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A method for measuring erythrine concentration based on fluorescence quantitative method

A fluorescence quantitative and concentration measurement technology, applied in fluorescence/phosphorescence, material excitation analysis, etc., can solve problems such as inability to solvent extraction and separation, and achieve the effect of rapid mass detection, short pretreatment time, and avoidance of influence.

Inactive Publication Date: 2018-03-23
GUANGXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When they exist alone, the fluorescence intensity of each has a good linear relationship with its concentration, but the fluorescence spectra of the three overlap almost completely. When the excitation wavelength is 280nm, the emission wavelength is 350nm, so when the three are mixed, they cannot be directly quantified by fluorescence spectroscopy. Although the auxin can be separated with an organic solvent (ethyl acetate) according to the hydrophilicity of the three, the hydrophilicity of erythroline and tryptophan is very close, and it cannot be extracted and separated with common solvents. , tryptophan became an impurity that was difficult to remove during the quantitative process of erythroline

Method used

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  • A method for measuring erythrine concentration based on fluorescence quantitative method
  • A method for measuring erythrine concentration based on fluorescence quantitative method
  • A method for measuring erythrine concentration based on fluorescence quantitative method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] 1. Draw a tryptophan standard curve

[0072] Prepare a 10 mg / L tryptophan standard solution; take 6 test tubes, one as a blank control, and add 1 μL, 2 μL, 3 μL, 4 μL, 5 μL of tryptophan standard solution dropwise respectively; add 100 μL of pH7 .4 disodium hydrogen phosphate / sodium dihydrogen phosphate buffer solution; be fixed to 1ml with distilled water, measure its fluorescence intensity respectively (excitation wavelength 280nm, emission wavelength 350nm), the result is as shown in Table 1, the blank control group (distilled water) Fluorescence intensity Z 0 =489; take the tryptophan mass concentration A as the abscissa, and the relative fluorescence intensity T' after subtracting the blank as the ordinate, draw a tryptophan standard curve, and the results are as follows image 3 shown.

[0073] A (μg / L)

10

20

30

40

50

T

526

587

623

675

756

[0074] Table 1

[0075] 2 Preparation of standard mixed solution

[0...

Embodiment 2

[0112] Determination of Erythrine and Tryptophan in the Extract of Colored Bean Puffball Mycelia

[0113] The colored bean puffball is a fungus that can synthesize erythrine. Its extract is first extracted with chloroform and ethyl acetate to remove indole acetic acid. Take 900 μL of the aqueous phase, add 100 μL of pH 7.4 disodium hydrogen phosphate-sodium dihydrogen phosphate buffer solution dropwise, and perform a test (excitation wavelength 280 nm, emission wavelength 350 nm), and the measured fluorescence intensity is X=3081, which is located in the present invention. scope of application. After adding 1 mL of formaldehyde dropwise, let stand for 1 hour, and measure the fluorescence intensity Y=2013. blank controlX 0 = 419, Y 0 =525; Substituting into the formula can get, the concentration of erythrine is 38.71 μg / L, and the mass in 900 μL test solution is 3.87×10 -8 g.

[0114] The measurements in the above examples all use a Hitachi F-7000 fluorescence spectrophoto...

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Abstract

The invention discloses a method for measuring the concentration of erythrotophanine based on a fluorescence quantitative method under the condition that tryptophan impurities exist, comprising the following steps: preparing a tryptophan standard solution and drawing a tryptophan standard curve; preparing a tryptophan Mix the standard solution with Erythrinaline, measure the fluorescence intensity of the standard mixed solution, draw the standard curve of the standard mixed solution; calculate the mass concentration of tryptophan in the mixed standard solution, draw the linear relationship curve 1, and draw the linear relationship curve 2 , deduce the calculation formula of the concentration of erythrine, and determine the concentration of erythrine solution. This method can directly quantify the fluorescence of erythrine under the condition of tryptophan impurity with fluorescence spectroscopy, avoiding color The impact of amino acid impurities on the test results, this method has short pretreatment time, low detection cost, and can be quickly detected in large quantities.

Description

technical field [0001] The invention belongs to the field of molecular measurement, in particular to a method for measuring the concentration of erythroline based on a fluorescence quantitative method. Background technique [0002] Xiazhen Erythrine, also known as Hepa Erythrine, CAS No. 487-58-1, Molecular Formula C 14 h 18 N 2 o 2 , is a common alkaloid in plants of the genus Acacia and Erythrina in the family Fabaceae. Erythrine can significantly inhibit mouse ear swelling, reduce mouse ear weight; increase mouse thymus, spleen weight, improve mouse serum hemolysin level; significantly reduce CCl 4 and naphthyl isothiocyanate-induced liver injury mice serum transaminase activity and bilirubin content, to protect the liver injury. Experiments on mice suggest that erythracine has obvious anti-inflammatory, immune-enhancing, enzyme-reducing, jaundice-reducing, and liver-injury effects. One of the hormones, auxin, is similar and contains indole groups, but it is an auxi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 陆祖军刘祎李云飞梁士楚
Owner GUANGXI NORMAL UNIV
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