Nanoparticle Technology of Dodecenol As Active Components in © 2019 Subekti N, Et Al

Research Article: Open Access Accepted: September12,2019 Subekti etal. Tel: +62-024-8508033 author: *Corresponding Introduction 2 1 Niken Subekti which is caused by caused is which [1, year per loses rupiah billion than are economically very harmful. Economically, more cynocephalus Rahmawati Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, in provided theoriginalauthorand sourcearecredited. reproduction and distribution, use, unrestricted permits which License, Attribution Commons Copyright: Chemistry Department,UniversitasNegeriSemarang,Indonesia Biology Department,UniversitasNegeriSemarang,Indonesia Cryptotermes Cynocephalus Citation: Components in In Indonesia drywood termite or Cryptotermes termite drywood Indonesia In Nanoparticle TechnologyofDodecenolasActive Subekti N, Mar’ah R, Aisyah AN, Nada DF, Rahmawati M (2019) Nanoparticle Technology of Dodecenol as Active Components in © 2019 Subekti N, et al. This is an open-access article distributed under the terms of the Creative the of terms the under distributed article open-access an is This al. et N, Subekti 2019 © IntJNanoparticlesNanotech2019, 5:031 VIBGYOR preference topinewoodsampleindicatedbymassloss13.856%. with a significance level of 5%. Petridis assay shows that nanoparticle 10% maximally increases termite SGE than influential more is BE termites. of behavior the on SGE with BE of effect the in difference a is SGE effect on termite adaptation and termite (P < 0.05). The bioassay choice test of Y shows that there and BE between difference a was there that showed test Y Bioassay compounds. dodecenol are there extract termite the in that show results chromatogram GC-MS The repetitions. 3 and 25%) 15%, and 10%, 20%, 5%, (0%, concentration nanoparticle dodecenol of treatment 6 used research this in test endurance and assay Petridis 2006. Standard National Indonesian on based test endurance and assay dish Petri using conducted was test food-preference The gel. nanoparticle produce to material main a as used then was BE termites. of behavior feeding on effect its compare to Y assay bios tested then GC-MS using analyzed were extracts Both both. of components chemical of analysis and preference Technology, Nanoparticle,Dodecenol,Drywood-termites Keywords controlling for (SGE) Extract Gland Sternal and (BE) Extract Body from dodecenol pheromone nanoparticle of effectiveness the analyze to aims study This pheromones. dodecenol of nanotechnology is controlling for used be can that technologies the of One year. per rupiah of billions to up losses economic incurred Damages pest. wood-destroying as Cryptotermes cynocephalus Abstract 2 has been recorded as termites that termites as recorded been has 1* , Ro’iyyatulMar’ah C. cynocephalus Niken Subekti, Biology Department, Universitas Negeri Semarang, Indonesia, Semarang, Negeri Universitas Department, Biology Subekti, Niken Control System. IntJNanoparticles Nanotech5:031 ; Published: Cryptotermes CynocephalusControl September14,2019 is one of the Kalotermitidae species. In Indonesia, this species is known is species this Indonesia, In species. Kalotermitidae the of one is lead ]. The losses The 2]. System people al- people 1 , AfrinNurAisyah Nanoparticles andNanotechnology International Journal of Cryptotermes cynocephalus cephalu able to kill approximately 55.67% dry wood is acid boric using preservation wood that showed results The termiticide. uses generally Indonesia ways lookfortocontrolthetermitegrowth[ The control of dry wood termites carried out in out carried termites wood dry of control The [4]. However, termiticides it is not environ- 1 , DinaFikaNada through termite feeding termite through 2 andMeidina ISSN: 2631-5084 C. cyno C. 3]. - Subekti et al. Int J Nanoparticles Nanotech 2019, 5:031 ISSN: 2631-5084 | • Page 2 of 6 • mentally friendly and likely extinguish non-target made from the whole body of the termites (Body organisms. Besides that, the termiticide product, Extract/BE) and the Stern Gland Extract (SGE). BE which is also widely used today is diflubenzuron was produced by extracting 