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Chemical Constituents and Insecticidal Activity of the Essential Oils Extracted from Artemisia giraldii and Artemisia rubripes against Two Stored Product Insects

Jun Yu Liang1,2, Xue Ting Liu1, Jin Gu1, Yan Liu1, Xiao Yan Ma1, Nan Lv1, Shan Shan Guo2, Jun Long Wang1, Shu Shan Du2* and Ji Zhang1

1College of Life Science, Northwest Normal University, Lanzhou, PR China

2Beijing Key Laboratory of Traditional Chinese Medicine Protection and Utilization, Beijing Normal University, Beijing, PR China

*Corresponding Author:
Shu Shan Du
Beijing Key Laboratory of Traditional
Chinese Medicine Protection and Utilization, Beijing
Normal University, Haidian District, Beijing 100875, PR China
Tel: +861062208022
E-mail: [email protected]

Received date: August 05, 2016; Accepted date: August 22, 2016; Published date:August 25, 2016

Citation: Liang JY, Liu XT, Gu J, Liu Y, Ma XY, et al. (2016) Chemical Constituents and Insecticidal Activity of the Essential Oils Extracted from Artemisia giraldii and Artemisia rubripes against Two Stored Product Insects. Med Chem (Los Angeles) 6:541-545. doi:10.4172/2161-0444.1000396

Copyright: © 2016 Liang JY, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Abstract

The chemical constituents of the essential oils extracted from Artemisia giraldii and Artemisia rubripes were analyzed by GC-MS. The bioactivities of two essential oils against Tribolium castaneum and Lasioderma serricorne were investigated. The main constituents in A. giraldii essential oil were 1,8-cineole (40.72%), camphor (22.50%), terpinen-4-ol (12.41%) and α-terpineol (4.14%). β-Farnesene (12.23%), 1,8-cineole (11.09%), β-caryophyllene (10.68%), germacrene D (10.01%), camphor (7.01%) were principal constituents in A. rubripes essential oil. The essential oil of A. giraldii possessed contact and fumigant toxicities (LD50=23.72 μg/adult and LC50=16.78 mg/l air, respectively) against T. castaneum adults. In addition, the essential oil of A. giraldii possessed contact and fumigant toxicities (LD50=8.88 μg/adult and LD50=5.41 mg/l air, respectively) against L. serricorne adults. The essential oil of A. rubripes showed contact and fumigant toxicities (LD50=16.18 μg/adult and LD50=25.10 mg/l air, respectively) against L. serricorne adults, and the very low contact and fumigant toxicities against T. castaneum. The results indicated that the essential oil of A. giraldii had the potential to be developed as insecticide for control of T. castaneum and L. serricorne.

Keywords

Artemisia rubripes; Artemisia giraldii; Tribolium castaneum; Lasioderma serricorne; Contact toxicity; Fumigant toxicity

Introduction

Long-time and excess usage of synthetic chemical pesticides have caused many direct and indirect harm to environments and non-target organisms, although these pesticides play very important roles in increasing grain yield and protecting store grain against pest over past more than half century [1-3]. The ecological balance would be broken down if pest are eliminated completely [4]. Therefore, there is urgent need to develop a safe and alternative insecticide that has a potential to replace the toxic insecticides. The essential oils (EOs) are focused of much attention for their characters of being obtained easily, low-toxicity, eco-friendly, being degraded quickly [5]. EOs had been used long ago around the world, and now they have been made into various products for medicine, food protection, cosmetic [6], thus, EOs are relatively safe. Since the artemisinin from Artemisia annua was found and used to fight against malaria, manyplants of Artemisia spp. have been used as important resource of artemisinin [7,8]. The studies on the artemisinin and it’s derivative make great contributions to the human’s health. These achievements have excited researchers to treat diseases by natural occurring substances. In addition, these materials can be used in other fields, such as food, cosmetics, and insecticide. The bioactivities of many Artemisia spp. plants against some pest have been investigated, and EOs extracted from these plants have shown obvious insecticidal or repellent activities [9-13]. The contact toxicity is a kind of common insecticidal model When the activity material come into contact the insects, the material enter the interior of insect and cause the death of insects.

Artemisia giraldii Pamp. and Artemisia rubripes Nakai are two species from Artemisia spp., and they are rich in resources in China [14]. In the previous studies, the essential oil of Artemisia giraldii showed the contact and fumigant toxicity against Sitophilus zeamais adults [15]. A literature survey showed that there are no reports on the insecticidal activity of the essential oils extracted from A. giraldii andA. rubripes against Tribolium castaneum and Lasioderma serricorne.

