CN111595989A - Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device - Google Patents

Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device Download PDF

Info

Publication number
CN111595989A
CN111595989A CN202010564997.9A CN202010564997A CN111595989A CN 111595989 A CN111595989 A CN 111595989A CN 202010564997 A CN202010564997 A CN 202010564997A CN 111595989 A CN111595989 A CN 111595989A
Authority
CN
China
Prior art keywords
solid
extraction
antibacterial agent
phase
graphene
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN202010564997.9A
Other languages
Chinese (zh)
Inventor
汪龙眠
沈曹凤
彭福全
庞晴晴
朱翔
张洪玲
罗一单
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Nanjing Institute of Environmental Sciences MEE
Original Assignee
Nanjing Institute of Environmental Sciences MEE
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Nanjing Institute of Environmental Sciences MEE filed Critical Nanjing Institute of Environmental Sciences MEE
Priority to CN202010564997.9A priority Critical patent/CN111595989A/en
Publication of CN111595989A publication Critical patent/CN111595989A/en
Pending legal-status Critical Current

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N30/08—Preparation using an enricher
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N30/14—Preparation by elimination of some components
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/74—Optical detectors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/86—Signal analysis
    • G01N30/8624—Detection of slopes or peaks; baseline correction
    • G01N30/8631—Peaks
    • G01N30/8634—Peak quality criteria
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/062—Preparation extracting sample from raw material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N2030/065—Preparation using different phases to separate parts of sample
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N30/14—Preparation by elimination of some components
    • G01N2030/143—Preparation by elimination of some components selective absorption
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N30/14—Preparation by elimination of some components
    • G01N2030/146—Preparation by elimination of some components using membranes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
    • G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
    • G01N2030/884—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample organic compounds

Abstract

The invention discloses a detection method of an antibacterial agent in a water body based on graphene solid-phase extraction and a solid-phase micro-extraction device, and belongs to the technical field of pollutant trace monitoring. The method comprises the following steps: 1) carrying out activation pretreatment on an adsorbent in a solid-phase micro-extraction device; 2) preparing an antibacterial agent standard solution, measuring by adopting HPLC and drawing a standard curve; 3) pretreatment of a water sample to be detected and detection of the content of an antibacterial agent: filtering and adjusting the pH value of a water sample to be detected to 7.0-7.5; extracting the antibacterial agent in the water sample to be detected by using a solid phase microextraction device; and desorbing the adsorbed solid-phase micro-extraction device and detecting to obtain the content of the antibacterial agent in the water sample. According to the method, the antibacterial agent in the water sample to be detected is extracted at high efficiency through the solid-phase micro-extraction device, and then the desorption is carried out for determination, so that the antibacterial agent can be effectively enriched and desorbed into the organic solvent, and the technical problem that the detection is inaccurate by adopting high performance liquid chromatography when the antibacterial agent in the water sample is not uniformly distributed or the content of the antibacterial agent is low is solved.

