Materials Express

2158-5849/2019/9/451/008 Copyright © 2019 by American Scientific Publishers All rights reserved. doi:10.1166/mex.2019.1510 Printed in the United States of America www.aspbs.com/mex

Characteristics of prepared nano-emulsion and research on glutathione S-transferase enzymatic activity and penetration performance on T. cinnabarinus

Ni Yang1,†,TaoTang2, †,HuanYu1,FeiXue1, Chuanzhen Li1, 3, Shuang Rong1,LinHe1, 3,andKunQian1,∗ 1Key Laboratory of Entomology and Engineering, College of Plant Protection, Southwest University, Chongqing 400716, China 2State Key Laboratory for Quality and Safety of Agro-Products (In Prepared), Key Laboratory for Residue Detection of Ministry of , Institute of Quality and Standard for Agro-Products, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, Zhejiang, China

3Academy of Agricultural Sciences, Southwest University, Chongqing 400716, China Article

IP: 192.168.39.211 On: Sun, 26 Sep 2021 00:44:07 Copyright: American Scientific Publishers Delivered by Ingenta ABSTRACT In this study, fenpropathrin nano-emulsion (NE) was prepared by phase inversion emulsification method. 8% xylene and 2% solvent oil 150 (The main component is 1,2,4,5-Tetramethylbenzene) were used as solvent, styrylphenolpolyoxyethylene and calcium dodecylbenzenesulfonate were used as surfactants. The particle size, zeta potential, conductivity and contact angle were detected to evaluate the characteristic of the nano-emulsion. Toxicity of fenpropathrin nano-emulsion on the pest mite, Tetranychus cinnabarinus, was analyzed. The particle sizes of 8% fenpropathrin nano-emulsion and 20% fenpropathrin emulsion concentrates (EC) were 31.53 nm and 459.00 nm, zeta potentials were −22 mV and 5.762 mV, respectively, which showed that the size of nano- emulsion was much smaller and its stability was higher than that of EC. The contact angles of these two formulations were tested in concentrations of 500 mg/L to 2000 mg/L. We found that the contact angle of NE at the same concentration was 32% lower than that of EC averagely. The results indicated that the wettability and adhesion ability of nano-emulsion droplets were better than those of EC on the biological targets. According to determination of penetration performance to T. cinnabarinus, it was found that the penetration performance of NE to T. cinnabarinus is 4–6 times higher than that of EC. With the characteristics above, the NE has exhibited higher biological activity on the T. cinnabarinus. The results of Glutathione-S-transferase (GSTs) enzymatic activity of T. cinnabarinus showed nano-emulsion had higher effects than EC. In conclusion, compared with EC, nano-emulsion has better penetration, biological activity and a great application prospect in pesticide field in the future. Keywords: Fenpropathrin, Nano-Emulsion, Contact Angle, Enzymatic Activity, Penetration Performance.

1. INTRODUCTION Fenpropathrin acts on the nervous system of pests and is

∗ often used as / [1, 2]. Since its low Author to whom correspondence should be addressed.  Email: [email protected] solubility in water (14.1 g/L, 25 C), and unstability in †These two authors contributed equally to this work. alkaline solution, fenpropathrin is often used in the form

Mater. Express, Vol. 9, No. 5, 2019 451 Materials Express Characteristics of prepared fenpropathrin nano-emulsion Yang et al.

