Authentication of Microplastic Accumulation in Customary Fruits and Vegetables


 The environment has become a major source of plastic pollution. Microplastics have been well documented in aquatic ecosystems as an increasing pollutant of worldwide relevance, but little is known about their effects on the terrestrial environment, particularly in agroecosystems. Researches have recently proven that microplastics can affect human health and are found in human organs and tissues. In this present study, two different types of fruits like grapes (Vitis vinifera) and banana (Musa paradisiaca), vegetables like brinjal (Solanum melongena) and potato (Solanum tuberosum) were collected from local market in Trichy, Tamil Nadu, India and analysed for microplastics accumulation. Further, we identified the microplastic size through stereomicroscope, in grapes and banana the microplastic size was 0.002 mm and 0.01 mm respectively as well as vegetables like potato and brinjal the microplastic size was 0.002 mm and 0.01mm. The identified microplastics had been chemically characterized by FT-IR and viewed through SEM.


Introduction
Plastic pollution has become a major scienti c and societal problem. Because of the rapid increase in plastic manufacture and consumption. Despite a gradual growth in plastic recycling and energy recovery, the majority of plastic wastes are released into the environment on a global scale (Blasing and Amelung, 2018). Microplastics (MPs) have recently been the subject of environmental research due to their small size and potential impact on terrestrial ecosystems. Microplastic is described as a plastic with a diameter of less than 5 mm and comes in a range of morphologies, including beads, fragments, bres, and lms. MPs are divided into two categories: primary MPs, which are formed in small quantities and discharged into the environment, and secondary MPs, which are degraded from bigger plastic wastes due to water, wind, sunshine, and other environmental factors. Primary MPs are released directly into the environment, whereas secondary MPs are formed by the breakdown of large-size chunks. Microplastics can enter agroecosystems through a variety of routes, including fertiliser coatings (Heuchan et al., 2019), wastewater irrigation (Zhang and Liu, 2018), compost addition and biosolids application (Nizzetto et al., 2016;Weithmann et al., 2018), and, most importantly, the use of mulching lm (Nizzetto et al., 2016). ( Agricultural systems are the nal bene ciaries of a variety of contaminants, including microplastics, whose impacts are relatively unclear (Razzaghi et al., 2018). In general, there is a lack of information of material fate and detrimental consequences in the agricultural system, which leads to food chain failure and an unknown channel of human exposure. Microplastics, depending on their size and type, can penetrate the seed, root, culm, leaves, and fruit cells (Dietz and Hertz, 2011). There is very few microplastic accumulation studies on regular consumed fruits and vegetables so we focused in this present study.
Currently, lacking of information on this subject due to the absence of data about the microplastics presence in edible fruits and vegetables. So we decided and applied our method to evaluate the microplastics <5mm presence in common edible fruits and vegetables. The aim of this study is to investigate the presence of microplastics present in regularly consumed vegetables like brinjal (Solanum tuberosum) and potato (Solanum melongena) and fruits of grapes (Vitis vinifera) and banana (Musa paradisiaca) and also identi ed and characterized the microplastics through various analytical methods.

Sample Collection and Preparation
Two types of samples like fruits (Grapes and Banana) and vegetables (Brinjal and Potato) were collected from local market at Tiruchirapalli, Tamil Nadu, India. Samples were put into cleaned aluminium tray and kept in closed conditions. Collected samples were sliced in the same day by using sterilized steel knife.
The sliced fruits and vegetables samples were kept on clean sterilized petri plates. Then the samples were dried in hot air oven, at 60 C temperature for one day. The dried samples were powdered using clean sterilized mortar and pestle. Then grained samples were air dried for four days after that samples were extracted. Only glass equipment and containers were utilised, any plastic material and any product with an inorganic carbon chemical structure (containers, caps, pipettes, lters, holders) were avoided. Still no standardized protocol for extracting microplastics in fruits and vegetables. Therefore, we implemented a methodology (Fabio Corradini et al., 2019).
The dried fruit and vegetable samples were weighed at 5g and deposited in glass centrifuge tubes with a capacity of 50 ml. Each tube was added 20 ml of deionized water, and the samples were swirled for 30 seconds. The supernatant was ltered using Whatmann No.42 lter paper after centrifugation at 2000 rpm for 15 minutes. The precipitate was agitated and centrifuged after 20 ml of sodium chloride was added. The supernatant was ltered through the same Whatmann No.42 lter paper for the second time.
For the nal extraction, 20 ml of a concentrated zinc chloride solution was added to the centrifuge tubes with the precipitate. The samples were swirled for 30 seconds since the ZnCl2 solution had a higher viscosity than the other solutions. The supernatant was ltered using Whatmann No.42 lter paper after centrifugation at 2000 rpm for 15 minutes. After that, lter paper were placed on Petri dish for optical inspection in stereo microscope (Fabio Corradini

FT-IR Analysis
Fourier transform infrared spectroscopy can provide a distinct infrared spectrum for a speci c chemical bond. Various samples have different bond compositions, so its is used to identify an unknown substance by comparing its spectrum with the spectra of known materials. FT-IR has become one of the most widely utilised techniques in chemical characterization of microplastics due to its great dependability. The visible microplastic particles in fruits and vegetables extracted samples were taken for FT-IR analysis (Perkin Elmer, Spectrum RX, USA). Spectra in the frequency range of 4000-400 cm −1 was used to record the spectrum of the extracted microplastics (Jung et al., 2018).

