Original Article

Emerging techniques in food science: the resistance of chlorpyrifos pesticide pollution against arc and dielectric barrier discharge plasma

Mohsen Gavahian1*, Tsai Meng-Jen2, Amin Mousavi Khaneghah3*

1Department of Food Science, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan, ROC;

2Product and Process Research Center, Food Industry Research and Development Institute, Hsinchu, 30062, Taiwan, ROC;

3Department of Food Science, University of Campinas, Campinas – SP, Brazil


Many studies introduced cold plasma as a novel and effective processing technology for microbial decontamination of food and water as well as for the removal of environmental pollution such as pesticide. However, as there are several types of plasma designs, their efficacy in degrading major pesticide residues, such as chlorpyrifos (as a hazardous chemical), should be explored. This study was conducted to assess the decontamination efficacy of 8 min of arc and dielectric barrier discharge (DBD) plasma on chlorpyrifos pesticide-water samples at a concentration of 2 mg•L-1. The plasma-treated samples were assessed by liquid chromatography-mass spectrometry (LC-MS) and compared with the control (untreated) sample. In addition, the effects of plasma processes on some physical properties of samples were studied. According to the results, plasma-treated samples showed similar physical characteristics (e.g., refractive index and color values) to those of the untreated samples. While the temperature of the samples remained steady during the DBD plasma treatment, arc plasma changed the temperature of the sample at a rate of about 3.75°C•min–1 and yielded a sample with a final temperature of 60°C. However, contrary to the general belief that plasma is an efficient technique for pesticide degradation, chemical analyses showed high resistance of chlorpyrifos against both arc and DBD plasma under the conditions used in the present study. Therefore, the possibility of high resistance of pesticide pollution to this emerging technology should be considered. Also, further studies on the efficiency of the selected plasma system for removing pesticide pollution (e.g., during water and wastewater treatment) at industrial scale is needed.

Key words: decontamination, emerging processing technology, hazardous chemical, nonthermal plasma, pesticide pollution, pesticide reduction

Corresponding Authors: Mohsen Gavahian, Ph.D., Department of Food Science, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan, ROC. Email: [email protected];

Amin Mousavi Khaneghah, Ph.D., Department of Food Science, Faculty of Food Engineering, University of Campinas (UNICAMP), Rua Monteiro Lobato, 80. Caixa Postal: 6121.CEP: 13083-862, Campinas, São Paulo, Brazil. Email: [email protected]

Received: 01 September 2020; Accepted: 28 September 2020; Published: 14 October 2020

DOI: 10.15586/qas.v12iSP1.807

© 2020 Codon Publications

This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0). License (


The agricultural industry utilizes the approaches that stabilize the production rate and avoid economic loss unpleasant phenomena such as pest attack (Halimatunsadiah et al., 2016). For example, many farmers may use approved chemical pesticides in accordance with regulations (Guo et al., 2020; Heshmati et al., 2020). However, many environmental scientists warned about the negative effects of these contaminants because they act as environmental pollutants that pose negative health effects (Albasher et al., 2020; Ali et al., 2020). Food scientists also warned about the safety aspects of the agricultural products that contain pesticide residues (Serefoglu and Serefoglu, 2016). For instance, chlorpyrifos (O, O-diethyl-O-3,5,6-trichlor2-pyridyl phosphorothioate) has been widely employed in the agricultural industry for controlling a variety of crop pests, including white-ants and foliar insects, for more than half a century. According to the United States Environmental Protection Agency (EPA), direct food exposure of this pesticide for children is about 0.24 (μg•kg-1•day-1) (EPA, 2016). Also, a tolerance level of 100 ppb is defined for chlorpyrifos residues, per se, in or on food commodities (Regulations, 2020). It was revealed that consumption of the residues of this chemical can result in diarrhea, frequent urination, abnormal sweating, salivation, lacrimation, genotoxicity, neurochemical changes, neurobehavioral disorder, and abnormal birth outcomes (Anderson et al., 2018; Farkhondeh et al., 2020; Foong et al., 2020). Therefore, its indoor application was banned by EPA since the early 21st century. However, it is still among the most widely used and approved pesticides in the agriculture industry, with an annual consumption of about 4,000,000 kg in the United States (EPA, 2011a, 2011b). This health-threatening pesticide is also authorized for agricultural applications in many countries (Foong et al., 2020; Razzaghi et al., 2018).

