Abstract
There is increasing emphasis on the use of new analytical approaches in subject analysis and classification, particularly in respect to minimal sample preparation. Here, we demonstrate that rapid evaporative ionization mass spectrometry (REIMS), a method that captures metabolite mass spectra after rapid combustive degradation of an intact biological specimen, generates informative mass spectra from several arthropods, and more specifically, is capable of discerning differences between species and sex of several adult Drosophila species. A model including five Drosophila species, built using pattern recognition, achieves high correct classification rates (over 90%) using test datasets and is able to resolve closely related species. The ease of discrimination of male and female specimens also demonstrates that sex-specific differences reside in the REIMS metabolite patterns, whether analysed across all five species or specifically for D. melanogaster. Further, the same approach can correctly discriminate and assign Drosophila species at the larval stage, where these are morphologically highly similar or identical. REIMS offers a novel approach to insect typing and analysis, requiring a few seconds of data acquisition per sample and has considerable potential as a new tool for the field biologist.
Highlights
Insect identification and monitoring are essential to a number of diverse fields and settings, seeking to identify and study insect populations to learn more about their place in ecosystems as well as their impact on the environment and other species [1]
A mixture of wild-trapped arthropod species and five laboratory-raised Drosophila species were used for a proofof-principle study to investigate rapid evaporative ionization mass spectrometry (REIMS) suitability for insect analysis and gauge its potential as an identification device
The mass spectra were imported into the model building software packages; Offline Model Builder (OMB-1.1.28; Waters Research Centre, Hungary) and LiveID (Waters, UK), which allow separation of sample groups based on principal component analysis (PCA) and linear discriminant analysis (LDA)
Summary
Insect identification and monitoring are essential to a number of diverse fields and settings, seeking to identify and study insect populations to learn more about their place in ecosystems as well as their impact on the environment and other species [1]. Long-term biodiversity and environmental impact studies [2,3] tend to observe and log the changes and make-up of insect populations In other circumstances, such as biological control in pest management, maintaining the population of certain species is desirable or even necessary to sustain ecosystem balance [4]. Molecular analytical tools have been developed and applied to aid morphological examination and expand capabilities These include cuticular hydrocarbon analysis [26], immunological [27] or protein-based assays [28] as well as mass spectrometry-based applications such as matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) [29]. Easy-to-use high throughput tools capable of handling a variety of samples in vast amounts are still sought after and could provide much-needed support in the wide array of fields requiring rapid insect identification. A mixture of wild-trapped arthropod species and five laboratory-raised Drosophila species were used for a proofof-principle study to investigate REIMS suitability for insect analysis and gauge its potential as an identification device. 2 Our results demonstrate the techniques ability to distinguish species as well as the sex of specimens using models developed from the uninterpreted mass spectra that are derived from aerosol analysis
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