2000 adult individuals and hexaflumuron formulated in bait form of. of pseudergate using 20 ml of hexane at 4 °C for These compositions are more environmentally 12 hours, shaked slowly without destroy termites friendly, but the price is relatively very expensive, body. While SGE was produced by cutting the ster- so the user community is very rare [5]. nal glands under a stereo microscope using micro Dodecenol pheromones is environmentally scissor and forceps. The severed sternal glands friendly termiticide, due to relatively cheap, ef- were then extracted using hexane as a solvent (with fective and high specify but little bit slowly in C. the composition is 4 ml hexane for 1000 gland) at 4 cynocephalus controlling. One of the efforts to °C for 12 hours. Both extracts can be used directly improve dodecenol gel performance is applying or stored at -15 °C until the next analysis process nanoparticles technology. Carrier matrix materials [7]. used in nanoparticle technology is nanometer size GC-MS analysis chitosan. It aims to cut the duration of pheromone The entire body glands and the sternal glands penetration and improve it works. were analyzed using gas chromatography fol- Chitosan in the form of nanoparticles was cho- lowed by mass spectroscopy to find out the frag- sen because it has biocompatible, biodegradable, mentation of the dominant compound in it. Anal- low toxicity and mucoadhesive properties [6]. Be- ysis GC-MS Analysis was conducted by applied Gas cause of that, the study aims to analyze the effec- Chromatograph system 6890N (Agilent Technolo- tiveness of nanoparticle of dodecenol feromone gies, Inc) combined with 5975 inert XL mass selec- body extract and sternal gland extract in con- tive detector (Agilent Technologies, Inc). Electron trolling C. cynocephalus through termite feeding Impact (EI) mass spectra was settled in the amount preference and analysis of chemical components of electron energy is 70 eV, using an HP-5MS cap- of both extracts. This research was provided bene- illary column (Agilent Technologies Inc, 0.25 mm ficial value in the development of wood production in diameter × 30 m, and 0.25 µm film thickness). technology in termite control using feeding tech- Analytic column that was used is capillary fused sil- niques. ica. Helium (He) was used as gas carrier with flow Method speed up to 37 cm/s. The samples in GC were set- This research had been done in Termite Biolo- tle in the temperature 40 °C to 220 °C in 3 °C/min. gy Laboratory, Mathematics and Science Faculty, EI mass spectra was collected from scanning the Universitas Negeri Semarang for preparing sample, flow of electron’s energy when the electron energy making dodecenol, bioassay Y and petridish assay up to 70 eV, the scanning was conducted through for about 5 months. Method used in this study was wide gaps 25 to 300 a.m.u in 0.8 s. experimental design. Dodecenol Y bioassay Termite preparation Bioassay has done under standard conditions C. cynocephalus was obtained from termite with a relative humidity of ± 70% and a room tem- farms in Entomology Laboratory Faculty of Chem- perature at 28 °C. The two extracts BE and SGE ical Product of Forest, Institute Pertanian Bo- that had been produced (dodecenol) were tested gor (Bogor, Indonesia). All termites used in this using Y bioassay on Whatmann N° 1 disc paper (15 study were adult workers/pseudergate termites cm diameter), with an intermediate angle of 120°. 6-months-old. Termites were then adapted to the In Y (3 cm) and one Y (7 cm) branch, artificial trail environment Laboratory of Termite Biology, Uni- (BE and SGE) were drawn using a micro liter syringe versitas Negeri Semarang (Semarang, Indonesia) at in which there is 1 μl extract per 1 cm marker. Oth- 28 °C and humidity 68%-70%. er extracts or pure solvents (hexane) as controls, are placed in the same conditions on the basis of Y Dodecenol pheromone production and other Y branches. Then a termite was placed in The preparation of dodecenol extract sample a small 55 mm diameter bottle at the base of Y, and was performed by liquid phase extraction method the distance traveled by the termites is measured.