Besides, food regulations can vary across regions, and insufficient attention of regulatory bodies in some developing countries may result in agricultural products that cannot comply with the food safety regulations of the importing country (e.g., due to high concentrations of pesticide residue). Therefore, the presence of chemical pesticide residues is challenging the agro-food industry. In this regard, scientists tried to develop effective approaches for the production of pesticide-free agricultural and food products by exploring the efficacy of decontamination techniques such as photocatalysis (Vigneshwaran et al., 2019) and ultrasound treatment (Jawale and Gogate, 2016), and ozone (Pandiselvam et al., 2020). However, none of the proposed methods satisfied the industry due to the production of unwanted by-products or low efficiency. Recently, cold plasma, which is a combination of electrons, photons, ions, free radicals, and other reactive species, has been introduced to the food industry, and it showed several potential applications, including pesticide degradation (Gavahian and Cullen, 2020; Gavahian and Khaneghah, 2020; Sarangapani et al., 2016). Besides, sustainability and sustainable development get attention from various industries (e.g., food and agriculture industry) in many regions of the world. For example, the central ministries of Taiwan, including EPA, Council of Agriculture (COA), and the Ministry of Economic Affairs (MOEA), have supported the regulatory promotion. In agreement with the sustainable development goals (SDGs) launched in Taiwan, it seems that cold plasma, specifically the application of this green technology for the sustainable decontamination of food, water, and wastewater, has the capacity to help in meeting the SDGs. A wide range of designs has been proposed for plasma applications in the food industry, such as arc, dielectric barrier discharge (DBD), glow, in-pack, and corona discharge, which necessitates exploring their performance in the production of pesticide-free products. Many studies introduced plasma as a promising tool for pesticide degradations. However, reports on the possible resistance of pesticides against plasma treatments are rare. Therefore, the present study aims to assess the feasibility of decontamination of chlorpyrifos, as a major pesticide residue in agricultural products, through DBD and arc plasma treatments.

Material and Methods

Figure 1 represents a summary of the methods used in the present study.

Figure 1. A graphical abstract representing the methods used in this study.


All the reagents were analytical grade and were obtained from Sigma–Aldrich (Missouri, United States). CAS 2921-88-2 (Chlorpyrifos pesticide) at a purity of 40.8 (w/w)% was procured from Chia-yi Chemical Industry Corporation (Tainan, Taiwan).

Pesticide preparation

A mixture of pesticide (as described in the “chemicals section”) in methanol prepared at 0.5 mg•mL–1 was diluted with distilled water to the concentration of 2 mg•L-1 (Sarangapani et al., 2016). The mixture was subjected to an electromagnetic stirrer to ensure that the pesticide is distributed thoroughly within the liquid phase.

Plasma equipment and process parameters

DBD and arc plasma devices were employed in the present work (Figure 2). Table 1 represents the process conditions of these plasma systems. For DBD plasma treatment, AAA-168 DBD plasma (APPL, Taiwan) was employed at a constant temperature of 30°C, which used nitrogen purity of 99.99% (Chiah Lung Enterprise Co., Taiwan) as the carrier gas. The pesticide sample (20 mL) was poured in a glass Petri dish (100 × 15 mm) to form a thin layer. It was then placed in the discharging area of the DBD plasma. More details about the DBD system used in the present study can be found in a previous publication (Gavahian et al., 2020). Likewise, the same amount of pesticide sample was subjected to arc atmospheric-pressure plasma (APPL-10k, Taiwan) with a working distance of 5 cm. More details about the arc plasma system used in the present study can be found in a previous publication (Gavahian et al., 2019b). The variations in the temperature of the sample during arc and DBD treatments were monitored through the TES 1306 thermometer (TES Electric Electronic Corp., Taiwan) and matching software (APPL, Taiwan), respectively. Both DBD and arc plasma treatments were applied for 8 min at a power of 0.3 kW in triplicate. This processing time was selected according to our preliminary study. Long processing times (e.g., 10 min) resulted in high temperatures of the sample in the arc system, which may degrade the pesticide, and such a degradation may be a reason for the misinterpretation of degradation data. The DBD-treated, arc-treated, and control (not treated by plasma) samples were kept in a dark and cool place before further studies.

Figure 2. The schematic representation of the plasma devices employed in the present investigation: arc plasma (a) and dielectric barrier discharge (DBD) plasma (b).

Table 1. Arc and DBD plasma process conditions in this study.

DBD* plasma Arc plasma
Input gas Nitrogen Atmospheric air
Pressure (kPa) 10 101
Process duration (min) 8 8
Temperature (°C) 30 30~60
Input power (W) 300 300

*DBD, dielectric barrier discharge.