of emulsion concentrate (EC) for pest control. Unfortu- effects of fenpropathrin nano-emulsion and EC on GST nately, although EC is a traditional , activity of T. cinnabarinus. In addition, the differences of it has great negative effect on human health and the envi- contact angle, adhesion work and penetration performance ronment [3–5] because of the large quantities of organic between nano-emulsion and EC were also compared. solvents used in EC. In order to reduce environmental pol- Nano-emulsion is mainly used in cosmetics, medicine lution, it is necessary to replace EC with new environmen- and other fields, but it has not been promoted in pesti- tal protection formula. cide production. This experiment introduced the character- In recent years, nano- have become a research istics and advantages of nano-emulsion from many aspects hotspot. Many types of nano-pesticides are prepared by of pesticide properties, which is of great significance to using nano-materials as carriers [6]. Some researchers the application and popularization of nano-emulsion in the have developed new methods for pesticide detection by field of pesticide. using the adsorption properties of nano-materials, and improved the detection limit of pesticides [7–10]. Nano- 2. MATERIALS AND METHODS pesticides are the trend of pesticide development, and 2.1. Materials nano-emulsions are expected to become the hot dosage forms in the future. Nano-emulsions have a series of 8 wt.% fenpropathrin nano-emulsion was prepared by advantages such as small particle size, narrow droplet size a phase inversion emulsification method. Formula of distribution and so on, which make them very attractive in fenpropathrin nano-emulsion was as follows, 8% fen- many aspects of industrial applications [11–15]. Because propathrin, 8% xylene, 2% solvent oil 150, 11% emulsifier \ of the small particle size, nano-emulsion has high stability (LAE CaDDBS, 2:1) and 71% water. for precipitation, emulsification, flocculation or coagula- Fenpropathrin (emulsion concentrate, 20%) was pur- tion [16]. Although the particle size and some properties chased from Zhejiang Well-done Chemical Co., Ltd. of nano-emulsions are similar to those of microemulsions (Zhejiang, China). Fenpropathrin (technical grade, 92.9%) (for example, both being transparent) [17], the formation was provided by Nanjing Red Sun Co., Ltd. (Jiangsu, of nano-emulsions requires lower surfactant concentration China). Dimethylbenzene was purchased from Chengdu and less polar solvents [18–20]. Kelong Chemical Co., Ltd. (Chengdu, China). Calcium The key factors in the formationIP: 192.168.39.211 of nanoemulsion On: are Sun,dodecyl 26 Sep benzene2021 00:44:07 sulfonate (99%, CaDDBS) and poly- related to emulsification kinetics [21].Copyright: Nano emulsion American can Scientificoxyethylene Publishers fatty acid (99%, LAE) were provided by Delivered by Ingenta not spontaneously form. Their properties depend not only Lvshun Chemicals Co., Ltd. (Lvshun, China). All products on thermodynamic conditions, but also on the addition were not purified further. Deionized water was used in this experiment. Article method and order of components [22]. Stable and transpar- ent nano-emulsion can be prepared only by selecting suit- 1-chloro-2,4-dini-trobenzene (CDNB) was purchased able system composition and preparation method. In these from the Shanghai No. 1 Reagent Factory (Shanghai, studies, a low-energy method [23] was adopted at constant China); Coomassie blue G-250 was provided by Amresco temperature to prepare fenpropathrin (active ingredient) Co. (Solon, USA); bovine serum albumin (BSA) was from nano-emulsion with anionic-nonionic surfactant mixture. Shanghai BioLife Science & Technology Co. (Shanghai, The purpose of this work is to determine the optimum China). concentration and ratio of O/W nano-emulsion surfactant. The method is to test the influence of different propor- 2.2. Preparation Process of Nano-Emulsion tions and total concentration of surfactants on the long- In the preparation process of nano-emulsion, LEEM was term stability of nano-emulsion [24, 25]. Most researches used to prepare 20 g 8 wt.% fenpropathrin nano-emulsions are focused on the application of nano-emulsions in the with 8–12 wt% surfactant (emulsifier ABSA and phe-  medical fields [26], while very few are in the pesticide noxyethyl phenol polyoxyethylene ether) at 25 C. Firstly, field. fenpropathrin was completely dissolved in xylene, and In this study, a stable fenpropathrin nano-emulsion then surfactant was added into beaker to form oil phase was prepared by low-energy emulsification method evenly. And then water was added dropwise to oil phase (LEEM) [11, 27, 28] and the particle size, conductivity using a magnetic stirrer (JintanFuhua Instrument Co., Ltd., and zeta potential of the nano-emulsion were determined Jiangsu, China) for 30 min at 25 C. The preparation pro- for preliminary understanding the stability mechanism of cess of nano-emulsion by LEEM was shown in Figure 1. nano-emulsion. It can be predicted that pesticide micro- The stability of fenpropathrin nano-emulsion at 0 Cand droplets have better wettability and permeability to biolog- 54 C was analyzed. The experimental results were showed ical targets at nano-size, thus improving biological activity. in Table I. GSTs is one of the most important phase II metabolic enzymes in vivo, and is the main detoxification system of 2.3. Conductivity Measurement cell anti-injury. To a certain extent, it can reflect the bio- Conductivity was detected by a Conductimeter Crison logical activity of pesticides [29]. Therefore, we tested the model 525 with a Pt/platinized electrode.

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of Beibei District, Chongqing, China, and transferred to fresh potted young cowpea plants. The 200th generation after indoor breeding of about 13 years under artificial cli- mate: 24∼26 C, 60∼80% RH) was transferred into the centrifuge tube (three replicates for each treatment. After 24 hours, mites were observed under an anatomical micro- scope. If mites show immobility or irregular shaking of legs, they are considered dead.

