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Method for measuring content of dissoluble lignin by utilizing Coomassie brilliant blue G-250

A technology of Coomassie Brilliant Blue and G-250, which is applied in the measurement of color/spectral characteristics, etc., can solve the problems of interference in the determination of degradation products of carbohydrates, and achieve the effect of simple and fast measurement method, simple preparation, and avoidance of interference

Inactive Publication Date: 2015-08-05
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the disclosed spectroscopic determination method, it is mostly used to measure in the ultraviolet light region, which is likely to cause the interference of the degradation products of carbohydrates to the determination. After searching, the current method of using the wavelength of 595nm in the visible light region to measure the content of soluble lignin is not yet available. see the report

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  • Method for measuring content of dissoluble lignin by utilizing Coomassie brilliant blue G-250

Examples

Experimental program
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Embodiment 1

[0022] (1) Take the lignin solution to be tested, adjust the pH of the solution to 12 with 1M NaOH, and set aside;

[0023] (2) Centrifuge the spare solution in step (1) at 9000g for 5 minutes, then take the supernatant, and carry out 10×, 20×, 30×, 40×, 50× concentration gradient dilution, and take the diluted solution for each 100μl, add 5ml of Coomassie Brilliant Blue G-250 reagent, use a vortex mixer to mix for 15 seconds, to be tested;

[0024] (3) Take 100 μl of 0.1M NaOH, add 5ml of Coomassie Brilliant Blue G-250 reagent, and use a vortex mixer to mix for 15 seconds, as a test blank control;

[0025] (4) Take each 3ml of the sample to be tested in step (1) or the blank control in step (2), put it into a glass cuvette with a light path of 1cm, and measure the absorbance value at 595nm within 2 to 5 minutes;

[0026] (5) The absorbance value of the sample to be tested minus the absorbance value of the blank control is the net absorbance value, according to the standard c...

Embodiment 2

[0029] (1) Take the powdered solid lignin to be tested, first dissolve it with 0.1M NaOH to prepare its saturated solution, and adjust its pH to 13 for subsequent use;

[0030] (2) Centrifuge the spare solution in step (1) with 1000g for 5 minutes, then take the supernatant, carry out 10×, 20×, 30×, 40×, 50× concentration gradient dilution, take the diluted solution respectively 100μl, add 5ml of Coomassie Brilliant Blue G-250 reagent, use a vortex mixer to mix for 20 seconds, to be tested;

[0031] (3) Take 100 μl of 0.1M NaOH, add 5ml of Coomassie Brilliant Blue G-250 reagent, and use a vortex mixer to mix for 20 seconds as a test blank control;

[0032] (4) Take each 3ml of the sample to be tested in step (1) or the blank control in step (2), put it into a glass cuvette with a light path of 1cm, and measure the absorbance value at 595nm within 2 to 5 minutes;

[0033] (5) The absorbance value of the sample to be tested minus the absorbance value of the blank control is the...

Embodiment 3

[0036] (1) Take the lignin solution to be tested, adjust the pH of the solution to 11 with 1M NaOH, and set aside;

[0037] (2) Centrifuge the spare solution in step (1) with 8000g for 10 minutes, then take the supernatant, and carry out 10×, 20×, 30×, 40×, 50× concentration gradient dilution, and take the diluted solution respectively 100μl, add 5ml of Coomassie Brilliant Blue G-250 reagent, use a vortex mixer to mix for 17 seconds, to be tested;

[0038] (3) Take 100 μl of 0.1M NaOH, add 5ml of Coomassie Brilliant Blue G-250 reagent, and use a vortex mixer to mix for 17 seconds, as a test blank control;

[0039] (4) Take each 3ml of the sample to be tested in step (1) or the blank control in step (2), put it into a glass cuvette with a light path of 1cm, and measure the absorbance value at 595nm within 2 to 5 minutes;

[0040] (5) The absorbance value of the sample to be tested minus the absorbance value of the blank control is the net absorbance value, according to the sta...

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Abstract

The invention discloses a method for measuring content of dissoluble lignin by utilizing Coomassie brilliant blue G-250. The method comprises the following steps: taking to-be-measured lignin solution, regulating the pH value of the solution to be 11-13 by using 1M of NaOH solution, and centrifuging the stock solution by using 8000-1000g of NaOH solution; taking the supernatant, performing concentration gradient dilution, respectively adding a Coomassie brilliant blue G-250 reagent, measuring an absorbance value at the 595nm, finding out the lignin concentration value which corresponds to the net absorbance value according to the pre-drawn standard curve, thereby obtaining the lignin concentration value of the to-be-measured sample. The method disclosed by the invention is simple and rapid, the interference of degradation products of carbohydrates on the measurement result can be avoided during ultraviolet wavelength area measurement, and the method is applied to researching measurement of the lignin content and measurement of the lignin content in other lignin-containing samples in the lignin degradation experiment.

Description

technical field [0001] The invention relates to the determination of lignin content, in particular to a method for determining soluble lignin content by using Coomassie Brilliant Blue G-250. Background technique [0002] Most of the existing methods for determining lignin content are to measure lignin in solid lignocellulosic raw materials or pulp. Usually, the lignocellulosic raw materials or pulp are acidified with concentrated sulfuric acid to measure acid-soluble lignin and acid-insoluble lignin respectively. content. For acid-soluble lignin, measure the absorbance value at 205nm or 280nm, and calculate the content of acid-soluble lignin according to the empirical formula. However, carbohydrate hydrolyzates such as furfural produced in the sulfuric acid hydrolysis of lignocellulose also absorb in the ultraviolet region, which will interfere with the accuracy of lignin content determination. [0003] Existing methods for the determination of dissolved lignin include lig...

Claims

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Application Information

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IPC IPC(8): G01N21/31
Inventor 黄峰王芳芳艾明强张玉忠卢雪梅
Owner SHANDONG UNIV
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