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Composition and Toxicity of Chinese Dracocephalum Moldavica (Labiatae) Essential Oil Against Two Grain Storage Insects

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Composition and Toxicity of Chinese Dracocephalum Moldavica (Labiatae) Essential Oil Against Two Grain Storage Insects Journal of Medicinal Plants Research Vol. 5(18), pp. 4621-4626, 16 September, 2011 Available online at http://www.academicjournals.org/JMPR ISSN 1996-0875 ©2011 Academic Journals Full Length Research Paper Composition and toxicity of Chinese Dracocephalum moldavica (Labiatae) essential oil against two grain storage insects Sha Sha Chu 1, Shao Liang Liu 2, Qi Zhi Liu 1, Zhi Long Liu 1* and Shu Shan Du 3 1Department of Entomology, China Agricultural University, 2 Yuanmingyuan West Road, Haidian District, Beijing 100094, PR China. 2Department of Biology, Faculty of Preclinical Medicine, GuangXi Traditional Chinese Medical University, Nanning 530001, PR China. 3State Key Laboratory of Earth Surface Processes and Resource Ecology, Beijing Normal University, Beijing 100875, China. Accepted 5 July, 2011 Essential oil of Dracocephalum moldavica flowering aerial parts was extracted via hydrodistillation. A total of 38 components of the essential oil were identified. 1, 8-Cineol (31.25%) and 4-terpineol (22.82%) were the two main components of the essential oil followed by cumin alcohol (4.29%) and α-terpineol (4.21%). The essential oil exhibited strong fumigant toxicity against Sitophilus zeamais and Tribolium castaneum adults with LC 50 values of 2.65 and 0.88 mg/L, respectively. The essential oil also showed contact toxicity against S. zeamais and T. castaneum adults with LD 50 values of 22.10 and 18.28 µµµg/adult, respectively. The essential oil of D. moldavica may have the potential to be developed as a new natural fumigant/insecticide for the control stored product insects. Key words: Dracocephalum moldavica , Sitophilus zeamais , Tribolium castaneum , contact toxicity, essential oil composition. INTRODUCTION Sitophilus and Tribolium species are two of the major alternatives with fumigant action. Plant essential oils and pests of stored grains and grain products in the tropics their components have been shown to possess potential and subtropics (Liu and Ho, 1999). Infestations not only to be developed as new fumigants and they may have cause significant losses due to the consumption of the advantage over conventional fumigants in terms of grains; they also result in elevated temperature and low mammalian toxicity, rapid degradation and local moisture conditions that lead to an accelerated growth of availability (Isman, 2006). Essential oils derived from molds, including toxigenic species (Magan et al., 2003). more than 75 plant species have been evaluated for Control of the stored product insects is based on the fumigant toxicity against stored product insects so far application of synthetic insecticides/fumigants. However, (Rajendran and Srianjini, 2008). repeated use of those fumigants for decades has led to During our screening program for new agrochemicals resurgence of stored-product insect pests, sometimes from Chinese medicinal herbs and local wild plants, resulted in the development of resistance, and had essential oil of Dracocephalum moldavica L. (Family: undesirable effects on non-target organisms (United Labiatae) flowering aerial parts was found to possess States Environmental Protection Agency, 1993; Zettler insecticidal toxicity to the maize weevils, Sitophilus and Arthur, 2000). These problems have highlighted the zeamais Motschulsky and the red back flour beetles, need to develop new types of selective insect-control Tribolium castaneum (Herbst). D. moldavica is an annual herbaceous aromatic plant and native to central Asia and is naturalized in eastern and central Europe (Tian et al., 2009). In China, it is predominantly found in the north of *Corresponding author. E-mail: [email protected]. Tel: the country. This plant has been of interest to Uygur +86-10-62732800. Fax: +86-10-62732800. traditional medicine, especially in north Xinjiang 4622 J. Med. Plant. Res. automonous region. It is used as a food ingredient, as a flour mixed with yeast (10:1, w/w) while the maize weevils were tea, as an herbal drug for its reputed medicinal reared on whole wheat at 12 to 13% moisture content in glass jars properties, for example, the treatment of stomach and (diameter 85 mm, height 130 mm) at 29 to 30°C and 70 to 80% relative humidity. Unsexed adult weevils and beetles used in all the liver disorders, headaches and congestions, with strong experiments were about 2 weeks old. cardioprotective effect (Chinese Pharmacopoeia Commission, 1999). Previous phytochemical studies on D. moldavica Gas chromatography-mass spectrometry resulted in the identification of phenolics (rosmarinic acid, The essential oil of D. moldavica flowering aerial parts was caffeic acid), iridoids, sesquiterpenoids, flavonoids, subjected to GC-MS analysis on an Agilent system consisting of a triterpenoids, and steroids (Gu et al., 2004; 2005; model 6890N gas chromatograph, a