio probe as well as synthesis method thereof and application thereof to detection on intracellular lysosomal formaldehyde
  • Fluorescence ratio probe as well as synthesis method thereof and application thereof to detection on intracellular lysosomal formaldehyde

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1: the synthetic method of compound CDs-NA comprises the following steps:

[0058] Step 1: Preparation of methionine carbon dot solid powder

[0059] Weigh 0.8g of methionine and 1g of citric acid into a 20mL small beaker, slowly add 5mL of secondary water and stir evenly with a glass rod to dissolve them. After the dissolution is complete, the reaction solution is obtained;

[0060] All the reaction solution was transferred to a 25mL reaction kettle, and a steel jacket was installed, and the reaction was heated at 200°C for 3h in a forced air drying oven. After the reaction was completed, it was cooled to room temperature to obtain a mixed solution.

[0061] Next, the mixture was aspirated and centrifuged at a speed of 12000r / min for 15 minutes to remove non-fluorescent large particles and obtain a supernatant of carbon dots for future use.

[0062] Then, the supernatant was dialyzed with a dialysis bag with a molecular weight cut-off of 500D for 12 hours t...

Embodiment 2

[0074] Embodiment 2: Fluorescence ratio probe CDs-NA changes with the addition of fluorescence spectrogram of different amounts of formaldehyde

[0075] Use PBS (10mM, pH 7.4) as a solvent, and prepare a concentration of 0.1mg mL -1 CDs-NA in PBS solution. Add 100 μL of formaldehyde solutions of different concentrations (0, 1, 2, 5, 10, 15, 20, 30, 40, 50, 60, 100, 160 μM) to the centrifuge tube containing 400 μL of CDs-Na in PBS solution, and shake for 30 minutes . Using a cuvette with an optical path of 0.5 cm, a fixed excitation wavelength of 365 nm, and a photomultiplier tube voltage of 650 V, measure its fluorescence emission spectrum with a Hitachi F-7000 fluorescence spectrophotometer, and the probe CDs-NA of different amounts of formaldehyde For changes in the fluorescence spectrum, see figure 2 ,From figure 2 It can be seen that with the increase of formaldehyde concentration, the fluorescence peak at 414nm remains unchanged, while the fluorescence peak at 535nm...

Embodiment 3

[0076] Embodiment 3: the calculation of probe CDs-NA detection formaldehyde lower limit

[0077] According to the fluorescence spectrogram of example 2, the fluorescence intensity ratio (F) of the fluorescence intensity at the 535nm place and the 414nm place 535 / F 414 ) and the corresponding formaldehyde concentration, see image 3 . Depend on image 3 Visible, formaldehyde concentration is 0-40μmol / L, F 535 / F 414 Good linearity with formaldehyde concentration. According to the formula Dl=3σ / k (Dl is the lower limit of detection, σ is the standard deviation of the intercept, and k is the slope), the lower limit of detection can be calculated as 338nM.

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Abstract

The invention relates to a fluorescence ratio probe as well as a synthesis method thereof and application thereof to detection on intracellular lysosomal formaldehyde, and belongs to the field of analytic chemistry. The structural formula of the fluorescence ratio probe is shown in the description. The preparation method comprises the following steps: preparing methionine carbon dot solid powder;and preparing a fluorescence ratio probe crude product and substituting by using hydrazine hydrate to obtain the fluorescence ratio probe, wherein the fluorescence ratio probe is used for detection formaldehyde and particularly used for detecting formaldehyde in the water environment and formaldehyde in a biological sample. The fluorescence ratio probe successfully realizes detection of intracellular and extracellular formaldehyde, has high selectivity and detection sensitivity and provides important theoretical significance in that the double-emission fluorescence ratio probe is further applied to the fields of biomarkers and the like in the future.

Description

technical field [0001] The invention belongs to the technical field of analytical chemistry and relates to a fluorescent ratio probe for detecting intracellular formaldehyde; the invention also relates to a synthesis method of the fluorescence ratio probe; in addition, the invention also relates to the application of the fluorescence ratio probe. It specifically relates to a fluorescent ratio probe, its synthesis method and the application of detecting intracellular lysosome formaldehyde. Background technique [0002] Formaldehyde (FA) is a common carcinogen and teratogen, and has been identified as a potential allergen by the World Health Organization. As a chemical pollutant, indoor air, atmospheric environment and food pollution caused by formaldehyde can lead to formaldehyde poisoning, causing dizziness, headache or nausea, vomiting, severe memory loss, and even death. However, it has been reported that FA exists in cells at relatively high concentrations under normal p...

Claims

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Application Information

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IPC IPC(8): C07D221/14C09K11/06G01N21/64
CPCC07D221/14C09K11/06C09K2211/1029G01N21/643
Inventor 陈帅贾叶于永亮
Owner NORTHEASTERN UNIV
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