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Method for quantitatively detecting alkaline phosphatase in seawater based on surface enhanced Raman spectroscopy technology

A surface-enhanced Raman and spectroscopic technique, applied in the field of Raman spectroscopic detection, can solve the problems of complex experimental steps and complex sample pretreatment.

Pending Publication Date: 2021-08-24
SHANGHAI OCEAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the detection methods of ALP activity include fluorescence method, electrochemical analysis method, etc. Although these methods can complete the detection of ALP activity, they are limited by the shortcomings of complex sample pretreatment and complicated experimental steps. Fast and efficient ALP detection method

Method used

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  • Method for quantitatively detecting alkaline phosphatase in seawater based on surface enhanced Raman spectroscopy technology
  • Method for quantitatively detecting alkaline phosphatase in seawater based on surface enhanced Raman spectroscopy technology
  • Method for quantitatively detecting alkaline phosphatase in seawater based on surface enhanced Raman spectroscopy technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Quantitative Model

[0026] 11 900μL of different activities (10U / mL, 5U / mL, 1U / mL, 0.5U / mL, 0.1U / mL, 50mU / mL, 10mU / mL, 5mU / mL, 1mU / mL, 0.5mU / mL, 0.1mU / mL) ALP solution was mixed with 1mg / mL and 100μL BCIP solution and incubated for 2h, respectively, after adding 20% ​​DMSO solution, the respective SERS signals were measured, and the obtained spectrum was as follows: image 3 shown.

[0027] from image 3 It can be seen that there is no direct linear relationship between the SERS characteristic peak intensity of the product BCI oxidized dimer and the ALP concentration, because the intensity of the Raman spectrum is affected by factors such as laser power stability, enhanced reagent uniformity, and solvent background noise. It is difficult to directly use the intensity of the characteristic peaks of the Raman spectrum to perform quantitative analysis. Therefore, the added DMSO solvent was added at 677 cm -1 The characteristic peaks of the Raman spectrum nea...

Embodiment 2

[0032] Example 2 Seawater verification test

[0033] Fresh seawater samples were collected from the East China Sea (30°39'48"N, 122°29'48"E) in December 2020. The sample is ocean surface seawater, which is directly sampled by fishing boats. Mix 900 μL fresh seawater sample with 1 mg / mL and 100 μL BCIP solution for 2 h, add 20% volume DMSO solution and measure the SERS signal, the obtained spectrum is as follows Figure 5 shown.

[0034] like Figure 5 as shown, 600cm in the picture -1 There are obvious Raman spectral peaks at -1The Raman spectral intensity at the wavenumber reaches 6389.6, which is due to the Raman spectral peaks caused by the C-S-C symmetrical stretching vibration in DMSO. The ratio of the two peaks is 0.377. This value is substituted into the above model to realize the quantitative detection of the ALP activity of the seawater sample. The ALP activity of the water sample obtained was equivalent to 0.5 mU / mL E. coli ALP activity.

[0035] Using BCIP as ...

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Abstract

The invention provides a method for quantitatively detecting alkaline phosphatase based on a surface enhanced Raman spectroscopy technology by taking BCIP as a substrate and DMSO as an internal standard substance. Results show that the ALP activity and the intensity ratio (600cm <-1 > / 677cm <-1 >) of the characteristic peak to the internal standard peak have a good linear relationship, and the correlation coefficient is 0.977; the ALP activity in a seawater sample is successfully and quantitatively detected by using the model, and rapid detection of the ALP activity in seawater is realized. Meanwhile, the method can also be applied to activity detection of extracellular enzymes of other microorganisms in seawater, and a solid scientific foundation is laid for in-situ detection of the activity of the extracellular enzymes of the microorganisms in the seawater.

Description

technical field [0001] The invention relates to a method for quantitatively detecting alkaline phosphatase in seawater based on surface-enhanced Raman spectroscopy technology, and belongs to the technical field of Raman spectroscopy detection. Background technique [0002] Alkaline Phosphatase (ALP) is widely distributed in the marine environment and can participate in the hydrolysis reaction of phosphate compounds. It is transported into microbial cells and provides energy sources for the life activities of microorganisms. Many literature data show that bacteria and algae rely on the nutrients produced by the hydrolysis of ALP to survive. The synthesis of marine microbial ALP is affected by the surrounding water environment. When the surrounding water environment is deficient in inorganic phosphorus, the microorganisms will use the inorganic phosphorus in their own cells to maintain life activities; when the inorganic phosphorus concentration in the surrounding water enviro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/65
CPCG01N21/658C12Q1/42C12Q2334/52G01N2001/2893G01N1/28G01N33/18C12Y301/03001
Inventor 吴勇张宏鸽方家松魏玉利曹军伟李思聪
Owner SHANGHAI OCEAN UNIV
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