The red flour beetle T. castaneum (Herbst) (Coleoptera: Tenebrionidae) and the cigarette beetle L. serricorne (Fabricius) (Coleoptera: Anobiidae) are two most common species of the insects in stored food. The secreta of T. castaneum can cause flour to agglomerate, change in color, and going bad, and the secreta contain benzoquinone that is a kind of carcinogenic substance. L. serricorne cause damage to store grains and tobacco [16]. These two species of insects have the characteristics of rapid reproduction, strong adaptation, and worldwide distribution, so they cause significant damage to the storage of many crops and products and great economic loss [17].

In this work, the insecticidal activities of the essential oil extracted from A. giraldii and A. rubripes againstT. castaneum and L. serricorne were investigated.

Materials and Methods

Plant material

The aerial part of A. giraldii andA. rubripes were gathered from Tianshui city (34°34′03″N,105°42′58″E, altitude 1244 m) and Tanchang county (33°58′01″N,104°25′15″E, altitude 1776 m), respectively, Gansu Province, China in June 2015. The species were identified by Dr. Liu, Q.R. and the voucher specimen of A. giraldii (BNU-dushushan-2015072401) and A. rubripes (BNU-dushushan-2015072002) were deposited at the Herbarium (BNU) of College of Resources Science and Technology, Beijing Normal University.

Insect

T. castaneum and L. serricorne were feed in constant temperature oven at 29 ± 1°C and 70-80% relative humidity and the fodder was mixture of wheat flour and yeast (10:1, w/w). The unsexed insects in all of the experiments were 1-2 weeks old adults. All experiments were carried out in constant temperature oven that have same condition with insect feed.

Extraction and analysis of essential oil

The aerial part of two plants were grounded into powder after airdried respectively. The powders were weighed and transferred into the steam distillation equipment, and the hydrodistillation was carried out for 6 h to get crude essential oils of two plants, then anhydrous sodium sulphate was used to remove extra water. The volumes of waterless essential oils were recorded and the yield was calculated. Two waterless essential oils were the test sample of insecticidal activities and were stored at 0-4°C. The chemical constituents of essential oils of two plants were analyzed on a GC-MS instrument (Agilent 6890N - Agilent 5973N). The detector of GC instrument was flame ionization detector (FID). Capillary column was HP-5MS (30 m × 0.25 mm × 0.25 μm). The temperature programming of GC instrument was original oven temperature at 60°C for keeping 1 min and risen at 10°C/min to 180°C for keeping 1 min, and then risen at 20°C/min to 280°C for keeping 15 min. The injector temperature was maintained at 270°C. The samples (1 μl, dilute to 1% with hexane) were injected with a split ratio of 1:10. Helium was used as carrier gas and the flow rate was 1 ml/ min. The retention indices (RI) were calculated by using a homologous series of n-alkanes (C5-C36) under the same operating conditions. The constituents were identified by comparing their mass spectra and calculating RI values with those in NIST 05 and Wiley 275 libraries, and those published in the literatures [18-22]. Relative percentages of the individual constituents of the essential oils were obtained by averaging the GC-FID peak area% reports.

Contact toxicity

A bioassay of contact toxicity of two essential oils were designed to determine 50% lethal doses (LD50) based on the method described by Liu and Ho [23] against T. castaneum and L. serricorne. The essential oil was dissolved in n-hexane to prepare a serial testing solution. A liquots of 0.5 μl of the dilutions were applied to the dorsal thorax of T. castaneum and L. serricorne. n-Hexane solvent and pyrethrins (pyrethrin I and II, 37%, was purchased from Dr. Ehrenstorfer GmbH) were used as the negative and positive control, respectively. Five replicates were done for all tests and ten insects were used in each replication. Then the insects were placed in constant temperature oven at 29 ± 1°C and 70- 80% relative humidity. The dead insects were counted after 24 h. The insects were considered dead when no leg or antennal movements were observed by touching them. The data were corrected for control mortality using Abbott's formula and the LD50 values were calculated by using Probit analysis (IBM SPSS V20.0) [24].

