Green preparation of chlorine-doped graphene and its application in electrochemical sensor for chloramphenicol detection
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A green but efficient synthesis approach for chlorine-doped reduced graphene oxide (Cl-RGO) was developed. The chlorine-doping and reduction was achieved in one-step by refluxing graphene oxide solution in concentrated hydrochloric acid (8 M, 120 °C) under N2 atmosphere. X-ray photoelectron spectroscopy analysis revealed that the Cl content was 1.01 at.% in the Cl-RGO. Electrochemical measurements indicated that the Cl-RGO modified glass carbon electrode showed enhanced electrochemical conductivity and electrocatalytic activity for the veterinary drug chloramphenicol (CAP) detection. Thus a highly selective electrochemical sensor for CAP was constructed based on Cl-RGO, and a linear relationship between current intensity and CAP concentration (2–35 μM) was obtained with a detection limit of 1 μM (S/N = 3). The sensor showed excellent reproducibility, storage stability and was successfully used for CAP detection in milk, calf plasma, water and pharmaceutical samples.
KeywordsChlorine-doped graphene Electrochemical sensor Chloramphenicol Green synthesis
Chloramphenicol (CAP) is a broad-spectrum antibacterial veterinary drug and has been extensively used for the treatment of infectious diseases in animals. However, it has been recognized that the CAP may cause many chronic diseases such as bone marrow depression, aplastic anemia and cardiovascular collapse [1, 2, 3]. As a consequence, the use of CAP in animals-derived food has been globally banned to control the food safety. The increasing concerns about food safety urged the development of rapid, selective and sensitive analytical methods to monitor CAP residue in food and water samples.
Nowadays, various conventional analytical methods, such as liquid chromatography-mass spectroscopy (LC–MS) , gas chromatography-mass spectroscopy (GC–MS) , capillary electrophoresis (CE) , and chemiluminescent immunoassay , have been put forward for CAP residue detection. Nevertheless, complicated pretreatment processes, expensive instruments and professional operator requirements make these methods not suitable to routine and rapid analysis of samples. Alternatively, electrochemical method has the advantages of rapid analysis, cheap equipment and simple operation [8, 9]. Thus many electrochemical sensors for CAP detection based on various electrode materials have been developed [10, 11, 12, 13, 14, 15, 16, 17]. Among these electrode materials, various nanomaterials such as carbon and metal nanoparticles, have been employed for antibiotics drug residues detection in animal-derived food and water samples [18, 19, 20, 21]. As a typical carbon nanomaterial, graphene has drawn increasing interest as electrode materials because of its excellent electrical conductivity, high surface area and mechanical strength . Recently, graphene-based electrochemical sensors for CAP have been developed, for example, titanium nitride–reduced graphene oxide nanohybrids , gold nanoparticles/nitrogen-doped graphene , silver nanoparticles/graphene composite [24, 25]. Most of these reported graphene-based electrode materials require complicated preparation processes and the electrochemical sensing performances should be further improved.
As well-known, heteroatom-doping was an effective approach to tailor the electrochemical properties of graphene materials [26, 27, 28]. In this regard, nitrogen-, sulfur-, phosphorus- and boron-doped graphene materials have been facilely derived from chemical reduction of graphene oxide (GO) and used as new electrode materials [28, 29, 30]. The preparation of heteroatom-doped graphene usually needs harsh experimental condition and toxic chemical regent. In this work, a green and efficient synthesis approach for chlorine-doped reduced graphene oxide (Cl-RGO) was developed. The chlorine-doping and reduction was achieved in one-step by refluxing GO solution in concentrated hydrochloric (HCl) acid under N2 atmosphere. The Cl-RGO was further used as a novel electrode material to construct electrochemical sensor for veterinary drug CAP detection in milk, calf plasma, water and pharmaceutical samples.
All common chemicals were obtained from Sinopharm Chemical Reagent Corp. (Shanghai, China) at analytical grade. Graphite powder (99.85%) was purchased from XFNANO (Nanjing, China). CAP was supplied by Hefei Bomei Biotechnology Co. Ltd. (China). Phosphate buffer solutions (PBS, 0.1 M) were prepared by mixing K2HPO4 and KH2PO4 and used for CAP stock solution (3 mM) preparation and electrochemical measurements.
2.2 Preparation of Cl-RGO
GO was firstly produced from graphite powder according to the previously reported procedures [31, 32]. Then the GO (20 mg) was dispersed in water (13 mL) and concentrated hydrochloric acid (27 mL) under ultrasonication for 30 min. The GO solution was transferred into a round bottom flask and heated in oil bath at 120 °C for 6 h under nitrogen atmosphere. After cooling to room temperature, the Cl-RGO was collected by centrifugation, thoroughly washing with water until neutral pH reached, and finally dried in vacuum oven at 45 °C for 12 h.
2.3 Characterization apparatus
The morphology Cl-RGO was characterized by scanning electron microscopy (SEM, Hitachi S-4800, Japan) and transmission electron microscopy (TEM, JEOL JEM-2100F, Japan). X-ray photoelectron spectroscopy (XPS) was recorded on PHI 5400 (USA).
2.4 Electrochemical measurements
The electrochemical measurements were performed as previously described procedures . In brief, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) measurements were performed on Chenhua CHI-660D electrochemical workstation (Shanghai, China) with a standard three-electrode system. Pre-polished glassy carbon electrode (GCE) was coated by Cl-RGO suspension and used as the working electrode (Cl-RGO/GCE), while Pt wire and saturated calomel electrode (SCE) were employed as counter as well as reference electrodes, respectively. The CV measurements were carried out at a potential range between − 0.2 and 0.6 V (vs. SCE) for [Fe(CN)6]3−/4− or − 0.8 to 0.4 V (vs. SCE) for CAP with the scan rate of 50 mV s−1. The DPV measurements was finished at a potential window between − 0.4 and − 0.8 V (vs. SCE), with step potential of 4 mV, amplitude of 50 mV, pulse width of 0.2 s and pulse period of 0.5 s, respectively.
3 Results and discussion
3.1 Optimization of preparation conditions for Cl-RGO
3.2 Characterization of Cl-RGO
3.3 Optimization of determination conditions for CAP
3.4 Analytical performances of CAP sensor
3.5 CAP detection in real samples
Determination of CAP in real samples
5.12 ± 0.3
10.35 ± 0.5
5.21 ± 0.2
20.31 ± 0.9
25.55a ± 0.3
29.98 ± 0.4
10.50 ± 0.1
19.86 ± 0.2
In summary, chlorine-doped reduced graphene oxide (Cl-RGO) was greenly fabricated and used as novel electrode material for electrochemical sensor. The efficient chlorine-doping (1.01 at.%) and reduction was achieved in one-step by refluxing graphene oxide (GO) solution in concentrated hydrochloric acid under N2 atmosphere. The electrochemical sensor for veterinary drug chloramphenicol (CAP) detection was constructed based on Cl-RGO coating on glass carbon electrode (GCE) with a detection limit of 1 μM (S/N = 3). Furthermore, the sensor showed excellent anti-interference ability, reproducibility, stability, and was successfully used for determination of CAP in milk, calf plasma, water and pharmaceutical samples with satisfactory recovery result. The simple and green preparation procedure as well as excellent electrocatalytic ability of Cl-RGO/GCE would be benefit for developing of electrochemical sensor for hazardous antibiotics detection in food safety testing.
This study was funded by Shanghai Municipal Natural Science Foundation (16ZR1401700) and the Open Research Fund of Key Laboratory of Polar Materials and Devices, Ministry of Education.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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