2.7. Determination of Protein Concentration of Enzyme Source and GSTs Activity Fig. 1. Preparation process of fenpropathrin nano-emulsion by low- energy emulsification methods. Scheme of nano-emulsion preparation. Briefly, fenpropathrin (EC and NE) was dissolved in ace- −1 First, the original drug, solvents and surfactants are mixed evenly to tone (280 mg·L . Two hundred living female adult mites form an oil phase. Then water is dripped into the oil phase at room (3 to 5 days old) were chosen to conduct a determination temperature, and the nano-emulsion is formed by fully stirring. of GSTs activity in 6, 12 and 24 hours, respectively (three replicates for each sample with acetone as control). 2.4. Droplet Size and Zeta Potential Measurement With BSA as the standard [32], the total protein content Dynamic light scattering (DLS) was used to determine in enzyme solution was determined by Bradford method. Poly-dispersity index (PDI) and droplet size of the emul- The standard curve was firstly set using BSA. The 50 L of enzyme solution was mixed with Coomassie blue, sions by a Zetasizer Nano-ZS90 (Malvern Instruments −  · 1 ZEN3600, Malvern, UK) at 25 C employing an argon 0.1 mol L phosphatic buffer solution (PBS, pH 7.0) was = used as the control, and OD value was measured at 595 nm laser ( 633 nm). The nano-emulsion was diluted by  100 times with deionized water. after incubation at 37 C for 10 min.

Glutathione-S-transferase (GSTs) activity was deter- Article 2.5. Contact Angle Measurement mined according to the method modified by Hill, et al. [33, 34]. Two hundred female adult mites were Clean glass slide was placed onIP: Contact 192.168.39.211 angle measur- On: Sun, 26 Sep 2021 00:44:07 homogenized in 1.5 mL of PBS (0.04 mol·L−1,pH6.5)on ing instrument, and then 15 L of pesticideCopyright: solution American was Scientific Publishers ice. The supernatant was tested on ice after centrifugation dropped on the slide and the contact angles photosDelivered were by Ingenta at 10,000 r for 10 min at 4 C. CDNB (0.6 mmol·L−1 and taken with a CCD. Young Equation was used to calculate glutathione (GSH) (6 mol · L−1 were used as substrates, Contact angles (4 repeats for each treatment). The Young and GSTs reacted with reduced GSH after incubation for Equation is = + ×cos . refers to the free SV SL LV e SV 20 min at 37 C. The results were immediately recorded at energy of solid and vapour interface, refers to the SL intervals of 30 s for 5 min at 340 nm by microplate reader free energy of solid and liquid interface, , refers to the LV (three repeats for each sample). free energy of liquid and vapour interface and e means the contact angles. 2.8. Determination of Penetration Performance on T. cinnabarinus By Fenpropathrin 2.6. Bioassay of Fenpropathrin Nano-Emulsions on T. cinnabarinus Nano-Emulsion and EC 2.8.1. Fenpropathrin Treatment The modified residual coated vial (RCV) method was used to measure Median lethal concentration (LC50) The fenpropathrin (EC and NE) was dissolved in water · −1 values [30–31]. In this experiment, fenpropathrin nano- and diluted to 100 and 400 mg L . 2 mL was topically emulsion and emulsion concentrate were dissolved in applied to the experimental preparations. Four hundred liv- 100–1000 mg/L acetone to keep the mortality rate between ing female adult mites (3 to 5 days old) were chosen 20–80%. T. cinnabarinus (Three–five days old healthy to conduct a determination of fenpropathrin in 24 hours adult females T. cinnabarinus were collected in the field respectively (three replicates for each sample with water as control).

Table I. Stability of 8% fenpropathrin nano-emulsion after heat storage. 2.8.2. Sample Preparation Content of Content of It is difficult to collect enough hemolymph for amount of fenpropathrin before fenpropathrin after Decomposition fenpropathrin residue from adult T. cinnabarinus because Group heat storage (%) heat storage (%) rate (%) of the small body size. Therefore, we need to use the 1 7.823 7.632 2.4415 whole mite to extract and determine fenpropathrin. The 2 7.811 7.549 3.3542 living mites were collected and counted as one sample, 3 7.831 7.631 2.5540 Average 7.822 7.604 2.7870 all sample were washed 3 times with a colander in ace- tone, and were smashed thoroughly into mortar in liquid

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nitrogen. The sample was collected into a 10 mL cen- 3. RESULTS AND DISCUSSION trifuge tube after the addition of 2 mL acetone in a mortar. 3.1. Optimization of Nano-Emulsion The samples were sonicated for 5 min on the ice with Formulation Composition an ultrasonic processor (FS-450, 450 W, 20 KHZ, Shang When water was slowly added into the oil phase, the con- Chao Instruments PR China). The solution was centrifuged ductivity gradually changed. When 20% (4 mL) water  at 4 C for 10 min (10,000r), and then the clear super- was added, the conductivity quickly changed, which was natant extract was transferred into a 10 mL centrifuge tube. ascribed to phase transition from W/O nano-emulsion or Ultrasound-assisted extraction was repeated twice again liquid crystalline phase to O/W nano-emulsion (Fig. 2). with 2 mL acetone, acetone solution was introduced into The investigation of quality indexes showed that the same 10 mL centrifuge tube. The combined extracts the appearance of 8% fenpropathrin nano-emulsion was were evaporated to near dryness using a rotary vacuum evaporator at 25 C and reconstituted in 0.5 mL of ace- tone. Samples need to be filtered through a 0.22 m nylon syringe filter and transferred to an automatic injector vial for GC-MS injection.