Fumigant toxicity

The fumigant activity of the essential oil against two species of insects was tested as described by Liu and Ho [23]. A serial dilution of the essential oils was prepared in n-hexane. A Whatman filter paper (diameter 2.0 cm) was infiltrated with 10 μl dilution, and then placed on the underside of the screw cap of a glass vial (diameter 2.5 cm, height 5.5 cm, volume 25 ml). The solvent was evaporated for 20 s to remove n-hexane before the cap was placed tightly on the glass vial. Each of glass vial contained 10 insects inside to form a sealed chamber. n-Hexane was used as a negative control. The MeBr and phosphine were used as positive control against T. castaneum and L. serricorne respectively. Five replicates were operated for all tests. The dead insects were counted after 24 h. The insects were considered dead when no leg or antennal movements were observed by touching them. The data were corrected for control mortality using Abbott's formula. The 50% lethal concentration (LC50) values were calculated by using Probit analysis (IBM SPSS V20.0) [24].

Results and Discussion

Chemical composition of the essential oil

The yield of the essential oils of A. giraldii and A. rubripes were 0.45% (v/w) and 0.50% (v/w) respectively. By GC-MS analysis, the main constituents of essential oil of A. giraldii were 1,8-cineole (40.72%), camphor (22.50%), terpinen-4-ol (12.41%), α-terpineol (4.14%) and 8-cedren-13-ol (3.60%). The main constituents of essential oil of A. rubripes were β-farnesene (12.23%), 1,8-cineole (11.09%), β-caryophyllene (10.68%), germacrene D (10.01%), camphor (7.01%) . The constituents of two essential oils were listed in Tables 1 and 2.

PeakNo. RIa) Compound Chemicalformula Composition(%)
1 1017 1,8-Cineole C10H18O 40.72
2 1060 γ-Terpinene C10H16 1.83
3 1185 cis-β-Terpineol C10H18O 1.59
4 1118 cis-p-Menth-2-en-1-ol C10H18O 1.97
5 1120 Camphor C10H16O 22.50
6 1147 Terpinen-4-ol C10H18O 12.41
7 1174 α-Terpineol C10H18O 3.60
8 1226 cis-Carveol C10H16O 0.37
9 1230 Carvone C10H14O 0.26
10 1350 (+)-α-Longipinene C15H24 0.15
11 1372 (-)-α-Copaene C15H24 0.21
12 1414 β-Caryophyllene C15H24 1.36
13 1456 Z,Z,Z-1,5,9,9-tetramethyl-1,4,7,-Cycloundecatriene C15H24 0.58
14 1479 GermacreneD C15H24 2.28
15 1487 (-)-α-Selinene C15H24 0.36
16 1561 GermacreneB C15H24 0.40
17 1563 Spathulenol C15H24O 0.35
18 1566 Caryophylleneoxide C15H24O 0.32
19 1669 8-Cedren-13-ol C15H24O 4.15
20 1721 5,11-Guaiadiene C15H24 1.24
    Monoterpenoids   85.25
    Sesquiterpenoids   11.4
    Total   96.65

Table 1: Chemical constituents of the essential oil derived from A. giraldii.

PeakNo. RIa) Compound Chemicalformula Composition(%)
1 1017 1,8-Cineole C10H18O 11.09
2 1114 Thujone C10H16O 1.71
3 1120 Camphor C10H16O 7.01
4 1147 α-Pinocarvone C10H14O 3.79
5 1159 Borneol C10H18O 2.94
6 1179 Terpinen-4-ol C10H18O 2.46
7 1344 α-Cubebene C15H24 0.83
8 1411 α-Gurjunene C15H24 0.72
9 1414 β-Caryophyllene C15H24 10.68
10 1450 β-Farnesene C15H24 12.23
11 1456 Z,Z,Z-1,5,9,9-tetramethyl-1,4,7,-Cycloundecatriene C15H24 2.59
12 1462 Di-epi-α-cedrene C15H24 5.73
13 1472 α-Curcumene C15H22 4.64
14 1479 GermacreneD C15H24 10.01
15 1489 Bicyclogermacrene C15H24 1.57
16 1510 δ-Cadinene C15H24 2.60
17 1553 Davanone C15H24O2 2.80
18 1566 Caryophylleneoxide C15H24O 1.48
19 1578 Ledol C15H26O 1.71
20 1612 (-)-T-Muurolol C15H26O 1.94
21 1621 (-)-α-Cadinol C15H26O 1.49
    Monoterpenoids   29.00
    Sesquiterpenoids   61.02
    Total   90.02

Table 2: Chemical constituents of the essential oil derived from A. rubripes.

