Method for detecting alkaline phosphatase activity by molybdenum disulfide quantum dot inner-filter effect fluorescence

A technology of molybdenum disulfide and quantum dots, which is applied in the field of nanomaterials, can solve problems such as less research on optical properties, and achieve the effect of reducing preparation costs and efficient detection

Active Publication Date: 2018-08-17
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, compared with its electrical and catalytic properties, MoS 2 The optica...

Method used

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  • Method for detecting alkaline phosphatase activity by molybdenum disulfide quantum dot inner-filter effect fluorescence
  • Method for detecting alkaline phosphatase activity by molybdenum disulfide quantum dot inner-filter effect fluorescence
  • Method for detecting alkaline phosphatase activity by molybdenum disulfide quantum dot inner-filter effect fluorescence

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Experimental program
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Effect test

Embodiment 1

[0038]Embodiment 1, preparation molybdenum disulfide quantum dot

[0039] Dissolve 100 mg of molybdenum disulfide powder and 0.6 g of sodium hydroxide in 11 mL of 85% ethanol / water, sonicate at 580 W for 20 h, let stand at room temperature for 12 h, collect the supernatant, add concentrated hydrochloric acid to adjust the pH of the solution After reaching neutrality, filter to remove insoluble matter, dialyze with a dialysis bag of 1000 KDa for 24 h, then filter through a 0.22 µm microporous membrane, vacuum freeze-dry, and redisperse the obtained solid powder into water to obtain molybdenum disulfide quantum dot solution . like figure 1 and figure 2 As shown, the synthesized molybdenum disulfide quantum dots have tunable luminescence properties, and they can fluoresce under the excitation of 360 nm to 500 nm light, and their maximum excitation and emission wavelengths are 400 nm and 500 nm, respectively. image 3 is the UV spectrum of the synthesized molybdenum disulfide ...

Embodiment 2

[0040] Embodiment 2, prepare molybdenum disulfide quantum dot

[0041] Dissolve 200 mg of molybdenum disulfide powder and 0.8 g of sodium hydroxide in 11 mL of 85% ethanol / water, sonicate at 580 W for 12 h, let stand at room temperature for 12 h, collect the supernatant, add concentrated sulfuric acid to adjust the pH of the solution After reaching neutrality, filter to remove insoluble matter, dialyze with a dialysis bag of 1000 KDa for 24 h, then filter through a 0.22 µm microporous membrane, vacuum freeze-dry, and redisperse the obtained solid powder into water to obtain molybdenum disulfide quantum dot solution .

Embodiment 3

[0042] Embodiment 3, the detection of alkaline phosphatase

[0043] In order to further study the feasibility of detecting alkaline phosphatase by the inner filter effect, we verified it with a fluorescence spectrophotometer. First, we explored the optimal incubation time for detection of ALP. Add an ALP solution with an activity of 5 U / L to a solution containing 1 mM PNPP and 0.1 µM MgSO 4 Molybdenum disulfide quantum dots were incubated in alkaline buffer solution (Tris–HCl, 10 mM, pH = 8.0) at 37°C, and the fluorescence intensity of the solution under excitation at a wavelength of 400 nm was recorded by a fluorescence spectrophotometer every 3 min. Depend on Figure 5 It can be seen that the reaction reached a plateau after 60 min, so 60 min was selected as the optimal incubation time. Second, we explored the linear range of detection of ALP. ALP solutions (0 – 40 U / L) with different activities were added to the solution containing 1 mM PNPP and 0.1 µM MgSO 4 Molybdenu...

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Abstract

The invention belongs to the field of nano-particles, and particularly relates to a method for detecting alkaline phosphatase activity by molybdenum disulfide quantum dot inner-filter effect fluorescence. The method includes the steps: firstly, preparing fluorescent molybdenum disulfide quantum dots: taking molybdenum disulfide powder as a raw material, taking ethyl alcohol/water as a solvent, taking inorganic base as a stripping auxiliary and synthesizing the molybdenum disulfide quantum dots by an ultrasonic method; secondly, adding alkaline phosphatase solution into 4-nitrophenyl phosphatedisodium salt (PNNP) and magnesium sulfate alkaline buffer solution, and performing co-incubation; thirdly, adding the alkaline buffer solution into molybdenum disulfide quantum dot solution and performing further co-incubation; fourthly, building a standard work curve for detecting alkaline phosphatase; fifthly, acquiring the activity of the alkaline phosphatase in the solution to be detected according to a standard curve equation. The fluorescent molybdenum disulfide quantum dots have the advantages of stable performance, simple synthesis conditions, low price and the like, and are simple indetection process and high in selectivity and sensitivity when being used for detecting the alkaline phosphatase.

Description

technical field [0001] The invention belongs to the technical field of nanometer materials, and in particular relates to a method for fluorescently detecting alkaline phosphatase activity by utilizing the internal filter effect of molybdenum disulfide quantum dots. technical background [0002] Alkaline phosphatase (ALP), a common hydrolytic enzyme in mammalian tissues, has been widely used as an important biomarker for clinical diagnosis. ALP mostly exists in liver and gallbladder, kidney, bone and placenta, and is used to catalyze the dephosphorylation process of proteins, nucleic acids, sugars and alkaloids. The content of ALP in normal human serum is between 20-140U / L. The abnormal level of ALP in serum is closely related to breast cancer, prostate cancer, bone diseases, liver dysfunction and diabetes. Therefore, it is necessary to establish a sensitive method for quantitative detection of ALP. The determination of ALP activity today mainly includes isotope labeling, c...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/6486
Inventor 易涛钟亚平薛峰峰魏鹏李若涵曹春艳
Owner FUDAN UNIV
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