2.8.3. GC–MS Analysis Equipped with a flame ionization detector (FID) and a 30 m fused film silica capillary column (the stationary phase is (5%-phenyl) methyl polysiloxane, 0.25 mm i.d., 0.25 mm film thickness), the GC–MS was performed on a Agilent 7890N gas chromatography linked to a 5977B mass selective detector (using 70 eV elective ionization). At 100 C, the oven temperature was hold for 1 min, and then raised to 240 C at a rate of 20 C/min, and then raised to 260 Catarateof2C/min, and hold for 12 min. IP: 192.168.39.211 On: Sun, 26 Sep 2021 00:44:07 The injection port was set at 260 C.Copyright: Fenpropathrin American was Scientific Publishers monitored at m/z 97, 125, 141 and 181. m/z 97 isDelivered selected by Ingenta as a quantitative ion. Fenpropathrin was quantified using calibration curves established using known quantities of Article standards. The concentrations of calibration standard are 0.01, 0.1, 0.25, 0.5, 1, the content of fenpropathrin was expressed as ng per individual.

Fig. 3. Size distribution and Zeta potential of 8% fenpropathrin NE and 20% fenpropathrin EC. (A) Size distribution of fenpropathrin NE (2000 mg/L); (B) Size distribution of fenpropathrin EC (2000 mg/L); (C) Zeta potential of fenpropathrin NE (2000 mg/L); (D) Zeta potential of fenpropathrin EC (2000 mg/L). (A and B) are particle size distri- butions of nano-emulsions and emulsifiable concentrates, respectively. Fig. 2. The effect of water amounts on conductivity of nano-emulsion. When the nano-emulsion and the emulsifiable oil are diluted with water, The curve represents the change of conductivity of nano-emulsion with small droplets will be formed and dispersed in the water. The particle the addition of water phase. First, the electrodes of the conductome- size distribution of small droplets will affect the stability of emulsion. ter are immersed in the oil phase, then the water phase is added at a (C and D) are Zeta potential maps of nano-emulsion and emulsifiable uniform speed and the conductivity values are recorded in time. When concentrate, respectively. Because of surfactant, Zeta potential can be 4 mL water was added, the conductivity quickly changed, which was generated on the surface of small droplets. The significance of Zeta ascribed to phase transition from W/O nano-emulsion or liquid crys- potential is that its value is related to the stability of the dispersion talline phase to O/W nano-emulsion. system.

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Fig. 4. Picture of contact angle of fenpropathrin nano-emulsions (NE) and emulsion concentrate (EC) (Different concentrations). This picture is the contact angles of nano-emulsion and emulsifiable concentrate on slide at different concentrations. The emulsions with different concentrations are added to the glass slide, and the angle between droplet and glass is contact angle after it is stabilized. The contact angle of a liquid on the surface of a solid material is an important parameter to measure the wettability of the liquid on the surface of the material. orange yellow transparent uniform emulsion, and it was narrow range, which was conducive to the stability of the stable under room temperature (25 C) storage for 14 d, emulsion. and can disperse rapidly in the course of 342 mg/L stan- Contact angle of emulsion samples was measured with dard hard water. The formulation is stable in cold and heat automatic contact Angle meter. The results were shown in storage (Table I). Figure 4. Compared with EC, the Contact Angle of nano- emulsion was much smaller, which showed that the nano- 3.2. Droplet Size and Zeta Potential of emulsion has ultra-low oil/water interfacial tension value, Article Fenpropathrin Nano-Emulsions and can effectively reduce the contact angle of the droplets The particle size distributionIP: of 192.168.39.211 two kinds of emul- On: Sun,on 26 the Sep plants. 2021 Adhesion 00:44:07 work refers to the degree of adhe- Copyright: American Scientificsion of two Publishers phases contacting each other on the interface sion samples was determined by laser particle sizeDelivered tester by Ingenta (diluted to 2000 mg/L). Average particle size of nano- layer. Test of wetting adhesion has been done to determine emulsion was 31.53 nm, and the polydispersivity index if spray droplets can effectively adhere to the plant leaves. (PDI) was 0.123, while the average particle size of fen- Adhesion work was maximum work in the wet procedure. propathrin EC was 459.0 nm, and the PDI is 0.658 The greater the adhesion work was in the interface of solid (Figs. 3(A and B)). Compared with EC, droplet size dis- and liquid, the more firmly it was. Table II showed that tribution of nano-emulsion was in a narrow range, which the adhesion work of nano-emulsion was obviously higher was conducive to stability of the emulsion. than that of EC, which indicated that the nano-emulsion Zeta potential of emulsion samples was determined by droplets can adhere to the targets more firmly, and more laser particle size tester (diluted to 2000 mg/L), and the pesticide can be absorbed into the targets and the control result was shown in Figure 3(D). The Zeta potential value efficacy can be improved. of nano-emulsion was −22 mV, and the negative charges of the colloidal particles surface were mainly ascribed 3.3. Biological Activity and GST Enzymatic Activity to anionic emulsifier CaDDBS. The absolute Zeta poten- Assay of Fenpropathrin Nano-Emulsions on tial value of fenpropathrin nano-emulsion was obviously T. cinnabarinus higher than EC. The higher potential value can effectively The toxicities of fenpropathrin with different formulations prevent the cohesion between particles. Meanwhile, Elec- to Tetranychus cinnabarinus were showed in Table III. trostatic repulsion can also keep the system in good and The results showed that LC50 of NE to Tetranychus stable station. From Figures 3(C) and (D), it can be seen cinnabarinus was 484.87 mg/L, while LC50 of EC was that the Zeta potential of nano-emulsion distributed in a

