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Method for calibrating three-dimensional fluorescence

A technology of three-dimensional fluorescence and fluorescence intensity, which is applied in the field of three-dimensional fluorescence detection system and can solve the problems such as the decrease of fluorescence intensity

Inactive Publication Date: 2011-06-08
CHINESE RES ACAD OF ENVIRONMENTAL SCI
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Problems solved by technology

It can be seen that different fluorescent substances have different fluorescence characteristics and concentration ranges in which characteristic fluorescence occurs, but when the concentration is high, inner filtering and self-absorption (reabsorbance) phenomena will occur, resulting in a decrease in fluorescence intensity

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  • Method for calibrating three-dimensional fluorescence
  • Method for calibrating three-dimensional fluorescence
  • Method for calibrating three-dimensional fluorescence

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[0026] The chemical reagents used in this experiment are all superior grade or analytical grade. Humic acid was purchased from Tianjin Jinke Fine Chemical Research Institute. In order to keep its inherent fluorescence characteristics unaffected, the samples were not purified before use. Use 0.01mol / L NaOH solution to dissolve, use milli-Q ultrapure water (Millipore, 18.2MΩcm) to prepare 4 concentrations of humic acid (5mg / L, 10mg / L, 50mg / L, 100mg / L), use 0.01mol / LNaOH solution or 0.01mol / LHCl solution to adjust the pH value to 6.5, and then filter with GF / F glass fiber filter membrane (preliminarily burned at 450℃ for 5min) to measure the fluorescence intensity. In this study, a high-sensitivity Hitachi fluorescence spectrophotometer (HITACHI F-7000, Japan) with a 1cm quartz fluorescence sample cell was used for three-dimensional fluorescence excitation-emission matrix spectroscopy (3DEEM) measurement. The excitation light source is a 150W xenon arc lamp, the photomultiplier...

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Abstract

The invention discloses a method for calibrating three-dimensional fluorescence, which comprises: setting three-dimensional fluorescence measurement conditions, determining excitation-emission matrix spectra of a sample and obtaining a spectrogram and an excitation wavelength-emission wavelength- fluorescence intensity data matrix; testing the absorption of light of a corresponding wavelength of the sample by using a calibrating and testing instrument to obtain a transmittance (T) and an absorbency (A); and calibrating three-dimensional fluorescence data by dividing the fluorescence intensity(If) corresponding to each emission wavelength in a three-dimensional fluorescence matrix by the transmittance (T), tested by the calibrating and testing instrument, corresponding to the emission wavelength of the sample, or performing calibration by using the absorbency (A) according to a formula of If / 10-A. The calibration method is particularly suitable for testing high-concentration samples to be tested. In addition, the invention also discloses a testing system using the method.

Description

Technical field [0001] The invention relates to a method for three-dimensional fluorescence detection and correction, and a three-dimensional fluorescence detection system using the method. Background technique [0002] Three-dimensional fluorescence spectroscopy is a new analysis technology developed on the basis of fluorescence spectroscopy in the 1980s. The method of obtaining three-dimensional fluorescence data is generally to continuously scan the emission spectra at different excitation wavelength positions, and various drawings can be used. The software visualizes it in the form of isometric three-dimensional fluorescence projection (Ex-Em-If) or contour spectrum (Ex-Em). The application of fluorescence spectroscopy technology to study fluorescent substances is based on the structure containing a large number of aromatic ring structures with various functional groups and unsaturated aliphatic chains (the reason why substances have fluorescence is due to the low energy in t...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 刘俐李发生颜增光曹云者
Owner CHINESE RES ACAD OF ENVIRONMENTAL SCI
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