Qualitative analysis of the pesticide residue

The chemical composition of the samples was explored by liquid chromatography-mass spectrometry (LC-MS) to identify the possibly degraded products of pesticide and the concentration of the remaining pesticide in the plasma-treated samples in comparison with the control sample according to the procedure that was previously explained by Shoeibi et al. (2013). Briefly, 5 g of each pesticide sample, that is, control, arc plasma-treated, or DBD plasma-treated, was mixed with distilled water (10 mL) and acetonitrile (10 mL) in a centrifuge tube with the capacity of 50 mL. Afterward, sodium chloride (1 g) and magnesium sulfate anhydrous (4 g) were added, which were vigorously shaken manually for 1 min before being centrifuged at 3500 rpm for 10 min. The supernatant was collected and passed through a nylon syringe filter that had a mesh size of 0.22 μm. The filtered liquid was subjected to LC-MS investigations. Hypersil Gold C-18 aQ (Thermo Scientific, USA) with the dimensions of 100 × 2.1 mm and an internal diameter of 2.6 μm was employed as the analytical column. Two mobile phases, that is, A and B, were used in the present work. The first one (mobile phase A) consists of ammonium formate (AF) with a concentration of 5 mM and 0.1% formic acid (FA) in the water while the latter (mobile phase B) contained AF 5 mM and 0.1% FA in methyl alcohol. The gradient elution program was as follows: 0–0.5 min (2% B), 0.5–2 min (2–40% B), 2–20 min (40–95% B), 20–22 min (at 95% B), 22.0–22.1 min (returned to 2% B), and allowed for equilibration for 180 s before the next injection. A constant flow rate of 300 μL.min-1 was used in the LC assessment. For HESI-MS, pure nitrogen was used as both auxiliary and sheath gas. The spray voltage of positive ionization mode was set at 3.5 kV. In addition, the temperatures of the capillary and auxiliary gas heaters were 320 and 350°C, respectively. The flow rate of the sheath and auxiliary gas were 40 and 10, respectively. Data collection was performed over an m/z range of 100 to 1000. Data independent was conducted at the normalized collision energy of 30 and 80 eV. The version 3.3 of TraceFinder (Thermo Scientific, USA) was used for processing the results.

Refractive index evaluation

A laboratory 0.0–33.0% Master-M Refractometer (ATAGO, Japan) along with the conversion table was employed for reflective index measurements. In this regard, samples were shaken, and 1 mL of the sample was placed on the daylight plate of the refractometer to obtain the results.

Color of the pesticide

The color of the pesticide was assessed through a CM-5 colorimeter (Konica Minolta Sensing, Japan) to obtain L*, a*, and b* values, which represent the lightness, greenness-redness, and blueness-yellowness of the sample, respectively (Gavahian et al., 2012). In addition, the variations in the overall color of the plasma-treated samples in comparison with the control sample were calculated through Eq. (1) (Gavahian et al., 2019a).

ΔE=(Lp*Lc*)2+(aP*ac*)2+(bP*bc*),      (1)

where L*p, a*p, and b*p are the lightness, greenness-redness, and blueness-yellowness values of plasma-treated samples, and L*c, a*c, and b*c are those of the control (untreated) sample.

Statistical analysis

The statistical analysis was performed using version 23 of the SPSS Statistics (IBM, USA). In this regard, the ANOVA along with the Post Hoc-Duncan assay at a 0.95 confidence interval was used to detect significant differences between the mean values.

Result and discussion

Liquid chromatography-mass spectrometry of pesticide residue

Figure 3 represents the LC-MS chromatograms of control and plasma-treated samples. According to the data, a similar peak was identified at a retention time of 17.8 min for all the samples. These peaks have a similar intensity of about 4 × 108. Besides, no additional peaks were detected in either DBD or arc plasma-treated samples during 20 min of elution time. Therefore, neither arc nor DBD plasma process degraded the chlorpyrifos, and this pesticide resisted against these processes. It was expected that the plasma-generated reactive species, such as free radicals, ozone, and ions, affect the chemical structure of the chlorpyrifos. However, for the first time, the present investigation disclosed that this pesticide remained unchanged after plasma treatment. These new observations highlighted the importance of research in pesticide degradation by cold plasma. While the simple concept of pesticide degradation due to its exposure to plasma-generated reactive species is well-explained in the literature, research is needed in this area for an understanding of the key parameters and their effects on the decontamination process. Taking the present study as an example, the type of the plasma system can be considered as a key parameter. While DBD plasma systems have a general concept in producing plasma, the detail of their design, which varies depending on the supplier, can greatly affect its applicability for food processing (e.g., decontamination) (Gavahian et al., 2020). The same holds good for the arc plasma systems. Also, in the case of the arc systems, it should be noted that high temperatures of plasma gas may result in thermal degradation of pesticides, which may be wrongly interpreted as reactive species-induced degradation in some studies (Cengiz et al., 2015; Gavahian et al., 2019b). Besides, process parameters, including carrier gas composition, pressure, applied voltage, discharge distance, sample exposure mode (direct vs. indirect), can affect the performance of a plasma system for pesticide degradation.

