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Melamine fluorescence spectrum detection method based on metallic ruthenium polypyridine complex

A metal ruthenium polypyridine and melamine technology, applied in the field of melamine detection, can solve the problems of less melamine, kidney stones, malnutrition, etc., and achieve the effects of easy control, simple steps, and fast process.

Inactive Publication Date: 2015-07-22
TONGJI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Melamine cannot be metabolized in the body. Excessive intake of this food will not only cause kidney stones, but also cause malnutrition due to insufficient protein intake.
At present, there are relatively few reports on the detection of melamine by fluorescence spectroscopy

Method used

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  • Melamine fluorescence spectrum detection method based on metallic ruthenium polypyridine complex
  • Melamine fluorescence spectrum detection method based on metallic ruthenium polypyridine complex
  • Melamine fluorescence spectrum detection method based on metallic ruthenium polypyridine complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Fluorescent detection of melamine standard solution

[0043] (1) Interaction of rich T DNA and melamine

[0044] Dissolve the melamine to be tested in methanol to prepare standard test solutions of melamine with different gradients. Add 15 μL T30 (concentration: 100 μmol / L, sequence from 5' to 3': TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT) into 15 centrifuge tubes (1.5mL) with a pipette gun, and then add different concentrations of melamine in sequence with a pipette gun. The test solution was diluted to 985 μL with 10 mmol / L Tris-HCl (pH 7.0) buffer. The mixture was incubated at room temperature for 15 minutes. T-rich DNA can form special three hydrogen bonds with melamine, thus forming a three-stranded structure. The T-rich DNA changes from a single-stranded structure to a triple-stranded structure.

[0045] (2)[Ru(phen) 2 dppz-idzo] 2+ Insert triplex structure

[0046] 15μL [Ru(phen) 2 dppz-idzo] 2+ (200 μmol / L) was added to the above mixture, and the system was incu...

Embodiment 2

[0052] selective research

[0053] (1) Cultivation of T-rich DNA sequences and interfering substances

[0054] Use a pipette gun to add 15 μL T30 (concentration: 100 μmol / L, sequence from 5' to 3': TTTTTTTTTTTTTTTTTTTTTTTTTTTTTT) into each centrifuge tube (1.5 mL), and then use a pipette gun to add a certain concentration of interfering substances ( Glycine, Histidine, Threonine, Valine, Lysine, Cysteine, Glucose, Ca 2+ , Mg 2+ , Zn 2+ , dicyandiamine, uracil, ATP, BSA) and melamine, diluted to 985 μL with 10 mmol / L Tris-HCl (pH 7.0) buffer. The mixture was incubated at room temperature for 15 minutes.

[0055] (2)[Ru(phen) 2 dppz-idzo] 2+ insert

[0056] 15μL [Ru(phen) 2 dppz-idzo] 2+ (200 μmol / L) was added to the above mixture, and the system was incubated at room temperature for 15 minutes. The final concentration of T30 is 1.5μmol / L, [Ru(phen) 2 dppz-idzo] 2+ The final concentration is 3.0 μmol / L. The final concentration of interfering substances was 10 μmol / L...

Embodiment 3

[0061] Sample spike recovery experiment

[0062] (1) Milk sample pretreatment

[0063] Add 2mL of pure milk and 1mL of melamine solutions of different concentrations into a 10mL centrifuge tube, add 2mL of triple distilled water, then add 1mL of acetonitrile and 1mL of methanol, vortex and shake for 1min to remove protein, and the mixture is ultrasonicated for 10min. Centrifuge for 10min. Filter with a 0.22 μm microporous membrane, take the supernatant, transfer to another centrifuge tube, and set aside.

[0064] (2) Culture of T-rich DNA sequences and milk samples

[0065] Use a pipette gun to add 15 μL T30 (concentration: 100 μmol / L, sequence from 5' to 3': TTTTTTTTTTTTTTTTTTTTTTTTTTTTTT) into each centrifuge tube (1.5 mL), and then use a pipette gun to sequentially add milk containing a certain concentration of melamine The sample was diluted to 985 μL with 10 mmol / L Tris-HCl (pH 7.0) buffer. The mixture was incubated at room temperature for 15 minutes.

[0066] (3)[Ru...

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Abstract

The invention relates to a melamine fluorescence spectrum detection method based on metallic ruthenium polypyridine complex. Melamine and DNA rich in thymine can form the effect of three special hydrogen bonds, so that the DNA can be transformed into three strands from a single strand, and as (Ru(phen)2dppz-idzo)<2+> (phen is 1,10-phenanthroline, dppz is bispyridine phenazine, and idzo is imidazolone) has strong insertion effect on three-strand DNA, fluorescence enhanced range of the three-strand DNA is obviously higher than that of single-strand DNA. When the concentration of the melamine is 0 to 7.0 micro mol / L, the concentration of the obviously and a fluorescence response value have a good linear relation, linear regression equations are respectively F=199.96+85.41c (C: micro mol / L), and a linear coefficient R2 is 0.9923. Meanwhile, by adding melamine with different concentrations to pure milk samples through a constructed method, a satisfactory recovery rate is obtained.

Description

technical field [0001] The invention relates to a method for detecting melamine, in particular to a method for detecting fluorescence spectrum of melamine based on metal ruthenium polypyridine complex. Background technique [0002] Melamine (C 3 h 3 N 6 ), is a white monoclinic crystal, commonly known as melamine or protein essence, is an important nitrogen-heterocyclic organic chemical raw material, and is usually widely used in fireproof materials. Melamine is not a food additive and is not allowed to be added to food and animal feed. However, in recent years, criminals have added melamine to infant milk powder in order to increase the nitrogen content of milk powder. Milk powder mixed with melamine has caused many infants to suffer from urinary system diseases, resulting in health problems for many infants and deaths. Usually, the detection method of protein content in food is the Kjeldahl method, which is to determine the protein equivalent by detecting the nitrogen ...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N1/28G01N1/38
Inventor 刘冰张艺伟王秀枝姚天明
Owner TONGJI UNIV
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