Citation: Subekti N, Mar’ah R, Aisyah AN, Nada DF, Rahmawati M (2019) Nanoparticle Technology of Dodecenol as Active Components in Cryptotermes Cynocephalus Control System. Int J Nanoparticles Nanotech 5:031 Subekti et al. Int J Nanoparticles Nanotech 2019, 5:031 ISSN: 2631-5084 | • Page 3 of 6 •

The time spent between introducing termites to Ten pseudergate C. cynocephalus were placed bottles and bottle relics by termites was also mea- separately into 60 × 15 mm Petridish which there sured to determine the role of the marker in deter- was filled with 15 mm cut papers. In the control mining the behavior of termites [7]. group, the papers was moistened using 1 mL of BE and SGE were tested to 30 replicates (n = 30) distilled water, other groups separately were filled and each repetition used new termites and filter with paper, which was moistened using 1 mL of papers to prevent the effect of previous behav- nanoparticle at 6 different concentrations (0%, 5%, ioral conditioning. The activity threshold for both 10%, 15%, 20%, and 25%) and each test was per- extracts was determined as the minimum concen- formed 3 repetitions. All tests were then stored tration that induces termites to travel an average randomly in a sealed container for 4 days and the distance of more than 3 cm. While the maximum mortality was observed for each day [11]. response was 10 cm [7,8]. Result and Discussion Dodecenol nanoparticle preparation GC-MS analysis Preparation of nanoparticle was done by dis- Dodecenol is specific chemical compound pro- solving 3 grams of chitosan in 100 mL glacial ace- duced by dry wood termites, Kalotermitidae fami- tic acid 1% then homogenized at 2000 rpm for 2 ly. Dodecenol have a roll as trail following phero- hours. The homogeneous mixture was then added mone produced by pseudergates’s sternal gland. with 20 μL tween 80 0.1% and homogenized again Because of that, dodecenol can be used as an at 1000 rpm at 25 °C for 15 min. Furthermore, 200 active compound or bait in C. cynocephalus con- ml of TPP 0.1% and 2 ml BE added. This mixture was trolling system. The dodecenol existences can then homogenized at a rate of 1000 rpm for 1 hour be detected through chemical analysis of BE and and formed nanoparticles [9,10]. SGE using GC-MS (Figure 1). Figure 1 shows that Scanning Electron Microscopy (SEM) and Par- dodecenol peak of BE arises at retention time 13.05 minutes. This is appropriate to a research ticle Size Analyzer (PSA) showing that 12 carbons compound such as do- Firstly, nanoparticle formed was frozen at -25 decenol has retention time ± 13.00 minutes [12]. °C and then analyzed morphologically using SEM This mentioned that dodecenol peak had risen up (Phenom pro X Desktop SEM) at 1500× magnifi- at retention time 13.66 minutes, 13.99 minutes, cation. The droplet size of the nanoparticle was and 13.85 minutes according to its concentration. determined by photon correlation spectroscopy The chromatogram is then reinforced with mass which analyzes the fluctuations in light scattering spectra data showing the presence of dodecenol due to Brownian motion of the particles using PSA according to the peak shown in Figure 2. (Horiba LA-690 Particle Size Analyzer). Figure 2 shows that mass spectra of BE con- Petridis assay firmed the existence of dodecenol at retention time 13.05 minutes within its chemical structure

Figure 1: Chromatogram of body extract.

Figure 2: Mass spectra of body extract.

Table 1: Adaptation time and mileage of termites on ing C. cynocephalus [17]. bioassay Y (mean ± SD). Bioassay Y Extract Adaptation time (s) Distance travelled (cm) This study uses two types of extracts namely BE 355 ± 557.80 5.65 ± 0.52 Body Extract (BE) and Sternal Gland Extract (SGE). SGE 2858.5 ± 2563.98 2.15 ± 0.11 Table 1 shows that BE has more influence on termite behavior than SGE. BE affects the termite ad-