Description

Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device
Technical Field
The invention belongs to the technical field of pollutant trace monitoring, and particularly relates to a detection method of an antibacterial agent in a water body based on graphene solid-phase extraction and a solid-phase micro-extraction device.
Background
Triclosan (TCS) and Triclocarban (TCC) are antibacterial agents commonly used in personal care and household products. The annual world production of TCS and TCC exceeds 1500 and 450 tons respectively. They are widely added into washing and chemical products such as soap, toothpaste, disinfectants, mouth wash, shampoo and the like. Methyl triclosan (M-TCS) is one of the TCS degradation products produced by aerobic biodegradation. M-TCS is more lipophilic and therefore more persistent in the environment than TCS. These antibacterial agents may pose a hazard to the environment and human health due to the resistance and bioaccumulation of antibiotics.
Due to large-scale production and widespread use, TCS, TCC and M-TCS are widely distributed in the environment, especially in wastewater, rivers and sediments. The main source for introducing the compounds into the environment is sewage treatment plant drainage, in the sewage treatment plant, the highest average removal rates of TCS and TCC wastewater treatment are respectively 58-99% and 20-75%, and M-TCS is formed in the TCS microbial degradation process. Therefore, there is a need to develop suitable analytical methods to determine these compounds in environmental substrates, especially wastewater.
Generally, the wastewater is treated to have low contents of TCS, TCC and M-TCS, so that the pretreatment of the sample is important for purifying the sample from the matrix component and enriching the sample. The traditional sample pretreatment method comprises liquid-liquid extraction, Soxhlet extraction, chromatography, distillation, adsorption, centrifugation, filtration and the like, the established detection method mainly adopts chromatography, mainly adopts Gas Chromatography (GC) and High Performance Liquid Chromatography (HPLC), and the corresponding mass spectrum coupling technology (GC/MS or LC/MS) is also reported. The enrichment and purification treatment technology of the sample is mainly based on Solid Phase Extraction (SPE) procedures of different materials, so that the enrichment and purification treatment technology is suitable for analysis of different sample matrixes. As a sample processing technology which is very suitable for analyzing trace substances of an aqueous sample, the SPE has the biggest characteristics of low consumption of organic solvent and pretreatment time saving, and the technical core is an adsorbing material which determines the sensitivity and selectivity of SPE extraction.
However, the existing SPE consumes long time, needs a relatively expensive SPE chromatographic column, and has a plurality of problems that analytes may be lost due to multi-step operation in the extraction and evaporation processes, the extraction effect is not ideal, and the accuracy of the detection result is further influenced.
Disclosure of Invention
1. Problems to be solved
Aiming at the problem of inaccurate detection of the antibacterial agent content in the existing water body, the invention provides a detection method of the antibacterial agent in the water body based on graphene solid-phase extraction.
2. Technical scheme
In order to solve the problems, the technical scheme adopted by the invention is as follows:
a detection method of an antibacterial agent in a water body based on graphene solid-phase extraction comprises the following steps:
1) pretreatment of a solid phase micro-extraction device: activating an adsorbent in a solid phase micro-extraction device by using acetonitrile;
2) standard solutions were prepared and standard curves were plotted: dissolving an antibacterial agent in acetonitrile to obtain a corresponding stock solution, and adding water to dilute the stock solution into a series of standard solutions with concentration gradients;
directly injecting the standard solution into a high performance liquid chromatograph for analysis and detection to obtain a linear relation, namely a standard curve, of the concentration of the target compound to the response value of the chromatographic peak area;
3) pretreatment of a water sample to be detected and detection of the content of an antibacterial agent: filtering the collected water sample by using a filter membrane, and then adjusting the pH value of the water sample to be detected to 7.0-7.5 by using a phosphate buffer solution;
extracting the antibacterial agent in the water sample to be detected by using a solid phase microextraction device;
and (3) carrying out desorption treatment on the adsorbed solid-phase micro-extraction device, collecting desorption liquid, directly injecting the desorption liquid into a high performance liquid chromatograph for analysis and detection, and calculating the obtained measured value by contrasting with a standard curve to obtain the content of the antibacterial agent in the water sample to be detected.
Preferably, the solid-phase micro-extraction device is a micro solid-phase extraction device and comprises an inner-layer extraction membrane bag and an outer-layer extraction membrane bag, wherein the inner-layer extraction membrane bag is filled with graphene as an adsorbent;
the outer extraction membrane bag is sleeved outside the inner extraction membrane bag, and a sinker is arranged in the outer extraction membrane bag to realize the underwater positioning of the solid-phase micro-extraction device.
Preferably, the inner extraction film bag and the outer extraction film bag are prepared by three-side hot-sealing polypropylene films.
Preferably, the relationship between the amount of the graphene and the antibacterial agent content in the water sample to be extracted is as follows: when the concentration of the antibacterial agent is less than or equal to 1000 mug/L, 20mg of graphene is filled in the solid-phase micro-extraction device.
Preferably, the activation treatment in step 1) is specifically: and (3) soaking the solid phase micro-extraction device in acetonitrile, carrying out ultrasonic treatment for 10-15 min, preserving the solid phase micro-extraction device in the acetonitrile until the solid phase micro-extraction device is used after the ultrasonic treatment is finished, and taking out the solid phase micro-extraction device for air drying when the solid phase micro-extraction device is used.