Table III. The toxicity of fenpropathrin with different formulations to Table II. Adhesion work of different concentrations droplets on glass. Tetranychus cinnabarinus (Boisduval).

Fenpropathrin Adhesin work Fenpropathrin Adhesin work Toxicity × −3 2 × −3 2 NE (mg/L) ( 10 J/m )EC(mg/L)(10 J/m ) Fenpropathrin regression LC50 95% confidence Correlative formulations equation (mg/L) limit (mg/L) coefficient 500 114.430 ± 8.325 500 92.676 ± 8.919 1000 118.527 ± 11.304 1000 104.538 ± 3.624 NE Y = 127X + 159 484.87 382.11∼615.27 0.9815 2000 130.932 ± 5.179 2000 106.168 ± 5.657 EC Y = 289X − 311 641.09 569.24∼772.01 0.9533

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1.2 time, small liquid droplets can reduce the rebound of Ck Fenpropathrin NE Fenpropathrin EC the droplets in the foliage and drift of effective ingredi- 1 b b b c ents [35]. b b 0.8 The effects of fenpropathrin nano-emulsion and EC on

mol/mg/min) GST enzymatic activity of T. cinnabarinus at 6 h, 12 h, µ a 0.6 24 h was showed in Figure 5. At 6 h, there was no sig- a a nificant difference between the relative activities of GST 0.4 treated with fenpropathrin NE and EC, while they were both higher than that of the control. At 12 h, the relative 0.2 activity of GST treated with fenpropathrin NE was higher Relative activity ( 0 than those of CK and EC, which showed fenpropathrin 6h 12h 24h NE had stronger effects on T. cinnabarinus to increase the Time (h) GST enzymatic activity. While at 24 h, the relative activ- ities of GST treated with fenpropathrin NE and EC were Fig. 5. The effects of fenpropathrin nano-emulsion and EC on GST enzymatic activity of T. cinnabarinus at 6 h, 12 h and 24 h. The not significantly different, which showed GST enzymatic effects of nano-emulsion and emulsifiable concentrate on GST activity activity recovered gradually after 24 h of treatment with of Tetranychus cinnabar at different times are shown in this picture. fenpropathrin NE and EC. T. cinnabarinus was treated by membrane method and the GST activity was measured at 6, 12 and 24 hours. 3.4. The Effects of Fenpropathrin Nano-Emulsion and EC on Penetration Performance on 552.66 mg/L. The toxicity of the NE to Tetranychus T. cinnabarinus cinnabarinus was much higher than that of the EC. Due to First, the GC/MS-SCAN model was used to analyze fen- nanolevel droplets, ultra-low interfacial tension and small propathrin standard. The total ionic current (TIC) (A) and contact angle of nano-emulsion, the pesticide solution was mass spectrum (B) of fenpropathrin were obtained respec- easier to spread out on the leaves, and more solution tively. As shown in Figure 6(A), the retention time of fen- can penetrate into crop and target insects. At the same propathrin is 13.117 min. The unique fragment ion peaks IP: 192.168.39.211 On: Sun, 26 Sep 2021 00:44:07 Copyright: American Scientific Publishers Delivered by Ingenta Article

Fig. 6. The total ionic current (A) and mass spectrum (B) of fenpropathrin standard samples, and the total ionic current (C) and mass spectrum (D) of fenpropathrin samples. (A) is the total ionic current (TIC) of fenpropathrin standard and (B) is mass spectrum of fenpropathrin standard. Both (A and B) were obtained by GC-MS analysis of fenpropathrin standard. (C and D) are the total ionic current (TIC) and mass spectrum of fenpropathrin sample, respectively. The fenpropathrin (EC and NE) was dissolved in water and sprayed on the leaves which have T. cinnabarinus. After the liquid was dried, T. cinnabarinus was washed in acetone three times to remove fenpropathrin from the surface of T. cinnabarinus. Then, fenpropathrin samples were prepared by comminuting T. cinnabarinusin liquid nitrogen and extracting volume with acetone. Finally, the content of fenpropathrin in the samples was determined by GC-MS.

