, for the identification of efficacy of ?-caryophyllene 122 oxide, the safety evaluation showed no prohibited or restricted components. For safety reasons, 123 the in vitro 3T3 NRU phototoxicity test, 2004.
, N° 130 P6110401, Lot. 09I006), low passage number (<50)
, Minimum Essential Medium, PAN BIOTECH. lot 1874561) supplemented with penicillin 100
At the end of the incubation period, the culture medium was decanted 136 and replaced by 100 µL of Hank's balanced salt solution (HBSS) containing the appropriate 137 concentrations of the test substance (eight different concentrations). Subsequently, cells were 138 incubated at 37°C (5% CO 2 ) in the dark for 60 min. From the two plates prepared for each 139 series of test substance concentrations and the controls, one was randomly selected for the 140 determination of cytotoxicity without irradiation (-Irr), and the other for the determination of 141 photo-cytotoxicity with irradiation (+Irr), The irradiation procedure was performed using a 142 solar simulator Suntest CPS+ apparatus (Atlas Material Testing Technology BV ,
France) equipped with a xenon arc lamp (1,100 W), a special glass filter restricting 144 transmission of light below 290 nm and a near IR-blocking filter. The irradiance was fixed at ,
, The Neutral Red medium was removed and destaining solution (50% ethanol, 1% acetic 152 acid, 49% distilled water; 50 µL per well) was added into each well. Then, the plates were
The mean OD value of blank wells (containing only Neutral Red desorbed 165 solution) was subtracted from the mean OD value of three treated wells (dilutions of the test 166 material, positive control or HBSS). The percentages of cell viability were calculated as: 313 has focused on potent synthetic compounds, Neutral Red without irradiation (IC 50 -Irr) and with irradiation, vol.314, p.320, 2009. ,
, Several plant-based essential oils have been evaluated for mosquito repellent activity 321 as protection against mosquitoes and other arthropod pests in Thailand
Vetiveria 323 zizanioides (L.) Nash (Vetiver), Syzygium aromaticum (L.) Merr. & L.M. Perry (Clove flower), and Zingiber 325 officinale Roscoe (Ginger), vol.324, 2014. ,
, with insect behavior and growth, 2013.
The plant products have been effective as insect repellents or 330 insecticidal agents and one of the potential repellent compounds is ?-caryophyllene oxide 331, 2018. ,
, , 2005.
Strong repellency against An. gambiae was also reported from a combination of 339 linalool, ?-caryophyllene oxide, ?-terpinene, and 1-methylpyrrole (45:39:8:8), and essential oil 340 of Croton pseudopulchellus Pax, Bhandari oil, obtained from leave parts, contains ?-caryophyllene oxide as one of its major 337 constituents and displays strong repellent activity against females of An. gambiae (Suleiman et 338, 2005. ,
Administration (FDA) and the European Food Safety Authority (EFSA, 2016. ,
, The two most common plant-associated repellent compounds are p-menthane-3,8-diol (derived 347 from the Australian lemon-scented gum tree) and picaridin (a synthetic derivative of pepper)
The phototoxicity results obtained using the in vitro in mammalian cells. Likewise, the current study showed that ?-caryophyllene oxide 358 did not induce genotoxicity at the chromosomal level, 348 these have been tested for toxicity, 2009. ,
Their results showed that the flavoring agent was 361 devoid of mutagenic effects, both at the gene level (frameshift or based-substitution mutations) 362 and on chromosomes, 2013. ,
2017) reported that essential oils of citronella, vetiver, hairy basil, 366 and kaffir lime had good irritant and repellent effects on mosquito vectors compared to DEET. study showed that at higher concentrations (0.5 and 1.0%), ?-caryophyllene 371 oxide had significantly greater repellent and irritant effects compared to DEET, 2016. ,
, The results in contact and non-contact trials indicated that escape responses of 377 mosquitoes to the ?-caryophyllene oxide were significantly greater than with DEET, similar to 378 the previous study by, 2019.
, An. minimus presented the highest sensitivity to both types of escape responses (contact 386 irritancy and noncontact-spatial repellency) at 0.5-1% (v/v)
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NC=non-contact; Esp= escaped mosquitoes; NE= Non escape mosquitoes ,
, § Positive controls: mitomycin C (0.05 µg.mL -1 ) without S9 mix and benzo
, mL -1 ) with S9 mix; MNC: Micronucleated cells per 1,000; P: Probability of the comparison 637 between the negative control and the tested dose using the Chi-squared test
, NS: non-significant activity
, 25%: (A) Ae. to ?-caryophyllene oxide and DEET at 1%: (A) Ae. albopictus, 655 (B) An. dirus, DEET at 0