Preferably, tetrahydrofuran is used for desorbing the adsorbed solid phase micro-extraction device in the step 3), the treatment time is 15-20 min, and ultrasonic treatment is accompanied in the desorption process.
Preferably, in the step 3), the water sample to be tested is simultaneously subjected to ultrasonic treatment and stirring treatment in the process of extracting the antibacterial agent by using the solid-phase micro-extraction device.
Preferably, the stirring speed is 800-820 rpm; the ultrasonic frequency is 20-30 kHz.
Preferably, the analysis and detection conditions of the high performance liquid chromatograph are as follows: the chromatographic column is a pentafluorophenyl column, and the mobile phase is a liquid phase with a volume ratio of 35: 35: 30 of mixed solution of methanol, acetonitrile and water, the flow rate is 1.0mL/min, the detection wavelength of a photodiode array detector is 211nm, and the sample injection volume is 20 mu L.
Preferably, the antimicrobial agent comprises one or more of TCS, TCC or M-TCS.
The invention also provides a solid phase micro-extraction device, which comprises an inner layer extraction membrane bag and an outer layer extraction membrane bag, wherein the inner layer extraction membrane bag is filled with graphene as an adsorbent; the outer extraction membrane bag is sleeved outside the inner extraction membrane bag, and a sinker is arranged in the outer extraction membrane bag and used for realizing the underwater positioning of the solid-phase micro-extraction device; the inner layer extraction film bag and the outer layer extraction film bag are prepared by hot-sealing polypropylene films on three sides.
3. Advantageous effects
Compared with the prior art, the invention has the beneficial effects that:
(1) according to the invention, the antibacterial agent in the water sample to be detected is extracted by adopting the solid-phase micro-extraction device, and then the antibacterial agent is desorbed and then is measured, so that the antibacterial agent can be effectively enriched and desorbed into the organic solvent, and the technical problem that the detection is inaccurate by adopting a high performance liquid chromatography when the antibacterial agent in the water sample is unevenly distributed or the content of the antibacterial agent is low is solved;
(2) the invention provides a detection method of an antibacterial agent in a water body based on graphene solid-phase extraction, wherein a solid-phase micro-extraction device is a micro solid-phase extraction device and comprises a double-layer extraction membrane bag, and graphene is filled in the inner layer extraction membrane bag and is used as an adsorbent; the sinker has been placed to outer extraction membrane bag, can realize the location of solid phase micro-extraction device under water for this equipment all remains in sample container's bottom in whole extraction process, avoids the extraction in-process, and miniature solid phase extraction device come-up is on the liquid surface or adhesion is on the container wall, has restricted the mixture of adsorbent and the water sample that awaits measuring, influences adsorption effect.
(3) The detection method of the antibacterial agent in the water body based on the graphene solid-phase extraction is provided with the inner-layer extraction membrane bag and the outer-layer extraction membrane bag, so that the adsorbent is effectively prevented from leaking into the water body to be detected;
the adsorbent is filled in the inner layer extraction membrane bag, the sinker is placed in the outer layer extraction membrane bag, and the two extraction membrane bags are arranged separately, so that the problem that the leakage of the adsorbent along a heat sealing area is aggravated due to the pushing effect of the movement of the sinker on the adsorbent can be effectively avoided;
(4) according to the method for detecting the antibacterial agent in the water body based on graphene solid-phase extraction, graphene is used as a solid-phase extraction adsorbent, and an HPLC method based on a pentafluorophenyl (PFP) column is combined, so that the contents of TCS, TCC and M-TCS in the water body can be accurately detected;
graphene has the following advantages as an adsorbent: the surface area is large, so that the load capacity is high, and both surfaces of the layer can absorb the waste heat; has good chemical stability and mechanical property; the interaction between graphene layers is strong, and the graphene layers have strong affinity with TCS, TCC and M-TCS containing benzene ring structures, strong adsorption effect and cheap synthetic raw materials;
in a high performance liquid chromatography test, a pentafluorophenyl (PFP) chromatographic column is selected, and due to the fact that fluorine groups in a PFP stationary phase are connected with polar urea groups through hydrogen bonds, the retention capacity of TCC is enhanced, and compared with chromatographic columns of C18, C8, amino and phenyl, the resolution is improved;
(5) according to the method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction, acetonitrile is selected to activate the solid-phase micro-extraction device, so that the diffusion of the analyte through the membrane is accelerated, and the acetonitrile has the effects of regulating the adsorbent and removing pollutants.
(6) According to the method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction, the pH value of a water sample to be detected is adjusted to 7.0-7.5 by using a phosphate buffer solution before extraction, so that protonation of urea groups of TCC (cross-type resistance) under an acidic condition is avoided, and the extraction effect of TCC can be effectively improved;
(7) according to the method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction, tetrahydrofuran is selected as an analytic solution, the tetrahydrofuran has proper polarity (0.207) to dissolve three analytes, and meanwhile, the tetrahydrofuran can be directly injected into an HPLC chromatographic column without affecting operation;
(8) according to the method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction, the solid-phase micro-extraction device is utilized to perform ultrasonic and stirring treatment on a water sample to be detected or a standard solution simultaneously in the process of extracting the antibacterial agent, so that the convection and the movement of the solution can be realized; the full contact between the adsorbent and the solution can be realized, and the aggregation of the adsorbent and the leakage from a packaging line can be prevented;