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Table IV. Recovery test for fenpropathrin. of NE. The residue of fenpropathrin was 0.2127 ± 0.0358 ng/individual with the same concentration treat- Amount of Detection addition (g) value (g) Recovery (%) RSD (%) ment of EC. Under 100 mg/L treatment, the residual amounts of NE and EC were 0.5421±0.1954 ng/individual 2.0 1 783 89 15 1 00 and 0.0878 ± 0.0765 ng/individual, respectively. There- 1.0 0862 8620 224 0.2 0171 8550 576 fore, the penetration performance of NE to Tetranychus cinnabarinus is better than that of EC with the same con- centration. For the fenpropathrin NE, it is easier to adhere (Fig. 6(B)) can be obtained: m/z = 97, 125, 141, 181 and and penetrate into target insects. 97 as base peak ions. Therefore, we chose m/z = 97 for quantitative ions and the rest for reference ions, and for 4. CONCLUSIONS selective ion monitoring, the scanning time was 17 min. In this research, fenpropathrin nano-emulsion was pre- The standard concentration liquid of fenpropathrin was pared in phase inversion emulsification. With higher sta- diluted in sequence to prepare the standard solution of bility, smaller contact angle and higher adhesion work, the 1, 0.5, 0.25, 0.1 and 0.01 mg/L, and the peak area was fenpropathrin nano-emulsion showed higher control effi- recorded by characteristic ion 97. Each concentration of cacy than EC did. Moreover, NE had better penetration standard liquid was injected 3 times. The regression equa- performance and the ability to influence GST enzymatic y = x + 2 = tion can be obtained: 614 17 14 976 r 0.982, activity of T. cinnabarinus than EC did. With the character- X Y ( , mg/L) is concentration, is the peak area) and the istics above, the NE has exhibited higher biological activ- linear range is 0.01–1 mg/L. ity on the T. cinnabarinus. With reduction of hydrophilic Then, the samples were analyzed by GC/MS-SCAN and hydrophobic solvent, the fenpropathrin nano-emulsion mode, and the total ion current (C) and mass spectrum (D) is friendlier to environment. The study on fenpropathrin were obtained respectively. By comparing Figures 6(B) nano-emulsion also can be used as reference to develop and (D), the results show that the samples have the same other pesticide nano-emulsion. Article unique fragment ion peaks as the standard ones. And as shown in Figures 6(A) and (C), there are obvious peaks in IP: 192.168.39.211 On: Sun,Acknowledgments: 26 Sep 2021 00:44:07This work was supported by the both the sample and the standard at 13 min, indicating the Copyright: American ScientificNational Publishers Key Research and Development Program of presence of fenpropathrin in the samples. Delivered by Ingenta China (No: 2017YFD0200301), the Fundamental Research The relative standard deviations (RSD) of the intraday Funds for the Central Universities (No: XDJK2017B022), sample and interday samples were obtained by sampling and the significant science and technology projects of 6 times on the same day and sampling once each day in Yunnan (2018ZG008). six days, respectively. The RSD of intraday and interday samples were 2.76% and 6.74%, respectively. Three concentrations of fenpropathrin standard solution References and Notes were prepared. And in accordance with the 2.8.2 method 1. Kanawi, E., Budd, R. and Tjeerdema, R.S., 2013. Environmental for processing extraction, the concentration of each sample fate and ecotoxicology of fenpropathrin. Reviews of Environmental solution was detected by GC-MS for three times, and the Contamination and Toxicology, 225, pp.77–93. 2. Nooshin, Z.S. and Leila, R., 2015. Evaluation of five essential oils results were recorded in Table IV. The average recovery as botanical against the strawberry spider mite tetrany- rate is 86.95%. chus turkestani ugarov and nikolskii. International Biodeterioration The penetration performances of fenpropathrin with & Biodegradation, 98, pp.101–106. different formulations to Tetranychus cinnabarinus were 3. Durre, S., Ifthikhar, H., Maqsood, A., Mirza, A.B., Atufa, K. and showed in Table V. The results showed that the Faraz, A., 2015. Development of cereal baits and comparative field efficacy of some additives as bait carrier for zinc phosphide and residue of fenpropathrin in Tetranychus cinnabarinus was coumatetralyl against rodent pests of poultry farms in Rawalpindi– 0.8294 ± 0.2905 ng/individual with 400 mg/L treatment Islamabad, Pakistan. International Biodeterioration & Biodegrada- tion, 104, pp.460–471. 4. Liu, L., Zuo, M., Cheng, J., Matsadiq, G., Zhou, H.B. and Li, J.K., Table V. Fenpropathrin content in T. cinnabarinus of different concen- 2011. Coupling polymer monolith microextraction to gas chromatog- trations NE and EC. raphy: Determination of in water samples. Microchimica Acta, 173(1–2), pp.127–133. Sample The content of 5. Michael, W.F. and William, B.J., 2000. Future technology for man- concentration fenpropathrin T. cinnabarinus aging problems with vertebrate pests and over-abundant wildlife Sample type (mg/L) in (ng/individual) an introduction. International Biodeterioration & Biodegradation, High concentration NE 400 0.8294 ± 0.2905a 45(3–4), pp.93–95. Low concentration NE 100 0.5421 ± 0.1954ab 6. Kunanon, C., Chompoonut R. and Darinee P., 2018. Investigation High concentration EC 400 0.2127 ± 0.0358bc of functional graphene/ pesticide molecules by using Low concentration EC 100 0.0878 ± 0.0765c density functional theory calculation. Journal of Nanoscience and Nanotechnology, 18(10), pp.6786–6790.