Figure 3. Liquid chromatography-mass spectrometry (LC-MS) chromatogram of control (A), arc plasma-treated (B), and dielectric barrier discharge (DBD) plasma-treated (C) samples.

Zhou et al. (2018) reported that the performance of plasma in removing pesticide residues from food materials was related to the type of plasma and the process conditions such as the processing time and the plasma voltage. Likewise, Sarangapani et al. (2016) assessed the efficacy of high voltage DBD plasma on three types of pesticides, namely, endosulfan, malathion, and dichlorvos. According to the authors, the efficiency of high-voltage plasma in pesticide degradation varied, depending on the type of pesticide. They reported that 2 min of 60 kV plasma treatment resulted in 8.6, 7.9, and 4.2% decrease in the concentration of endosulfan, dichlorvos, and malathion, respectively. Therefore, different resistances to plasma conditions can be expected from different types of pesticides. Sarangapani et al. (2016) also showed that increasing both the input voltage and the treatment time resulted in better pesticide removal. For example, the input voltage of 80 kV and the extended process time eliminated up to 80.0% of dichlorvos from the pesticide sample. On the other hand, they observed that even applying high voltage for an extended time could not produce pesticide-free products, which suggests that cold plasma was not a reliable technique for the production of pesticide-free products, which is in line with the findings of the present study. Moreover, it was previously explained that different concentrations of reactive species can be produced by different types of plasma and also under different plasma conditions (Gavahian et al., 2018; Sarangapani et al., 2018). Furthermore, nonoptimized cold plasma treatment was shown to be inefficient in removing unpleasant allergens from milk (Tammineedi et al., 2013). Therefore, the efficacy of cold plasma in the degradation of pesticide residue should be fully explored before the industrial adaptation of this emerging technique, especially regarding plasma type, process condition, and the sensitivity of the target pesticide to the plasma treatment. A comparison between the effects of various plasma processes on chlorpyrifos is presented in Table 2. The type of plasma equipment and the process conditions are among the major effective parameters that should be considered in the plasma degradation of chlorpyrifos. It should also be noted that it should be the case for other technologies, such as Ultraviolet (UV), in spite of the promising results reported in the literature (Vasseghian et al., 2020).

Table 2. An overview of the effects of various plasma technologies on chlorpyrifos.

Plasma device Matrix Process condition Maximum degradation (%) Insight findings Reference
Monopole dielectric barrier discharge Apples – Plasma voltage: up to 13 kV
– Process time: up to 10 min
87 Plasma treatment significantly decreased pesticide concentration Mousavi et al., 2017
Gliding discharge plasma Mango – Argon was the carrier gas (flow rate: 5 L/min) 63 Gliding discharge plasma was introduced as an effective surface decontamination for mango Phan et al., 2018
Dielectric barrier discharge Tomatoes – Plasma power: up to 5 W
– Process time: up to 6 min
89 Dielectric barrier discharge plasma was introduced as an effective surface decontamination for tomatoes Ranjitha Gracy et al., 2019
Dielectric barrier discharge Maze – Argon was the carrier gas
– Plasma power: up to 25 W
– Process time: up to 1 min
91 Argon dielectric barrier discharge treatment was introduced to be effective in enhancing the corn safety Feng et al., 2019

Variations in sample temperature

Figure 4 represents fluctuations in the temperature of pesticide during 8 min of DBD and arc plasma treatments. While the temperature of samples remained steady in the DBD plasma system (due to the ability of this system in controlling the temperature), 8 min of arc plasma process altered the temperature of the pesticide sample from 30 to about 60°C at a rate of about 3.75°C.min–1. Likewise, it was observed that 10-min of plasma processing of water increased the temperature of water from 23 to 37°C (Ma et al., 2016). It should be noted that the temperature increase observed in the current investigation was also inefficient in pesticide degradation. It was previously explained that pesticides can stand mild temperatures (Amirahmadi et al., 2017; Kennedy and Mackie, 2018), which concurs with the observations in the present study.