Scanning Electron Microscope Analysis
The extracted microplastic particles were taken by the previously de ned method (Fabio corradini et al., 2019). The samples were gold-coated on the surfaces of microplastic particles while micrographs were taken. After xation, the samples were placed in a deep vacuum and examined using a scanning electron microscope (SEM) (ZEISS EVO, Carl Zeiss microscopy). The SEM photograph had revealed that the microplastic present in the samples. The SEM micrograph also showed at high power of the electron beam.

Sample Collection and Preparation
This study reports were revealed the presence of microplastics in regular intake of edible fruits and vegetables. Fruits and vegetable samples were collected and dried to sieved for extraction of microplastics particles.

Stereomicroscopic Analysis
The microplastic accumulation in fruits and vegetables sample was identi ed through stereomicroscope.
In stereomicroscope microplastic size was measured, grape fruit microplastic size is 0.002 mm and banana 0.01 mm and vegetables samples like potato microplastic size is 0.002 mm and brinjal size is 0.01 mm ( Figure.1).

Characterization of Microplastics
Extracted microplastics were transferred to the FT-IR spectra to determine the different chemical groups were present in fruits and vegetables. In banana sample has shown spectral range at 3444.68 cm −1 , 1633.55 cm −1 refers to C=O stretching, 693 cm −1 peak range represent aromatic CH and 1271.00 cm −1 peak range refers to C-N stretching. In grapes sample has shown spectral range at 3436.06 cm −1 and 1633.82 cm −1 refers to C=O stretching, 1270.76 cm −1 refers to NH bending. In potato sample has shown spectral range at 3435.84 cm −1 and 1634.11 cm −1 represents C=O stretching,684 cm −1 represents C=O bending. In brinjal sample has shown spectral range at 3435.82 cm −1 C=O stretching,1120.93 cm −1 and 696 cm −1 peak range represent acrylonitrile butadiene styrene (ABS) group (Table.1 and Figure 2).

Scanning Electron Microscope Analysis
Electron images of microplastic particles (a, b, c, d) were shown in gure.3. Resulting spectra of the SEM analysis demonstrated the presence of microplastics presence in fruits and vegetables. In SEM results were show that microplastic size was found in fruits like grapes 2 µm and banana is 10µm.Vegetables like potato sample microplastic size is 2 µm and brinjal microplastic size is 10 µm.

Discussion
Microplastics have long been recognised as a serious environmental problem due to their widespread use and slow decomposition. In our experiment we detected the microplastics accumulation in fruits and vegetables. Two types of fruits and vegetables were taken and analyzed for microplastics accumulation.
It is the preliminary study for accumulation of microplastics.
In this study, we have detected the MPs at 2µm and 10 µm size in fruits and 2µm and 10 µm size in vegetables respectively. The size of the MPs is less than 5mm in diameter a similar study was reported by refers to C=O stretching resulted nylon polymer, spectral peak at 1250-1350 cm −1 shows C-N stretching resulted high density polyethylene terephthalate polymer group present on it. Similar peaks were found in our banana sample shows peak range 1271.00 cm −1 has C-N stretching it refers to high density polyethylene terephthalate polymer group. At 1050 cm −1 -1150 cm −1 peak range C-N stretch was found and refers to polyethylene oxide group. Same results were found in banana sample at range 1121.33 cm −1 shows C-N stretch it refers to polyethylene oxide group. At peak range 1050 cm −1 -1150 cm −1 C-N stretch was found and refers to polyethylene oxide group. In brinjal same results found in the peak range 1120.93 cm −1 and refer to polyethylene oxide group.

Conclusion
The present study was mainly focused on microplastic accumulation in edible fruits and vegetables. We investigate the presence of microplastics in vegetables (potato and brinjal) and fruits and to identi ed and characterized the microplastics group through various analytical methods like FTIR and SEM. This study recommends more studies to examine the risks of human exposure to microplastics due to ingestion of agricultural products. In addition, agricultural impacts of microplastics can be complicated by their ability to absorb other chemicals, and the events of climate change due to anthropogenic factors, which future studies could aim to investigate. Government organisations and health authorities must act quickly to enact and implement environmental rules that will oversee the manufacturing, use, and disposal of plastics.