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7. Yuan, C.X., Qian, Y.W., Hong, X., Wang, H., He, H.N., Liu, H.L., 22. Isabel S., Maestro, A., González, C., Solans, C. and Gutiérrez, Chai, Z.L., Zhang, Y.J., Zhao, P. and Wang, Y., 2019.Anew J.M., 2006. Optimization of nano-emulsion preparation by low- reversed-dispersive micro-solid-phase extraction of organophospho- energy methods in an ionic surfactant system. Langmuir, 22(20), rus pesticides based on three-dimensional magnetic nanoparticles pp.8326–8332. supported by graphene-carbon nanotubes nanocomposite. Journal of 23. Ee, S.L., Duan, X.M., Liew, J. and Nguyen, D.Q., 2008. Droplet Biobased Materials and Bioenergy, 13 (2), pp.170–174. size and stability of nano-emulsions produced by the temperature 8. Liu, K., Dong, H. and Deng, Y., 2016. Recent advances on rapid phase inversion method. Chemical Engineering Journal, 140(1–3), detection of pesticides based on enzyme biosensor of nanomaterials. pp.626–631. Journal of Nanoscience and Nanotechnology, 16(7), pp.6648–6656. 24. Fernandez, P., André, V., Rieger, J. and Kühnle, A., 2004. Nano- 9. Deng, Y., Liu, K., Liu, Y., Dong, H. and Li, S., 2016. An novel emulsion formation by emulsion phase inversion. Colloids & acetylcholinesterase biosensor based on nano-porous pseudo car- Surfaces A Physicochemical & Engineering Aspects, 251(1–3), bon paste electrode modified with gold nanoparticles for detection pp.53–58. of methyl . Journal of Nanoscience and Nanotechnology, 25. Bouchemal, K., Briançon, S., Perrier, E. and Fessi, H., 2004. Nano- 16(9), pp.9460–9467. emulsion formulation using spontaneous emulsification: Solvent oil 10. Liu, Y., Liu, K., Dong, H., Zhang, L., Deng, Y., Ma, C. and Wang, Z., and surfactant optimisation. International Journal of Pharmaceutics, 2016. Acetylcholinesterase biosensor for detection based on 280(1), pp.241–251. nano-porous pseudo carbon paste electrode. Nanoscience and Nan- 26. Pant, M., Dubey, S., Patanjali, P.K., Naik, S.N., Sharma, S. and otechnology Letters, 8(9), pp.785–790. Satyawati, S., 2014. Insecticidal activity of eucalyptus oil nanoemul- 11. Subramanian, B., Kuo, F., Ada, E., Kotyla, T., Wilson, T., sion with karanja and jatropha aqueous filtrates. International Biode- Yoganathan, S. and Nicolosi, R., 2008. Enhancement of anti- terioration & Biodegradation, 91, pp.119–127. inflammatory property of aspirin in mice by ananoemulsion prepa- 27. Ahmed, M., Smith, D.M., Hamouda, T., Rangelmoreno, J., ration. International Immunopharmacol, 8(11), pp.1533–1539. Fattom, A. and Khader, S.A., 2017. A novel nanoemulsion vac- 12. Anton, N., Gayet, P., Benoit, J.P. and Saulnier, P., 2007. Nano- cine induces mucosal interleukin-17 responses and confers protec- emulsions and nanocapsules by the PIT method: An investigation on tion upon mycobacterium tuberculosis challenge in mice. Vaccine, the role of the temperature cycling on the emulsion phase inversion. 35(37), pp.4983–4989. International Journal of Pharmaceutic, 344(1), pp.44–52. 28. Liu,W.R.,Sun,D.J.,Li,C.F.,Liu,Q.andXu,J.,2006.Forma- 13. Forgiarini, A., Esquena, J., Gonzlez, C. and Solans, C., 2011.For- tion and stability of paraffin oil-in-water nano-emulsions prepared by mation of nano-emulsions by low-energy emulsification methods at the emulsion inversion point method. Journal of Colloid Interface, constant temperature. Langmuir, 17(7), pp.2076–2083. 303(2), pp.557–563. 14. Gutiérrez, J.M., González, C., Maestro, A., Solè, I., Pey, C.M. and 29. Pey, C.M., Maestro, A., Solé, I., González, C., Solans, C. and Nolla, J., 2008. Nano-emulsions: New applications and optimization Gutiérrez, J.M., 2006. Optimization of nano-emulsions prepared by of their preparation. Current OpinionIP: in Colloid192.168.39.211 & Interface Science, On: Sun, 26low-energy Sep 2021 emulsification 00:44:07 methods at constant temperature using a 13(4), pp.245–251. Copyright: American Scientificfactorial Publishers design study. Colloids & Surfaces a Physicochemical & 15. Solans, C., Izquierdo, P., Nolla, J., Azemar, N. and Garcia-Celma,Delivered by IngentaEngineering Aspects, 288(1–3), pp.144–150. M.J., 2005. Nano-emulsions. Current Opinion in Colloid & Interface 30. Sheehan, D., Meade, G., Foley, V.M. and Dowd, C.A., 2001.Struc- Science, 10(3–4), pp.102–110. ture, function and evolution of glutathione transferases: Implications