the simultaneous ultrasonic and stirring treatment not only avoids the problem that the total area of the mass transfer interface of the adsorbent cannot be increased when single magnetic stirring is used for assisting extraction, but also has limited improvement on the capacity of the total mass transfer rate; meanwhile, the problem that the extraction effect is influenced if the generated cavitation cannot ensure the maximization of the average concentration gradient of the system during single ultrasonic-assisted extraction is solved;
in the process of extracting the antibacterial agent by using a solid-phase microextraction device, the stirring speed needs to be controlled at 800-820 rpm, so that the adsorbent is ensured to achieve the optimal adsorption effect, and the extraction effect is ensured; the stirring speed is too high, so that the work of the shearing force generated by stirring on the cavitation nucleus is more than the work of the tensile stress for forming cavitation bubbles, and the cavitation is inhibited; meanwhile, too fast stirring speed can cause too large stirring vortex, increase the collision probability among small liquid drops of the solution, prevent the further reduction of the particle size of the small liquid drops and influence the extraction effect.
Drawings
FIG. 1 is a schematic structural diagram of a solid-phase microextraction device in accordance with the present invention;
FIG. 2 is a schematic view showing the structure of a solid-phase microextraction apparatus in comparative example 1-1;
FIG. 3 is a standard curve of target chromatographic peak area versus concentration; wherein: FIG. 3(a) is a TCS (0.5-1000. mu.g/L) standard curve (y: 0.0119x +0.0674, R)20.997); FIG. 3(b) is a TCC (0.2-1000 μ g/L) standard curve (y is 0.0073x +0.0732, R20.990); FIG. 3(c) is a standard curve of M-TCS (0.5-1000. mu.g/L) (y is 0.0089x +0.0747, R2＝0.996)；
FIG. 4 is a liquid chromatogram of TCS, TCC and M-TCS in example 1; a corresponds to the detection result of the sample with direct sample injection and standard addition (0.5 mug/L), and b corresponds to the detection result of the sample injection after the sample with standard addition is extracted by the solid phase micro-extraction device in the embodiment 1 of the invention.
In the figure: 1. an outer extraction membrane bag; 2. an inner layer extraction membrane bag; 3. an adsorbent; 4. and (5) sinking the seeds.
Detailed Description
It will be understood that when an element is referred to as being "mounted on" another element, it can be directly on the other element or the two elements can be directly connected together; when an element is referred to as being "connected" to another element, it can be directly connected to the other element or the two elements may be directly integrated. In addition, the terms "upper", "lower", "left", "right", "middle", "front" and "rear" used in the present specification are for convenience of description and are not intended to limit the scope of the present invention, and changes or modifications of the relative relationship between the terms and the terms are considered to be within the scope of the present invention without substantial changes in the technical contents.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs; as used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
Those not indicated in the examples, performed according to conventional conditions in the art or conditions suggested by the skilled person; the reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
As used herein, the terms "about," "near," and the like are also used for ease of description only to provide the flexibility associated with a given term, metric, or value. The degree of flexibility for a particular variable can be readily determined by one skilled in the art.
As used herein, "adjacent" refers to two structures or elements being in proximity. In particular, elements identified as "adjacent" may abut or be connected. Such elements may also be near or proximate to each other without necessarily contacting each other. In some cases, the precise degree of proximity may depend on the particular context.
Lengths, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such a range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a numerical range of about 1 to about 4.5 should be interpreted to include not only the explicitly recited limit values of 1 to about 4.5, but also include individual numbers (such as 2, 3, 4) and sub-ranges (such as 1 to 3, 2 to 4, etc.). The same principle applies to ranges reciting only one numerical value, such as "less than about 4.5," which should be construed to include all of the aforementioned values and ranges. Moreover, such an interpretation should apply regardless of the breadth of the range or feature being described.
The invention is further described below with reference to the accompanying drawings.
As shown in fig. 1, the solid-phase micro-extraction device in the present invention is a micro solid-phase micro-extraction device (hereinafter referred to as Mini-bar μ -SPE), the Mini-bar μ -SPE includes a double-layer extraction membrane bag, and the inner layer extraction membrane bag is filled with graphene as an adsorbent; sinkers are placed in the outer extraction membrane bag and used for realizing the underwater positioning of the solid-phase micro-extraction device; the sinker can be a small metal bar or other materials capable of realizing the underwater positioning of the Mini-bar mu-SPE.
The inner layer extraction film bag and the outer layer extraction film bag are both prepared by a polypropylene film through three-surface heat sealing.
Example 1
As shown in fig. 1, in the Mini-bar μ -SPE-1 of the present embodiment, the inner and outer extraction membrane bags are each prepared by heat sealing a polypropylene membrane on three sides, specifically:
inner layer extraction membrane bag: folding a polypropylene membrane, heating and sealing from two sides, then filling 20mg of adsorbent graphene, heating, sealing and sealing again, wherein the size of the inner layer extraction membrane bag is 1.5cm multiplied by 0.8 cm;
