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Fluorescence detection method for amino acid under different pH values

A fluorescent detection and amino acid technology, applied in the fields of analytical chemistry and life sciences, can solve the problems of amino acid molecular detection that has not been reported yet, few high-selective identification detection, and difficulty in rapid detection, etc., and achieves wide application prospects and use value, operation The method is simple and the effect with superior performance is selected

Inactive Publication Date: 2013-12-25
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have the advantages of high precision and low detection limit, but the long sample pretreatment time and high cost make it difficult to adapt to rapid detection, which limits their application
In recent years, due to the high sensitivity and selectivity of fluorescence analysis, real-time in situ detection, and simple equipment, a variety of fluorescent molecular sensors for amino acid identification have been reported one after another. However, most fluorescent molecular sensors can only selectively recognize some thiol-containing Amino acid molecules with special functional groups, but there are relatively few highly selective recognition and detection of other amino acid molecules
Especially the detection of amino acid molecules under different pH conditions has not been reported yet.

Method used

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  • Fluorescence detection method for amino acid under different pH values
  • Fluorescence detection method for amino acid under different pH values
  • Fluorescence detection method for amino acid under different pH values

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1: Fluorescent Probes P Synthetic method:

[0017] Weigh 28 mg (0.02 mmol) of seven-membered melon ring and 4.4 mg (0.02 mmol) of acridine hydrochloride respectively, add them to 50ml of distilled water, and stir and react at 80-95°C for 2h. After cooling to room temperature, the aqueous solvent was removed by rotary evaporation. Precipitate with acetone, filter with suction, and dry. 30 mg of white solid was prepared with a yield of 94%.

[0018] Preparation of buffer solution: Acetic acid-sodium acetate buffer solution (pH4.6) Take 5.4g of sodium acetate, add 50ml of water to dissolve it, adjust the pH value to 4.6 with glacial acetic acid, and then dilute to 100ml with water. Phosphate buffer (pH7.2) Take 50ml of 0.2mol / L potassium dihydrogen phosphate solution and 35ml of 0.2mol / L sodium hydroxide solution, add freshly boiled cold water to dilute to 200ml, shake well, and get ready. pH 2.0 is directly diluted with 0.1 mol / L dilute hydrochloric acid and d...

Embodiment 2

[0019] Example 2: Fluorescent Probes P Identification and detection of amino acid molecules

[0020] (1) Fluorescent probes P Solution preparation method: Weigh 16.2 mg fluorescent probe solids, dissolve them in different pH buffers, and prepare 10 mL solutions with a concentration of 1.00×10 -3 mol L -1 , according to the need to use different pH buffers to dilute to the appropriate concentration step by step.

[0021] (2) Standard solution of amino acid molecules: weigh high-grade pure amino acid molecules to prepare 100mL solution, the concentration of amino acid molecules is 1.00×10 -3 mol L -1 , as needed, use different pH buffer solutions to gradually dilute to an appropriate concentration;

[0022]Add fluorescent probe to 10.0 mL volumetric flask P Different pH buffers (1.00×10 -4 mol L -1 , 1.0 mL), tryptophan or lysine molecule (2.00×10 -4 mol L -1 , 1.0 mL). Dilute to the mark with different pH buffer solutions, shake well, let stand at room temperature...

Embodiment 3

[0027] Weigh 14 mg of seven-membered melon ring and 2.5 mg of acridine hydrochloride, dissolve them in a buffer solution with pH=7.2, and prepare 10.0 mL with a concentration of 1.0 × 10 -3 mol L -1 , serially diluted to 1.00 × 10 with buffer as needed -4 mol L -1 (2) Standard solution of tryptophan or lysine and other coexisting amino acid molecules: weigh superior pure amino acid molecules to prepare 100mL buffer solution with pH=7.2, the concentration of amino acid molecules is 1.00×10 -3 mol L -1 , and gradually dilute to a suitable concentration with buffer solution as needed; (3) Take five 10.0 mL volumetric flasks and add fluorescent probe 1.00×10 -4 mol L -1 Standard solution 1.0 mL, add 1.00×10 -4 mol L -1 , 0, 0.2, 0.5, 1, 2 ml of tryptophan or lysine solution diluted to the mark, shake well, and stand at room temperature for 5 minutes; (4) introduce fluorescence spectrum for measurement, excitation wavelength 245nm; (5) tryptophan The concentration of ac...

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Abstract

The invention relates to a fluorescence detection method for amino acid under different pH values. Under an aqueous solution medium condition, by utilizing a host-guest fluorescent probe and employing fluorescence emission spectrometry and a visual method, lysine or tryptophan or a mixture of lysine and tryptophan can be detected and identified under different pH values with high sensitivity and high selectivity.

Description

technical field [0001] The invention relates to a fluorescent detection method for amino acids at different pH values, belonging to the fields of analytical chemistry and life sciences. Background technique [0002] Amino acids are the basic substances that make up living organisms and the basic substances of cell metabolism. The rapid identification of amino acids is of great significance to the analysis of cell metabolism, so the selective identification and detection of amino acids has aroused great interest. At present, the commonly used methods for identifying amino acids are high performance liquid chromatography (HPLC), gas chromatography (GC), capillary, electrophoresis (CE) and other chromatographic methods. These methods have the advantages of high precision and low detection limit, but the sample pretreatment time is long, the cost is high, and it is difficult to adapt to rapid detection, which limits their application. In recent years, due to the high sensitivi...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 宋森倪新龙陶朱
Owner GUIZHOU UNIV
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