Article 16. Tadros, T., Izquierdo, P., Esquena, J. and Solans, C., 2004. Formation for classification of non-mammalian members of an ancientenzyme and stability of nano-emulsions. Advances in Colloid and Interface superfamily. Biochemical Journal, 360(1), pp.1–16. Science, 108, pp.303–318. 31. Leeuwen, V.T., Pottelberge, V.S., Nauen, R. and Tirry, L., 2007. 17. Katagi, S., Kimura, Y. and Adachi, S., 2007. Continuous preparation and acaricides antagonise bifenazate of O/W nano-emulsion by the treatment of a coarse emulsion under toxicity through esterase inhibition in tetranychusurticae. Pest Man- subcritical water conditions. LWT-Food Science and Technology, agement Science, 63(12), pp.1172–1177. 40(8), pp.1376–1380. 32. Feng, Y.N., Zhao, S., Sun, W., Li, M., Lu, W.C. and He, L., 2011. 18. Morales, D., Gutiérrez, J.M., Garcıa-Celma, M.J. and Solans, Y.C., The sodium channel gene in tetranychus cinnabarinus (boisduval): 2003. A study of the relation between bicontinuous microemul- Identification and expression analysis of a mutation associated with sions and oil/water nano-emulsion formation. Langmuir, 19(18), resistance. Pest Management Science, 67(8), pp.904–912. pp.7196–7200. 33. Bradford, M.M., 1976. A rapid and sensitive method for the quan- 19. Izquierdo, P., Esquena, J., Tadros, T.F., Dederen, J.C., Feng, J., titaion of microgram quantities of protein utilizing the princi- Garcia-Celma, M.J., Azemar, N. and Solans, C., 2004. Phase behav- ple of protein-dye binding. Analytical Biochemistry, 72(248–25), ior and nano-emulsion formation by the phase inversion temperature pp.248–254. method. Langmuir, 20(16), pp.6594–6598. 34. Habig, W.H., Pabst, M.J. and Jakoby, W.B., 1974. Glutathione 20. Sadurn, N., Solans, C., Azemar, N. and Maria JoseGarcˇ ˇia-Celma S-transferases, the first step in mercapturic acid formation. Journal 2005. Studies on the formation of O/W nano-emulsions by low- of Biological Chemistry, 249(22), pp.7130–7139. energy emulsification methods suitable for pharmaceutical applica- 35. Hill, C.B. and Hartman, G.L., 2004. Resistance to the soybean aphid tions. Acta Biochimica Polonica, 26(5), pp.438–445. in soybean germpl-asm. Crop Science, 44(1), pp.98–106. 21. Atanase, L.I., Larraya, C., Tranchant, J.F. and Save, M., 2017. 36. Shen, Y., Cai, H.Z. and Shen, X.B., 2009. Research on resistance of Rational design of tetrahydrogeraniol-based hydrophobically modi- german cockroach against DDVP nanoemulsion and DDVP ordinary fied poly(acrylic acid) as emulsifier of terpene-in-water transparent emulsion. Journal of Environmental & Occupational Medicine, 4(1), nanoemulsions. European Polymer Journal, 94, pp.248–258. pp.349–352.

Received: 1 June 2019. Accepted: 4 June 2019.

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