outer extraction membrane bag: folding a polypropylene membrane, heating and sealing from two sides, then filling the inner layer extraction membrane bag which is sealed and filled with the adsorbent, simultaneously filling a metal rod (diameter is 1 mm; length is 1.5cm), and finally, heat-sealing the opening end of the outer layer extraction membrane bag to fix the content, wherein the size of the outer layer extraction membrane bag is 2.0cm multiplied by 0.8 cm; and finishing mini-bar mu-SPE-1 assembly.
In this example, the conditions for the hplc assay were: the chromatographic column is a pentafluorophenyl column, and the mobile phase is a liquid phase with a volume ratio of 35: 35: 30 of mixed solution of methanol, acetonitrile and water, the flow rate is 1.0mL/min, the detection wavelength of a photodiode array detector is 211nm, and the sample injection volume is 20 mu L.
The method for detecting the antibacterial agent in the water body by using the Mini-bar mu-SPE-1 comprises the following specific steps:
1) pretreatment of Mini-bar μ -SPE-1: activating an adsorbent in a solid phase micro-extraction device by using acetonitrile; the method specifically comprises the following steps: soaking each Mini-bar mu-SPE-1 in acetonitrile, performing ultrasonic treatment for 10min, storing in the acetonitrile until the use after the ultrasonic treatment is finished, and taking out the solid phase micro-extraction device for air drying when in use;
2) standard solutions were prepared and standard curves plotted: dissolving TCS, TCC and M-TCS in acetonitrile to obtain corresponding stock solutions (10mg/L), adding water to dilute the stock solutions into seven standard solutions with concentration gradients, wherein the concentrations of the seven standard solutions containing TCS and M-TCS are respectively 0.5, 5, 10, 50, 100, 500 and 1000 mu g/L, and the concentrations of the seven standard solutions containing TCC are respectively 0.2, 2, 10, 20, 100, 500 and 1000 mu g/L, and analyzing and detecting the seven standard solutions by HPLC to obtain a linear relation of the concentration of a target compound to a chromatographic peak area response value, namely a calibrated standard curve;
the standard curve was prepared with the concentration of the target compound to be measured (μ g/L) as the abscissa and the measured value (peak area) as the ordinate. FIG. 3 shows a standard curve of the peak area of the target substance chromatogram versus the concentration; r2Values ranging from 0.990 to 0.997;
fig. 3 (a): the standard curve of TCS (0.5-1000 mu g/L) is as follows: 0.0119x +0.0674, R2A value of 0.997;
fig. 3 (b): the standard curve of TCC (0.2-1000 mu g/L) is as follows: y is 0.0073x +0.0732, R2A value of 0.990;
fig. 3 (c): the standard curve of M-TCS (0.5-1000 mu g/L) is as follows: y 0.0089x +0.0747, R2A value of 0.996;
3) pretreatment of a water sample to be detected and detection of the content of an antibacterial agent: in the embodiment, TCS, TCC and a solution with the concentration of M-TCS of 0.5 mug/L are prepared, a water sample to be detected is added, the collected water sample is filtered by Whatman filter paper and stored at 4 ℃ until further use;
before extraction, the pH value of a standard water sample to be detected is adjusted to 7 by using a phosphate buffer solution;
in the process of extracting the antibacterial agent by using the solid-phase microextraction device, simultaneously carrying out ultrasonic treatment and stirring treatment on a water sample to be detected, and additionally, placing a sinker in an outer extraction membrane bag of the micro solid-phase extraction device, so that the solid-phase microextraction device can be positioned underwater, graphene in the inner extraction membrane bag is in an irregular swinging motion state in the extraction process, and the swinging aggregation of the adsorbent towards the same fixed direction can be avoided, and on one hand, the adsorption effect can be prevented from being influenced by the aggregation of the adsorbent (especially for the adsorbent with larger specific surface area, such as graphene, if aggregation occurs in the adsorption process, the adsorption effect is inevitably influenced); on the other hand, the extraction membrane bag is prepared from a polypropylene membrane through three-side hot-sealing, so that leakage along a hot-sealing area due to adsorbent accumulation can be avoided (the leakage is mainly caused by adsorption of graphene on the polypropylene membrane wall, and the sealing quality is poor). The method for extracting the antibacterial agent in the standard water sample to be detected by using the solid phase microextraction device comprises the following specific steps: the Mini-bar μ -SPE-1 was immersed in a sample of spiked water (30mL) to be tested with magnetic stirring (800rpm) and sonication (20kHz), and after 2h the Mini-bar μ -SPE-1 device was removed, washed twice with water and dried with lint-free tissue. Then the Mini-bar mu-SPE-1 is put into THF (600 mu L), desorbed for 15 minutes under ultrasonic treatment, the extract (20 mu L) is directly introduced into HPLC for analysis and detection, the obtained measured value is calculated by contrasting with a standard curve to obtain the contents of TCS, TCC and M-TCS in the standard water sample of 0.67 mu g/L, 0.19 mu g/L and 0.44 mu g/L respectively, and the recovery rates are 134%, 38% and 88% respectively, as shown in figure 4 b.
The measurement results (peak area expression) are shown in table 1:
TABLE 1 Peak area of target Compound/target Compound content
Figure BDA0002547337200000071
For comparison, after the water sample to be detected is filtered by filter paper, sample introduction is directly performed to HPLC, and the chromatogram in FIG. 4a is obtained. Due to the uneven distribution of the antibacterial agent and the low content of the antibacterial agent in the water sample, only the chromatographic peak of TCS appears in FIG. 4 a; as can be seen from the chromatogram of fig. 4b obtained after extraction and desorption by the solid-phase microextraction device in this example, the enrichment capacity of the method of the present invention is strong, and shows the high resolution of the three compounds separated from the matrix component at low concentration.
Comparative examples 1 to 1
Mini-bar μ -SPE-2 in this comparative example: the Mini-bar mu-SPE-2 device is essentially the same as example 1 except that the metal rod and the graphene adsorbent are both encapsulated in an inner extraction membrane bag (as shown in fig. 2).
In this comparative example, the measurement conditions of high performance liquid chromatography were the same as in example 1.