Figure 4. Variations in the temperature of chlorpyrifos during arc and dielectric barrier discharge (DBD) plasma treatments.

Physical constants

The color parameters of the samples are presented in Table 3. According to the data, the plasma treatment did not significantly alter the lightness, blue–yellow, and green–red values of the sample. Researchers classified the overall color change (ΔE) value as great, well-visible, noticeable, slightly noticeable, and not noticeable when it ranged between 6.0–12.0, 3.0–6.0, 1.5–3.0, 0.5–1.5, and 0–0.5, respectively (Cserhalmi et al., 2006). It was previously explained that ΔE values above 3.0 are obvious to the human eyes (Puligundla et al., 2018). The data in Table 3 suggested that the variations in the overall color of the samples were not significant, considering the standard deviations of the reported values. Arc and DBD plasma treatments resulted in overall color change values of 1.1 ± 0.4 and 1.6 ± 1.2, respectively. Similar to the observations in this study, Dasan et al. (2018) reported that plasma treatment did not significantly affect the color of the eggshell samples (Dasan et al., 2018). Similar results were reported by Puligundla et al. (2018) for plasma-treated Gwamegi samples (Puligundla et al., 2018). Furthermore, Phan et al. (2018) disclosed that 10 min of arc plasma did not alter the color of a pesticide-contaminated sample, and all the overall color change (ΔE) was about 2.5. Likewise, the plasma process did not alter the refractive index of the pesticide solution, and all the samples had a similar refractive index of 1.3642. This finding was in line with the result of the LC-MS study. Similar physical properties of samples can be expected as arc and DBD plasma treatments did not alter the chemical structure of the samples. It should be noted that the decontamination efficiency and effects of plasma processing on the quality parameters of food samples should be assessed when an appropriate plasma system is designed and developed for degrading agrochemical residues of foods. These are among the key considerations for developing an effective plasma system that can be implemented in the industry.

Table 3. The effects of plasma treatments on the color parameters and refractive indexes of pesticides.

Control sample DBD plasma Arc plasma
RI* 1.3642a ± 0.0001 1.3642a ± 0.0001 1.3642a ± 0.0001
L* 20.64a ± 0.99 19.21a ± 0.41 20.29a ± 0.13
a* –0.36a ± 0.03 –0.39a ± 0.05 –0.37a ± 0.02
b* –2.36a ± 0.12 –2.21a ± 0.05 –2.33a ± 0.05
ΔE 1.56a ± 1.21 1.09a ± 0.35

*RI: Refractive index; L*: lightness; a*: greenness-redness; b*: blueness-yellowness; ΔE: overall color differences.

**Data are presented as Mean ± Standard deviation.

DBD, dielectric barrier discharge.


The effects of the two types of plasma devices, namely, DBD and arc, on a major pesticide residue of agricultural products, that is, chlorpyrifos, were investigated. LC-MS results revealed that the chlorpyrifos pesticide, under the experimental conditions used in this study, had high resistance against plasma treatments, i.e., neither arc nor DBD plasma treatments degraded this hazardous chemical. In spite of the promising decontamination effects of plasma reported in the literature (Table 2), the findings of this study can be considered prior to industrial adaptation of cold plasma for the production of pesticide-free products. Therefore, the commercial application of cold plasma for the removal of pesticides requires critical research and careful evaluation. Other aspects, such as selecting the appropriate type of plasma equipment, optimization of the process parameter, understanding the resistance of different types of pesticides against cold plasma, and further upscaling studies, should be considered. The use of plasma treatment to remove hazardous chemicals (e.g., pesticide pollutions) from products (e.g., fruits) with a waxy shell has not been well investigated in the literature. Therefore, the efficiency of plasma treatments in degrading other types of pesticides in such products can be considered further in prospective studies. Besides, the type of pesticide degrades are highly dependent on several parameters, such as plasma generation device, plasma power, plasma exposure time, and the carrier gas composition which influence the type and the concentration of the reactive species (e.g., reactive oxygen species) and the overall efficiency of plasma-induced degradation for a particular pesticide. Hence, the effects of other carrier gases, particularly O3, in plasma treatment should be examined to have a better understanding of the ability of cold plasma in removing residual toxins.


The authors would like to thank the College of Agriculture of National Pingtung University of Science and Technology (NPUST) of Taiwan for supporting plasma research. Also, the support of Food Industry Research and Development Institute of Taiwan is appreciated.


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