In this investigation, 1,8-cineole and camphor could be detected in the essential oils of two plants, but content of the two compounds were different in essential oil of A. giraldii and A. rubripes. The constituents of A. giraldii and A. rubripes essential oils were different from literatures reported. For example, literature has showed main constituents in A. giraldii essential oil are β-pinene (13.18%), iso-elemicin (10.08%) [15]. The main components of A. rubripes essential oil were camphor (26.94%), 1,8-cineole (15.59%), caryophyllene (13.29%) [25]. These differences might be caused by the variety of harvest locale, time, altitude and local climatic. Further studies on standardization are needed for studying the essential oil into insecticides.

Contact toxicity

The contact toxicity of the essential oil of A. giraldii and A. rubripes against T. castaneum and L. serricorne were listed in Table 3. The results exhibited the essential oil of A. giraldii possessed contact toxicity (LD50=23.72 μg/adult and LD50=8.88 μg/adult) against T. castaneum and L. serricorne, respectly. The essential oil of A. rubripes showed contact toxicity (LD50=16.18 μg/adult) against L. serricorne, but it showed the lower contact toxicity (LD50=150.92 μg/adult) against T. castaneum. Under the same test method, the essential oil of A. giraldii showed more susceptible contact toxicity than A. rubripes against T. castaneum.

Insect Treatment LD50(95%FL)(µg/adult) Slope±SE Chisquare(χ2) p-value
TC A.giraldii 23.72(21.49-26.27) 4.47±0.47 12.34 0.965
A.rubripes 150.92(137.89-165.92) 5.43±0.66 6.25 0.920
Pyrethrinsa 0.26(0.22-0.30) 3.34±0.32 13.11 0.925
LS A.giraldii 8.88(8.04-9.80) 4.35±0.41 11.26 0.998
A.rubripes 16.18(14.53-18.00) 3.82±0.37 10.99 0.998
Pyrethrinsa 0.24(0.16-0.35) 1.31±0.20 17.36 0.916

Table 3: Contact toxicity of essential oil of A. giraldii and A. rubripes against T. castaneum (TC) and L. serricorne (LS) adults.

1,8-Cineole, camphor and terpinen-4-ol were main constituents of the essential oil of A. giraldii. In the same test methods and conditions, 1,8-cineole (LD50=16.18 μg/adult) [26] and terpinen-4-ol (LD50=16.18 μg/adult) [27] possessed same level of contact activities with essential oil against T. castaneum, but camphor (LD50=16.18 μg/adult) [13] showed more low contact toxicity than the essential oil of A. giraldii against T. castaneum. Terpinen-4-ol (LD50=5.4 μg/adult) [28] showed more obvious contact activity than essential oil of A. giraldii against L. serricorne, but 1,8-cineole (LD50=15.58 μg/adult) [26] and camphor (LD50=13.44 μg/adult) [28] showed lower contact activity. It indicated 1,8-cineole and terpinen-4-ol played same functions for contact activity of essential oil of A. giraldii against T. castaneum, and terpinen-4-ol was more key components than 1,8-cineole for contact activity of essential oil of A. giraldii against L. serricorne.

The reason of the very low contact toxicity of the essential oil of A. rubripes against T. castaneum may be related to the low content of 1,8-cineole, camphor and terpinen-4-ol in the essential oil. But the essential oil of A. rubripes showed obvious contact toxicity against L. serricorne. It indicated 1,8-cineole, camphor and terpinen-4-ol may be not key factors in the essential oil of A. rubripes against L. serricorne.

Fumigant toxicity

The fumigant toxicity of the essential oils of A. giraldii and A. rubripes against T. castaneum and L. serricorne were listed in Table 4. The essential oil of A. giraldii show fumigant toxicity (LD50=16.78 mg/l air and LD50=5.41 mg/l air) against T. castaneum and L. serricorne, respectly. The essential oil of A. rubripes showed fumigant toxicity (LD50=25.10 mg/l air) against L. serricorne and very low the fumigant toxicity (LD50=97.78 mg/l air) against T. castaneum. Under the same test method, the essential oil of A. giraldii showed more susceptible fumigant toxicity than A. rubripes against two species of insects.

Insect Treatment LC50(95%FL)(mg/lair) Slope±SE Chisquare(χ2) p-value
TC A.giraldii 16.78(15.29-18.64) 4.68±0.44 8.06 0.998
A.rubripes 97.78(88.60-108.17) 4.51±0.48 8.13 0.990
MeBr 1.05 8.36±0.91 12.21 0.890
LS A.giraldii 5.41(5.21-5.63) 13.12±1.38 12.31 0.965
A.rubripes 25.10(23.12-27.20) 6.32±0.67 5.04 0.984
Phosphinea 9.23×10-3(7.13×10-3-11.37×10-3) 2.12±0.27 11.96 -

Table 4: Fumigant toxicity of essential oils of A. giraldii and A. rubripes against T. castaneum (TC) and L. serricorne (LS) adults.