model 5973N mass selective Dastmalchi et al., 2006, 2007; Feng and Li, 2006; Popova detector (EIMS, electron energy, 70 eV), and an Agilent et al., 2008; Sultan et al., 2008). Zeng et al. (2010) ChemStation data system. The GC column was an HP-5ms fused reviewed the constituents derived from the plants of silica capillary with a 5% phenyl-methylpolysiloxane stationary genus Dracocephalum (including terpenoids, flavonoids, phase, film thickness of 0.25 µm, a length of 30 m, and an internal diameter of 0.25 mm. The GC settings were as follows: the initial alkaloids, lignans, phenols, coumarins, and cyanogenic oven temperature was held at 60°C for 1 min and ramped at 10°C glucosides) and the biological activities were also min −1 to 180°C held for 1 min, and then ramped at 20°C min −1 to discussed . The composition of the essential oil of the 280°C and held for 15 min. The injector temperature was aerial flowering parts of D. moldavica has been maintained at 270°C. The sample (1 µl) was injected neat, with a split ratio of 1: 10. The carrier gas was helium at flow rate of 1.0 ml investigated previously (Holm et al., 1988; Shatar and −1 −1 Altantsetseg, 2000; El-Baky and El-Baroty, 2008; Sonboli min . Spectra were scanned from 20 to 550 m/z at 2 scans s . Most constituents were identified by gas chromatography by et al., 2008). The essential oil of D. moldavica was comparison of their retention indices with those of the literature (El- evaluated as a seed protectant against the pulse beetle Baky and El-Baroty, 2008; Sonboli et al., 2008) or with those of (Callosobruchus maculatus ) (Aziz and Abbass, 2010). authentic compounds available in our laboratories. The retention However, no reports on toxicity of D. moldavica essential indices were determined in relation to a homologous series of n- oil against the two grain storage insects were available so alkanes (C 8 to C24 ) under the same operating conditions. Further identification was made by comparison of their mass spectra with far. This study analyses the chemical composition and those stored in NIST 08 and Wiley 275 libraries or with mass toxicity of essential oil of D. moldavica against the two spectra from literature (Adams, 2007). Component relative grain storage insects. percentages were calculated based on normalization method without using correction factors. MATERIALS AND METHODS Contact toxicity Plant material Range-finding studies were run to determine the appropriate testing concentrations of the essential oil. A serial dilution (4.0 to 10.0% v/v Fresh aerial parts (10 kg of leaves, stems and flowers) of D. for S. zeamais , 0.5 to 5.0% v/v for T. castaneum , six moldavica were harvested in August 2009 from Xiaolongmeng concentrations) of the essential oil was prepared in n-hexane. National Forest Park (Mentougou District, Beijing 102300). The Aliquots of 0.5 µl per insect were topically applied dorsally to the aerial parts were air-dried for one week and ground to a powder thorax of insects, using a Burkard Arnold microapplicator. Controls using a grinding mill (Retsch Mühle, Germany). The species was were determined using 0.5 µl n-hexane per insect. Ten insects were identified by Dr. Liu, Q.R. (College of Life Sciences, Beijing Normal used for each concentration and control, and the experiment was University, Beijing 100875, China), and a voucher specimen (BNU- replicated six times. Both treated and control insects were then zhilongliu-2009-08-29-031) was deposited at the Herbarium (BNU) transferred to glass vials (10 insects/vial) with culture media and of College of Life Sciences, Beijing Normal University. The aerial kept in incubators at 29 to 30°C and 70 to 80% relative humidity. parts were ground to a powder using a grinding mill (Retsch Mühle, Mortality was observed daily until end-point mortality was reached Germany). Each 600 g portion of powder ground was mixed in 1800 one week after treatment. Results from all replicates were subjected ml of distilled water and soaked for 3 h. The mixture was then to probit analysis using the PriProbit program V1.6.3 to determine boiled in a round-bottom flask, and steam distilled for 6 to 8 h. LD 50 values (Sakuma, 1998). Volatile essential oil from distillation was collected in a flask. Separation of the essential oil from the aqueous layer was done in a separatory funnel, using the non-polar solvent, n-hexane. The Fumigant toxicity solvent was evaporated using a vacuum rotary evaporator (BUCHI Rotavapor R-124, Switzerland). The sample was dried over A Whatman filter paper (diameter 2.0 cm, CAT No. 1001020) was anhydrous Na 2SO 4 and kept in a refrigerator (4°C) for subsequent placed on the underside of the screw cap of a glass vial (diameter experiments. 2.5 cm, height 5.5 cm, volume 24 ml). Range-finding studies were run to determine the appropriate testing concentrations of the essential oil. Ten microliters of 0.2 to 12.0% of essential oil (v:v) Insects was added to the filter paper. The solvent was allowed to evaporate for 30 s before the cap was placed tightly on the glass vial (with 10 The maize weevils ( S.
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