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A method for rapid detection of lead ions
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A lead ion, fast technology, applied in the direction of material analysis by observing the influence of chemical indicators, and analysis by making materials undergo chemical reactions, etc., can solve the problems of complex equipment, time-consuming, expensive, etc., and achieve low cost, The effect of high sensitivity and easy operation
Active Publication Date: 2019-04-02
SHENYANG PHARMA UNIVERSITY
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Embodiment 1
[0036] 1. Preparation of gold nanoparticles:
[0037] Add 100mL of distilled water to a 250mL three-necked flask, then add 750μL of sodium citrate aqueous solution (1g / 10mL), stir and heat to 100℃, then add 2mL HAuCl 4 (0.09424M), keep at 100°C, continue the reaction for 30 minutes, and cool to room temperature. The solution turns red and the preparation is complete. Deionized water washes away excess impurities, centrifuges, and then disperses in deionized water.
[0038] 2. Preparation of lead specific binding protein:
[0039] (1) Preparation of α2-microglobulin (a2m):
[0040] a. Construction of pET28a-a2m expression vector
[0041] According to the preference of E. coli codons and the amino acid sequence of the target protein, the target gene was fully synthesized by Suzhou Jinweizhi Biotechnology Co., Ltd. The sequence is as follows:
[0069] Take 9mL CTAB stock solution 0.1M, 250μL chloroauric acid stock solution, 600μL sodium borohydride solution and mix, and react at 25°C for 2 hours to prepare seed solution.
[0070] Take 40mL CTAB stock solution, add 2mL chloroauric acid stock solution, 400μL silver nitrate solution (0.01M), 300μL ascorbic acid solution (0.1M), 800μL hydrochloric acid, and 10μL seed solution. The reaction was carried out at 37°C for 12 hours to prepare rod-shaped gold nanoparticles.
[0071] Take the rod-shaped gold nanoparticle solution prepared above, pipet 2 mL and place it in an EP tube. Centrifuge at 8000rpm / 10min, and remove the supernatant. Add buffer solution to disperse separately, then add 1 mL of the lead-binding protein solution prepared above, and place it in an ultrasonic cleaner to sonicate for 10 min. Place it at room temperature to make the gold nanorods fully bond with the lead binding protein. Centrifuge to remove unbound protein.
[0072] Add different amounts of lead i...
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Abstract
The invention belongs to the technical field of heavy metal detection, and relates to a method for on-line and quick detection of heavy metal lead ions. In the method, gold nanoparticles are used as a signal sensing material, protein which has a specific recognition function on the lead ions is used as a lead ion ligand, and the purpose of detection of the lead ions is realized through change of resonance strength and wavelength of plasmas on the surfaces of the gold nanoparticles, which is caused by the coordination process of the ligand and the lead ions. The method is mild in reaction condition, simple and safe in operation, high in repeatability, capable of eliminating inconvenience caused by the fact that the conventional lead ion detection pretreatment process is complex, and lots of instrument and equipment operations are involved, excellent in specificity, and capable of being widely used for quick and convenient detection of the metal lead ions in food, cosmetics and environments.
Description
Technical field [0001] The invention belongs to the technical field of heavy metal detection, and relates to a method for online and rapid detection of heavy metal lead ions. Based on gold nanoparticles, an optical probe with specific recognition function for heavy metal lead ions was constructed. Through the specific binding of lead binding protein and lead ions, a colorimetric analysis method based heavy metal lead ions was successfully constructed. The sensor realizes rapid and specific detection of lead ions. Background technique [0002] With the advancement of science and technology and the development of industry, environmental pollution and food safety problems have emerged and become increasingly aggravated. Heavy metal pollution seriously threatens people's health and ecological environment, so heavy metal pollution has become one of the most important issues recently. Design-sensitive tools and invented detection methods have attracted great attention, and many metho...
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