CN-116626005-B - Method for rapidly detecting CTC

Abstract

The invention discloses a method for rapidly detecting CTC, and belongs to the technical field of tetracycline detection. The fluorescence intensity of the solution to be detected, which is added with the fluorescent probe PEI-CuNCs and the surfactant, is measured by adopting a fluorescence spectrometer, and compared with the fluorescence intensity of PEI-CuNCs, if a new blue fluorescence emission band appears at 425nm and blue-green fluorescence at 500nm is reduced, the solution to be detected contains CTC components. The detection method has the advantages of simple and rapid operation, high sensitivity, good selectivity, low detection limit, low cost and the like, does not need large-scale instruments and professional operators, has good labeling recovery rate in the detection of an actual water sample, and has reliable detection result and good practicability.

Inventors

• LI TAIHUA
• Qian Meiru
• WANG ANQI
• DAI JING
• YANG JIN
• HAN JIANGANG
• JIN LONG

Assignees

• 南京林业大学

Dates

Publication Date

20260505

Application Date

20230526

Claims (3)

1. 1. A method for rapidly detecting CTC is characterized in that a fluorescence spectrometer is adopted to measure the fluorescence intensity of a solution to be detected, which is added with a fluorescence probe PEI-CuNCs and a surfactant, and compared with the fluorescence intensity of PEI-CuNCs, if blue-green fluorescence at 500nm is reduced and a new blue fluorescence emission band appears at 425nm, the solution to be detected contains CTC components; The method comprises the following steps: 1) The preparation method of the fluorescent probe PEI-CuNCs comprises adding Cu (NO 3 ) 2 solution into PEI solution, stirring at room temperature for 10min, adding ascorbic acid solution, performing temperature-controlled dark reaction, cooling to room temperature, and filtering with a microporous filter of 0.45 μm to obtain dark brown PEI-CuNCs solution; 2) Drawing a CTC standard concentration gradient curve; Preparing CTC solutions with different concentrations, respectively adding the CTC solutions into 20mmol/L acetic acid buffer solution containing CTAB with mass concentration of 0.1% and PEI-CuNCs with volume percentage of 1%, and measuring the fluorescence intensity of the system by using a fluorescence spectrometer after light-shielding reaction at room temperature, and drawing a linear relation graph by using a difference value delta F 425 /F 500 of the fluorescence intensity ratio F 425 /F 500 , wherein a linear regression equation of the CTC is y=0.4292x+0.1482; 3) Detecting a system to be detected; and (3) measuring the delta F 425 /F 500 value of the system to be measured by adopting the detection method same as that of the step (2), and calculating the concentration of the CTC in the system to be measured by using the curve of the step (2).
2. 2. The method for rapid detection of CTCs according to claim 1, wherein the linear concentration of CTCs ranges from 0 to 1 μmol/L and the detection limit is 0.96nmol/L.
3. 3. The method for rapid detection of CTCs according to claim 1, wherein in step 1), the temperature is controlled to 80 ℃ and the reaction is performed for 8 hours under a dark condition at 200 rpm.

Description

Method for rapidly detecting CTC Technical Field The invention belongs to the technical field of tetracycline detection, and particularly relates to a method for rapidly detecting aureomycin (CTC). Background Aureomycin (CTC), which was the earliest tetracycline antibiotic (TCs) discovered, is also known as "chlorotetracycline" and has inhibitory effects on most gram-positive bacteria, gram-negative bacteria, rickettsiae, and the like. CTC is mainly distinguished from TCs by Cl in the 7-position, which makes it more permeable to biological membranes, more widely available, more toxic and more easily absorbed by cells. CTCs have cross-resistance with other antibiotics of TCs, and due to the wide application of TCs, clinically common pathogenic bacteria also have stronger resistance to CTCs. At present, TCs in the environment mainly originate from livestock and poultry cultivation, hospitals, pharmaceutical factory pollution discharge and the like. CTCs do bring some benefits to human and animal health, however, the use of unconditioned CTCs poses a significant hazard, and CTCs eventually remain in the human body due to accumulation of food chains, posing a threat to human health. TCs can be deposited in bone and tooth tissues and combined with calcium deposited in newly formed bone and teeth to cause yellow deposition of teeth, and in addition, TCs can be enriched in liver, stomach and intestine tissues to cause tissue damage and can also cause anaphylactic reaction and teratogenesis, so that the establishment of a method for detecting CTC is very important. Various methods for quantitative analysis and detection of CTCs have been established, including Thin Layer Chromatography (TLC), high Performance Liquid Chromatography (HPLC), ultraviolet-visible spectroscopy (UV-Vis), mass Spectrometry (MS), or immunoassay. However, most of these methods are expensive, require high demands on operators, or are simple to operate, inexpensive, but have insufficient sensitivity. Therefore, it is highly necessary to develop a more sensitive CTC detection method with good selectivity and low detection cost. CTCs themselves have optical properties such as uv/visible light absorption, fluorescence, quantum yield, etc. The position and intensity of the fluorescence spectrum of the same CTC may be significantly different in different solvents, and if the solvent and the fluorescent substance form a complex or the solvent changes the ionization state of the fluorescent substance, the fluorescence spectrum may also change, and when a surfactant is added to the solution of the fluorescent substance, the fluorescence spectrum of the fluorescent substance may be enhanced, and the fluorescence spectrum of the fluorescent substance may be sensitized, based on the solubilization principle, deprotonation and other actions of the surfactant. Surfactants have found wide application in various fields, but they have not been studied much in terms of analytical detection of substances, particularly CTCs. Disclosure of Invention Aiming at the problems in the prior art, the invention aims to provide a method for detecting CTC, which is simple, quick, high in sensitivity and good in specificity. In order to solve the technical problems, the technical scheme adopted by the invention is as follows: a method for quickly detecting CTC includes such steps as measuring the fluorescent intensity of the solution to be detected, which contains PEI-CuNCs as fluorescent probe and surfactant, by fluorescent spectrometer, and comparing it with PEI-CuNCs, if the blue-green fluorescence at 500nm is decreased and a new blue fluorescent emission band appears at 425nm, the solution to be detected contains CTC component. According to the method for rapidly detecting CTC, the volume percentage of PEI-CuNCs added into a system to be detected is 1%. According to the method for rapidly detecting CTC, the surfactant added in the system to be detected is CTAB. According to the method for rapidly detecting CTC, the concentration of CTAB is 0.1% by mass. According to the method for rapidly detecting CTC, the pH value of a system to be detected is controlled to be 6. A method for rapidly detecting CTC comprises the following steps: 1) Preparing a fluorescent probe PEI-CuNCs; 2) Drawing a CTC standard concentration gradient curve; Preparing CTC solutions with different concentrations, respectively adding CTAB and PEI-CuNCs, performing light-shielding reaction at room temperature, and measuring the fluorescence intensity of the system by using a fluorescence spectrometer, and drawing a linear relation graph by using a difference value delta F 425/F500 of a fluorescence intensity ratio F 425/F500, wherein a linear regression equation of the CTC is y=0.4292x+0.1482; 3) Detecting a system to be detected; and (3) measuring the delta F 425/F500 value of the system to be measured by adopting the detection method same as that of the step (2), and calculating the concentration of t