In the comparative example, solutions with the concentrations of TCS, TCC and M-TCS of 100 mug/L are prepared as water bodies to be tested, and the following are respectively utilized: detecting the micro-bar mu-SPE-1 and the micro-bar mu-SPE-2;
and (3) respectively utilizing the two Mini-bar mu-SPEs to detect the antibacterial agent in the water body, and specifically comprising the following steps:
1) pretreatment of Mini-bar μ -SPE: the same as example 1;
2) before extraction, the pH value of a water sample to be detected is adjusted to 7.0 by using a phosphate buffer solution;
extracting the antibacterial agent in the water sample to be detected by using a solid phase microextraction device; specifically, the Mini-bar μ -SPE was dipped into standard solution (30mL) with magnetic stirring (800rpm) and sonication (20kHz), and after 2h, the Mini-bar μ -SPE unit was removed, washed twice with water and dried with lint-free tissue. Then Mini-bar μ -SPE was placed in THF (600 μ L), desorbed for 15 minutes under sonication, the extract (20 μ L) was directly introduced into HPLC for analytical detection, and the obtained measurements were calculated against a standard curve: the contents of TCS, TCC and M-TCS in the water sample after being extracted by a Mini-bar mu-SPE-1 solid phase micro-extraction device are respectively 96.23 mu g/L, 95.36 mu g/L and 98.64 mu g/L; the contents of TCS, TCC and M-TCS in the water sample after being extracted by the Mini-bar mu-SPE-2 solid phase micro-extraction device are respectively 78.44 mu g/L, 80.61 mu g/L and 83.29 mu g/L.
The measurement results (peak area expression) are shown in table 2:
TABLE 2 Peak area of target Compound/target Compound content
Figure BDA0002547337200000081
The results show that compared with the Mini-bar mu-SPE-2 solid phase micro-extraction device, the Mini-bar mu-SPE-1 solid phase micro-extraction device has better extraction effect, and the recovery rate of TCS, TCC and M-TCS is obviously higher than that of the Mini-bar mu-SPE-2 solid phase micro-extraction device.
Comparative examples 1 to 2
The Mini-bar μ -SPE device in this comparative example is the same as the Mini-bar μ -SPE-1 device used in comparative example 1-1 (shown in FIG. 1).
In this comparative example, the HPLC measurement conditions were the same as those of the Mini-bar μ -SPE-1 apparatus used in comparative example 1-1.
In the comparative example, six parts of the same TCS, TCC and solution (C1-C6) with the concentration of M-TCS being 100 mug/L are prepared and respectively detected by utilizing Mini-bar mu-SPE-1;
the detection of the antibacterial agent in the water body is carried out by utilizing the Mini-bar mu-SPE-1, and the specific steps are different from those of the comparative example 1-1 only in that:
when solution C1 was extracted: in the step 2), only performing magnetic stirring (800rpm) treatment when extracting the antibacterial agent in the water sample to be detected by using a solid phase microextraction device;
when solution C2 was extracted: in the step 2), only ultrasonic (20kHz) treatment is carried out when the solid-phase microextraction device is used for extracting the antibacterial agent in the water sample to be detected;
when solution C3 was extracted: in the step 2), magnetic stirring (600rpm) treatment is carried out when the solid-phase microextraction device is used for extracting the antibacterial agent in the water sample to be detected, and ultrasonic treatment (20kHz) is carried out simultaneously;
when solution C4 was extracted: in the step 2), magnetic stirring (1000rpm) treatment is carried out when the solid-phase microextraction device is used for extracting the antibacterial agent in the water sample to be detected, and ultrasonic treatment (20kHz) is carried out simultaneously;
when solution C5 was extracted: in the step 2), magnetic stirring (800rpm) treatment is carried out when the solid-phase microextraction device is used for extracting the antibacterial agent in the water sample to be detected, and ultrasonic treatment (40kHz) is carried out simultaneously;
when solution C6 was extracted: in the step 2), magnetic stirring (800rpm) treatment is carried out when the solid-phase microextraction device is used for extracting the antibacterial agent in the water sample to be detected, and ultrasonic treatment (50kHz) is carried out simultaneously;
the measurement results (peak area expression) are shown in table 3:
TABLE 3 Peak area of target Compound/target Compound content
Figure BDA0002547337200000091
The results show that when the Mini-bar mu-SPE solid phase micro-extraction device is used for extracting the antibacterial agent in the water sample to be detected, the extraction effect is optimal after the magnetic stirring (800rpm) treatment and the ultrasonic (20kHz) treatment are carried out simultaneously.
Example 2
As a blank labeling experiment of example 1, this example is different from example 1 only in that: and replacing the water sample to be detected with distilled water, similarly preparing a solution with the concentration of TCS, TCC and M-TCS of 0.5 mu g/L, adding distilled water, extracting by a Mini-bar mu-SPE-1 solid phase micro-extraction device to obtain a standard water sample containing TCS, TCC and M-TCS of 0.41 mu g/L, 0.12 mu g/L and 0.39 mu g/L respectively, and recovering the recovery rates of 82%, 24% and 78% respectively.
Example 3
This example differs from example 1 only in that: in the step 3), the magnetic stirring speeds of the solid-phase microextraction device for extracting the antibacterial agent in the water sample to be detected are 810rpm and 820rpm respectively, the same water sample to be detected is treated by the solid-phase microextraction device in the same way as in the example 1, and the obtained extraction result of the target compound is basically the same as that in the example 1.
Example 4
This example differs from example 1 only in that: when the solid-phase microextraction device is used for extracting the antibacterial agent in the water sample to be detected in the step 3), the ultrasonic frequency is respectively 25kHz and 30kHz, the water sample to be detected is treated the same as that in the example 1, and the obtained extraction result of the target compound is basically the same as that in the example 1.
The above examples are only for illustrating the technical idea and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the content of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.