The main components (1,8-cineole and terpinen-4-ol) of essential oil of A. giraldii showed obvious fumigant toxicity (LD50=5.47 mg/l air and LD50=3.7 mg/l air) [26,27] against T. castaneum and fumigant toxicity (LD50=5.18 mg/l air and LD50=1.3 mg/l air) [26,27] against L. serricorne. The fumigant toxicity of camphor has not been observed against T. castaneum [13], but the camphor showed stronger fumigant toxicity (LD50=2.36 mg/l air) [28] against L. serricorne. 1,8-cineole and terpinen-4-ol possessed more stronger fumigant activity than the essential oil of A. giraldi against T. castaneum. Moreover 1,8-cineole showed same level of fumigant activity with essential oil of A. giraldi, but terpinen-4-ol and camphor possessed more obvious fumigant activity than essential oil of A. giraldi against L. serricorne. It showed that the fumigant activity of essential oil of A. giraldi were related to the main components of 1,8-cineole and terpinen-4-ol, and terpinen- 4-ol played more important role for contribution of fumigant activity against T. castaneum and L. serricorne.

The fumigant toxicity of the essential oil of A. rubripes against T. castaneum was very low. The reason may be due to the content of 1,8-cineole, camphor and terpinen-4-ol were low in the essential oil of A. rubripes.

The secondary metabolites of plant affect biochemical processes thus product insecticidal result by poisoning nervous system or regulating growth process of insects [29]. It were reported that the main reason of EOs and their constituents can product the insecticidal effect was due to impact the insect’s some targets of nervous system. For example, some EOs acted by inhibiting the activity of AChE which is one of main resistance mechanism in insect pests [30-32]. Also, blockage of the GABA-gated chloride channel reduces neuronal inhibition, which leads the insects to overexcite thus cause convulsions, and death [33]. Thujone and thymol are two common insecticidal components of EOs, and it has been confirmed that they acts on GABA receptors [34]. On the other hand, the researchers found that some constituents of EOs (eugenol or thymol) may work by blocking octopamine receptors [35].

1,8-cineole, camphor and terpinen-4-ol were main components of the essential oil extracted from A. giraldii, and the total content of three components exceed 75 percent. These components were found to inhibit the activity of AChE [30,36]. 1,8-Cineole and camphor showed certain toxicity when individually applied, but binary mixtures were synergistic insecticidal bioassays against cabbage looper [37]. 1,8-cineole can stimulate the response of pheromone sensitive sensilla of Periplaneta americana by act on octopamine receptor [38]. 1,8-Cineole and camphor both showed significant effects on the GABA receptors of insect [39].

In conclusion, the insecticidal activity of essential oil extracted from A. giraldii may be due to synergistic action of three main componengs. The several mechanisms together affect multiple targets of insects, thereby impacting more effectively cellular activity and biological processes of insects.

Conclusions

The investigation indicated that the essential oil of A. giraldii has potential as an alternative to synthetic pesticides against T. castaneum and L. serricorne, and essential oil of A. rubripes can turn into novel production of pest management against L. serricorne. These essential oils reducing the adverse effects on the health of users and consumers. They are economical and their environmental impact is low.

Some components isolated from natural materials possess good insecticidal activity, but they also have some side effects. For example, pyrethrins and nicotinyl insecticides have showed toxicity to many species, including birds, fishes, and bees [40]. Thus, blindly pursuit pure components to control pest could be inappropriate. The biological activities of natural products are related to functional groups, rather than only one component, and, some mixture components from natural materials possess insecticidal activity and low toxicity to non-target organisms. Thus, the mixture components from natural materials could not only have insecticidal activities but also have little risk on ecology balance. EOs have the dual-advantages mentioned above, although its insecticidal effect might be weaker than that of the synthetic chemical pesticides.

Conflict of Interest

The authors declare no conflict of interest.

Acknowledgements

This project was supported by the Beijing Municipal Natural Science Foundation (No. 7142093), Natural Science Foundation of China (No. 21365019) and Fundamental Research Funds for the Central Universities. The authors thank Dr. Liu Q.R. from College of Life Sciences, Beijing Normal University, Beijing 100875, for the identification of the investigated medicinal herb.

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