Reference : Harmonia axyridis (Pallas) secondary metabolites quantification in relation with apos...
Scientific congresses and symposiums : Poster
Life sciences : Multidisciplinary, general & others
Harmonia axyridis (Pallas) secondary metabolites quantification in relation with aposematism : Part I : Harmonine quantification
Fischer, Christophe mailto [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Heuskin, Stéphanie mailto [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Laurent, Pascal mailto [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Marlet, Christelle [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Durieux, Delphine mailto [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Verheggen, François mailto [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Lognay, Georges mailto [Université de Liège - ULiège > > Gembloux Agro-Bio Tech >]
Eleventh International Symposium on Hyphenated Techniques in Chromatography (HTC-11)
du 27 janvier 2010 au 29 janvier 2010
Catholic University College Bruges-Ostend (KHBO)
[en] Harmonia axyridis ; Coccinellidae ; Alkaloïd ; Fast-GC ; Aposematism ; Validation ; Harmonine
[en] These last few years, the multicoloured Asian ladybird, Harmonia axyridis Pallas (Insects: Coleoptera: Coccinellidae), an invasive species, has swiftly spread out in Europe. This aposematic and highly polymorphic insect is problematic in several ways. It seems to outcompete native coccinellid species; they can group on fruits in vineyards and spoil wine when crushed with grapes; they can aggregate by hundreds in houses to winter.

This study aims to investigate the relations between the chemical defences of H. axyridis and warning signals expressed by this bug. To achieve this goal, two original quantification methods of H. axyridis secondary metabolites have been undertaken. The first one allows Fast-GC quantifying harmonine, its main defence alkaloid. The other one permits to determine its emission rate of 2-isopropyl-3-methoxypyrazine, a highly odorous compound probably involved in the communication between individuals and in the aposematism of this insect. This last point will be detailed in another poster.

Firstly, a method of extraction, acetylation and purification of harmonine from H. axyridis has been devised to obtain a calibration standard because this molecule is not commercially available. The identity of the extracted compound has been confirmed by mass spectrometry and multidimensional NMR. The GC purity of this N,N-diacetylharmonine is of 100%. During the experiments, its stability in acetone solution at 4°C has been verified. Using this purified molecule, an original method of harmonine quantification has been developed by fast gas chromatography and validated using accuracy profile. The method has been validated for concentrations ranging from 28µg/ml to more than 264µg/ml, with a maximal error risk of 20%. From about 100µg/ml, peaks symmetry decreased, but did not influence the quantification. That accuracy profile validation has been compared with a more classical validation using AOAC (2006) norm, which confirmed the first validation. This method allows quantifying harmonine in one single insect. Harmonine has been quantified in 80 H. axyridis divided into four groups of at least 15 individuals on basis of the morph type and the elytra hue. Males and females harmonine contents did not significantly differ. The mean harmonine content was of 106.6µg/individual with a standard deviation of 80.2µg/individual. The results distribution obtained was not normal and a strong variability between individuals was observed, which explains the high standard deviation.

These results and those of the pyrazine and colour analysis have been statistically analyzed in order to show potential correlations between these factors. It would appear that elytra colour is linked with the insect harmonine content. Individuals with a non-melanic orange colour presented harmonine content significantly higher than other studied colours. The harmonine content does not seem linked to the pyrazine emission rate.

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