Claims (10)

1. A detection method of an antibacterial agent in a water body based on graphene solid-phase extraction is characterized by comprising the following steps:
1) pretreatment of a solid phase micro-extraction device: activating an adsorbent in a solid phase micro-extraction device by using acetonitrile;
2) preparing an antibacterial agent standard solution, measuring by using a high performance liquid chromatograph, and drawing a standard curve;
3) pretreatment of a water sample to be detected and detection of the content of an antibacterial agent: filtering a water sample to be detected, and adjusting the pH value of the water sample to be detected to 7.0-7.5;
extracting the antibacterial agent in the water sample to be detected by using a solid phase microextraction device;
and (3) carrying out desorption treatment on the adsorbed solid-phase micro-extraction device, and injecting the desorption solution into a high performance liquid chromatograph for analysis and detection to obtain the content of the antibacterial agent in the water sample.
2. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction according to claim 1, wherein the solid-phase micro-extraction device is a micro solid-phase extraction device and comprises an inner-layer extraction membrane bag and an outer-layer extraction membrane bag, and the inner-layer extraction membrane bag is filled with graphene serving as an adsorbent; the outer extraction membrane bag is sleeved outside the inner extraction membrane bag, and a sinker is arranged in the outer extraction membrane bag and used for realizing the underwater positioning of the solid-phase micro-extraction device; the inner layer extraction film bag and the outer layer extraction film bag are prepared by hot-sealing polypropylene films on three sides.
3. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction as claimed in claim 2, wherein the relationship between the amount of graphene and the antibacterial agent content in the water sample to be extracted is as follows: when the concentration of the antibacterial agent is less than or equal to 1000 mug/L, 20mg of graphene is filled in the solid-phase micro-extraction device.
4. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction according to claim 2, wherein the activation treatment in the step 1) is specifically: and (3) soaking the solid phase micro-extraction device in acetonitrile, carrying out ultrasonic treatment for 10-15 min, preserving the solid phase micro-extraction device in the acetonitrile until the solid phase micro-extraction device is used after the ultrasonic treatment is finished, and taking out the solid phase micro-extraction device for air drying when the solid phase micro-extraction device is used.
5. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction according to claim 2, wherein tetrahydrofuran is used for desorbing the adsorbed solid-phase microextraction device in the step 3), the treatment time is 15-20 min, and ultrasonic treatment is accompanied in the desorption process.
6. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction according to claim 2, wherein in the step 3), the antibacterial agent is extracted by using a solid-phase micro-extraction device, and a water sample to be detected is subjected to ultrasonic treatment and stirring treatment simultaneously.
7. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction is characterized in that the stirring speed is 800-820 rpm; the ultrasonic frequency is 20-30 kHz.
8. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction according to any one of claims 1 to 7, wherein the analysis and detection conditions of the high performance liquid chromatograph are as follows: the chromatographic column is a pentafluorophenyl column, and the mobile phase is a liquid phase with a volume ratio of 35: 35: 30 of mixed solution of methanol, acetonitrile and water, the flow rate is 1.0mL/min, the detection wavelength of a photodiode array detector is 211nm, and the sample injection volume is 20 mu L.
9. The method for detecting the antibacterial agent in the water body based on the graphene solid-phase extraction according to claim 8, wherein the antibacterial agent comprises one or more of TCS, TCC or M-TCS.
10. The solid-phase micro-extraction device is characterized by comprising an inner-layer extraction membrane bag and an outer-layer extraction membrane bag, wherein the inner-layer extraction membrane bag is filled with graphene serving as an adsorbent; the outer extraction membrane bag is sleeved outside the inner extraction membrane bag, and a sinker is arranged in the outer extraction membrane bag and used for realizing the underwater positioning of the solid-phase micro-extraction device; the inner layer extraction film bag and the outer layer extraction film bag are prepared by hot-sealing polypropylene films on three sides.
CN202010564997.9A 2020-06-19 2020-06-19 Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device Pending CN111595989A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202010564997.9A CN111595989A (en) 2020-06-19 2020-06-19 Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202010564997.9A CN111595989A (en) 2020-06-19 2020-06-19 Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device

