Spatially resolved chemical analysis of cicada wings using laser-ablation electrospray ionization (LAESI) imaging mass spectrometry (IMS).

Laser-ablation electrospray ionization (LAESI) imaging mass spectrometry (IMS) is an emerging bioanalytical tool for direct imaging and analysis of biological tissues. Performing ionization in an ambient environment, this technique requires little sample preparation and no additional matrix, and can be performed on natural, uneven surfaces. When combined with optical microscopy, the investigation of biological samples by LAESI allows for spatially resolved compositional analysis. We demonstrate here the applicability of LAESI-IMS for the chemical analysis of thin, desiccated biological samples, specifically Neotibicen pruinosus cicada wings. Positive-ion LAESI-IMS accurate ion-map data was acquired from several wing cells and superimposed onto optical images allowing for compositional comparisons across areas of the wing. Various putative chemical identifications were made indicating the presence of hydrocarbons, lipids/esters, amines/amides, and sulfonated/phosphorylated compounds. With the spatial resolution capability, surprising chemical distribution patterns were observed across the cicada wing, which may assist in correlating trends in surface properties with chemical distribution. Observed ions were either (1) equally dispersed across the wing, (2) more concentrated closer to the body of the insect (proximal end), or (3) more concentrated toward the tip of the wing (distal end). These findings demonstrate LAESI-IMS as a tool for the acquisition of spatially resolved chemical information from fragile, dried insect wings. This LAESI-IMS technique has important implications for the study of functional biomaterials, where understanding the correlation between chemical composition, physical structure, and biological function is critical. Graphical abstract Positive-ion laser-ablation electrospray ionization mass spectrometry coupled with optical imaging provides a powerful tool for the spatially resolved chemical analysis of cicada wings.

New Method of Analysis of Lipids in Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) Insects by Direct Immersion Solid-Phase Microextraction (DI-SPME) Coupled with GC-MS.

Lipids play an essential role in providing energy and other physiological functions for insects. Therefore, it is important to determine the composition of insect lipids from cuticular and internal tissues for a better understanding of insect biology and physiology. A novel non-derivatization method for the analysis of lipids including fatty acids, hydrocarbon waxes, sterols in Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) was explored using the direct immersion solid-phase microextraction (DI-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). Nine extraction solvents, acetonitrile, methanol, hexane, ethanol, chloroform, acetonitrile and ethanol (1:1 v/v), acetonitrile and water (1:1 v/v), ethanol and water (1:1 v/v) and acetonitrile and ethanol and water (2:2:1 v/v/v) were selected and evaluated for the extraction of insect lipids with DI-SPME fiber. Acetonitrile extraction offered the best qualitative, quantitative, and number of lipids extracted from insects samples results. Acetonitrile extracted high-boiling point compounds from both species of tested insects. The range of hydrocarbons was C25 (pentacosane) to C32 (dotriacontane) for T. castaneum and C26 (11-methylpentacosane) to C34 (tetratriacontane) for R. dominica. The major compounds extracted from the cuticular surface of T. castaneum were 11-methylheptacosane (20.71%) and 3-methylheptacosane (12.37%), and from R. dominica were 10-methyldotriacontane (14.0%), and 15-methyltritriacontane (9.93%). The limit of detection (LOD) for the n-alkane compounds ranged between 0.08 (nonacosane) and 0.26 (dotriacontane) µg/g and for the fatty acids between 0.65 (arachidic acid) to 0.89 (oleic acid) µg/g. The study indicated that DI-SPME GC-MS is a highly efficient extraction and a sensitive analytical method for the determination of non-derivatized insect lipids in cuticular and homogenized body tissues.