(C12H25OH). Based on the structure, dodecenol aptation time in vial bottles more quickly (355± can be included to long chain alcohol. This is sup- 557.80 s) than SGE (2858.5 ± 2563.98 s). In addi- ported by research which stated that dodecenol tion, it is also proved by termites in the BE test that has 12 carbons and can be called as lauryl alco- are able to travel a further distance of 5.65 ± 0.52 hol [13]. Lauryl alcohol is one of fatty alcohol cm compared to SGE which only affects termites to compounds such as dodecadienol, trail following walk 2.15 ± 0.11 cm. pheromone identified on Odontotermes formo- This research study show the distance traveled sanus [8]. Similarly, another study observed non- by C. cynocephalus movement on SGE bioassay adecadienone as a pheromone marker on Glos- was less than 3 cm (Table 1). It shown that SGE sotermes oculatus [14]. The pheromone is gener- pheromone is not activated as like as pheromone ally a complex of long-chain compounds that are track of Odontotermes formosanus. Inactivated highly folatile produced by certain organs. pheromone caused by the volatile dodecenol that Commonly, this pheromone is a complex of long was produced. Alcoholic pheromone such as ded- chain compound produced by specific organ called ocenol produced by C. cynocephalus has high vol- sternal gland. atility and acting as mimicking nature pheromone from Coptotermes gestroi. At the first 25 minutes Sternal gland is one of sources producing trail the activity of termites were increased, and got following pheromone in all termites species. This down slowly at the next 30 minute [18]. gland is also stated as main sources producing trail following pheromone on worker and pseudergate Sternal gland is one of the dodecenol producer of dry wood termites [15]. Trail following phero- in C. cynocephalus. Dodecenol produced by this mones found in all termites are commonly grouped organ then transported to another organ that to alcohol, aldehyde, and hydrocarbon with mo- physiologically need for regulating somebody lecular weight variations from 180 to 272 [16]. mechanisms (eg: Recognizing other worker as Termite’s pheromones are basically same for each trail pheromone). This way that dodecenol con- termite’s family, for example dodecenol as trail fol- centration in SGE was not affected to termites lowing pheromone found in Kalotermitidae includ- behavior indicated by short distance traveled. Dodecenol in C. cynocephalus was translocated

Figure 3: Decreasing of wood mass in the petridis per percent of chitosan concentration. to antenna on the termite head for recognizing vegetable insecticide in soil control (Coptotermes another termite in the colony. So that the do- sp.). Proc Seminar FMIPA UNILA 521-528. decenol concentration in the whole body extract 2. MT Ferreira, RH Scheffrahn (2011) Light attraction was higher than sternal gland extract. and subsequent colonization behaviors of alates and Petridish assay dealates of the West Indian drywood termite (Isop- tera: Kalotermitidae). Florida Entomol 94: 131-136. Y tube testing was conducted to measure the effect of pheromone in termite preferences of 3. N Subekti, T Yoshimura, F Rokhman, Z Mastur (2015) feed wood consuming. Preferences level was in- Potential for subterranean termite attack against five bamboo species in correlation with chemical dicated by mass loss of feed woods that caused by components. Proc Environ Sci 28: 783-788. termite’s wood consuming activities. Termite ap- petite was increased, when given with 0% to 10% 4. A Sumaryanto, SA Hadikusumo, GL Ndaru (2013) J of nanoparticle and decreased in the 25% to 30% Forestry Sci 7: 93. of nanoparticle concentration. Maximum wood’s 5. Y Rismayadi, Arinana (2009) J Sci Technol Forest mass loss up to 13.86% in 10% of nanoparticle Product 2: 32. group. Then, the wood that submerged in the 6. A Rahayu A (2014) Formulasi Tablet dari Nanopartikel 20% of nanoparticle also loss high amount up to Ekstrak Daun Sambiloto (Andrographis paniculata 13.80% but not significantly different. It can be [Burn.f.] Ness) dengan Metode Cetak Langsung, conclude that usage of 10% nanoparticle is op- Faculty of Pharmacy, Jakarta, Indonesia. timum and effective dose in the C. cynocephalus 7. A Peppuy, A Robert, E Semon, C Ginies, M Lettere, control system (Figure 3). et al. (2001) (Z)-dodec-3-en-1-ol, a novel termite Acknowledgment trail pheromone identified after solid phase microextraction from Macrotermes annandalei. J Acknowledgment of the authors convey to the Insect Physiol 47: 445-453. Directorate General of Learning and Student Af- fairs, Ministry of Research, Technology and Higher 8. P Wen, BZ Ji, DS Dusses (2014) Trail communication Education, Indonesia with contract number 547/ regulated by two trail pheromone components in the B3.1/KM/2017 dated March 09, 2017. Thank you fungus-growing termite Odontotermes formosanus also the author to convey to the Universitas Negeri (Shiraki). PLOS ONE 9: 90906. Semarang in Indonesia and all parties who are will- 9. SW Ali, S Rajendran, M Joshi (2011) Synthesis and ing to assist in this research. characterization of chitosan and silver loaded chitosan nanoparticles for bioactive polyester. References Carbohydrate Polym 83: 438-446. 1. Zulyusri, Desyanti, U Mardia (2013) Effectiveness 10. L Zhao, L Shi, Z Zhang, J Chen, D Shi, et al. (2011) of seed leaves (Sambucus javanica Reinw) as a Preparation and application of chitosan nanoparticles

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