Publications (1)

Publication Number Publication Date
CN111595989A true CN111595989A (en) 2020-08-28

Family

ID=72184837

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202010564997.9A Pending CN111595989A (en) 2020-06-19 2020-06-19 Graphene solid-phase extraction-based detection method for antibacterial agent in water body and solid-phase micro-extraction device

Country Status (1)

Country Link
CN (1) CN111595989A (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103728394A (en) * 2014-01-15 2014-04-16 中华人民共和国苏州出入境检验检疫局 Daily chemical product antibacterial agent detection method based on graphene oxide solid phase extraction
CN207221389U (en) * 2017-08-18 2018-04-13 河海大学 A kind of micro- solid-phase extraction device
CN109932449A (en) * 2019-04-03 2019-06-25 东北师范大学 A kind of preparation of magnetic porous graphene and its rapid detection method applied to triclosan at low triclosan concentrations in water

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103728394A (en) * 2014-01-15 2014-04-16 中华人民共和国苏州出入境检验检疫局 Daily chemical product antibacterial agent detection method based on graphene oxide solid phase extraction
CN207221389U (en) * 2017-08-18 2018-04-13 河海大学 A kind of micro- solid-phase extraction device
CN109932449A (en) * 2019-04-03 2019-06-25 东北师范大学 A kind of preparation of magnetic porous graphene and its rapid detection method applied to triclosan at low triclosan concentrations in water

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ANAS ALSHISHANI ET AL.: "High performance liquid chromatographic determination of triclosan, triclocarban and methyl-triclosan in wastewater using mini-bar micro-solid phase extraction", 《MICROCHEMICAL JOURNAL》 *

Similar Documents

Publication Publication Date Title
Mei et al. Determination of trace bisphenol A in complex samples using selective molecularly imprinted solid-phase extraction coupled with capillary electrophoresis
Roldán-Pijuán et al. Stir fabric phase sorptive extraction for the determination of triazine herbicides in environmental waters by liquid chromatography
Zhao et al. Preparation of dummy template imprinted polymers at surface of silica microparticles for the selective extraction of trace bisphenol A from water samples
Wu et al. A novel surface molecularly imprinted polymer as the solid-phase extraction adsorbent for the selective determination of ampicillin sodium in milk and blood samples
Mei et al. Sensitive monitoring of benzoylurea insecticides in water and juice samples treated with multiple monolithic fiber solid-phase microextraction and liquid chromatographic analysis
Kueseng et al. Multiwalled carbon nanotubes/cryogel composite, a new sorbent for determination of trace polycyclic aromatic hydrocarbons
CN104558361B (en) Preparation method and application of hydrophilic polyaromatic hydrocarbon molecularly imprinted solid phase extraction filler
Huang et al. Preparation and application of ionic liquid-coated fused-silica capillary fibers for solid-phase microextraction
Chen et al. A graphene oxide surface–molecularly imprinted polymer as a dispersive solid-phase extraction adsorbent for the determination of cefadroxil in water samples
He et al. Determination of diethylstilbestrol in seawater by molecularly imprinted solid-phase extraction coupled with high-performance liquid chromatography
CN104151489A (en) Hydrophilic hydroxyl polychlorinated biphenyl molecularly imprinted solid-phase extraction filler as well as preparation method and application thereof
Du et al. Selective extraction of dimethoate from cucumber samples by use of molecularly imprinted microspheres
Zhang et al. Porous molecularly imprinted monolithic capillary column for on-line extraction coupled to high-performance liquid chromatography for trace analysis of antimicrobials in food samples
Jin et al. Synthesis and evaluation of molecularly imprinted polymer for the determination of the phthalate esters in the bottled beverages by HPLC
Zhao et al. Determination of carbamate pesticides in water and fruit samples using carbon nanotube reinforced hollow fiber liquid-phase microextraction followed by high performance liquid chromatography
Huang et al. Fabric phase sorptive extraction: Two practical sample pretreatment techniques for brominated flame retardants in water
Zhou et al. Comparison of multiwalled carbon nanotubes and a conventional absorbent on the enrichment of sulfonylurea herbicides in water samples
Jorn et al. Determination of ochratoxin A in red wines by multiple pulsed elutions from molecularly imprinted polypyrrole
Cui et al. Molecularly imprinted electrospun nanofibre membrane assisted stir bar sorptive extraction for trace analysis of sulfonamides from animal feeds
Liu et al. Removal of persistent organic pollutants from micro-polluted drinking water by triolein embedded absorbent
Lian et al. Selective extraction and concentration of mebendazole in seawater samples using molecularly imprinted polymer as sorbent
Chu et al. Packed-nanofiber solid phase extraction coupled with HPLC for the determination of chloramphenicol in milk
CN111239268B (en) Method for determining amide compounds in environmental water sample by solid-phase extraction-liquid chromatography
CN106168607B (en) A method of utilizing phthalate ester in solid phase microextraction and gas chromatography combined with mass spectrometry technology detection water body
Liu et al. Migration detection of six aromatic amines in polyamide food contact materials by HPLC after molecularly imprinted polymer pipette tip solid phase extraction

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20200828