## 1 Introduction  
Lipids are the main biological compounds in animals and plants [1], including fatty acids and hydrocarbon waves on and in insect bodies [2]. Fatty acids are the most basic form of biological lipidsand are usually bound with other compounds to build more composite lipids, such as triglycerides, which are energy stores [3]. The cuticular lipid layer of the insect consists of different chemicals such as a long chain of hydrocarbons and fatty acids [4]. An analysis of the cuticular lipids of _Acanthoseclides obtetcus_ showed that adults contain a variety of chemicals such as hydrocarbons, triacylglycerglycerols, fatty acid esters, free fatty acids, sterols, aldehydes, ketones and alcohols [5]. Hydrocarbons are the major lipid category in the cuticle of insects, including straight-chain saturated and unsaturated hydrocarbons [6], which in some species reach to more than 90% of cuticular lipids and are usually a mixture of components including n-alkanes, branched methyl-alkanes and ethyl-alkanes [2]. An analysis of cuticular lipids from adults of _Zygogranma exclamationis_ showed that large amounts of lipids in the cuticle of males and females were hydrocarbons ranging from C23 (tricosene) to C56 (trimethyltripentacontane) [7]. The functions of insect cuticular hydrocarbons have evolved to not only keep water in the organism but to also play an essential role in communication among the population individuals and between individuals of different sexes [2; 8; 9]. A study on the adults of _Drosophila melanogaster_ demonstrated that increased desiccation resistance is linked with the total amount of cuticular hydrocarbons [10]. Cuticular lipids have also been reported to protect insects from harmful pathogens [2; 11]. The composition of cuticular lipids varies among the insect species, and it is a reflection of the genetic structure and changes induced by ecological circumstances [12]. Therefore, an analysis of insect lipids is fundamental to understand insect metabolism and physiology. The extraction of lipids from insects is the first critical step that leads to a reliable qualitative and quantitative analysis of insect lipids [1]. Numerous methods have been used to extract insect lipids; however, extraction solvents and methods are the core of a reliable extraction process that transfers lipids from a matrix to the liquid phase [1; 13; 14; 15], thus enabling coupling with various analytical methods [1]. Examples include using a single solvent like hexane to extract cuticular hydrocarbons from six species of flies and an analysis with gas chromatography-mass spectrometry (GC-MS) [13], extraction with chloroform and an analysis with high-performance thin-layer chromatography (HPTLC) to determine surface lipids of _Bemisia argentifolii_[14], and extraction with dichloromethane and an analysis with gas chromatography-Flame ionization detector (GC-FID) for profiling the cuticular hydrocarbons of four _Periplaneta_ species including _Periplaneta brunnea_, _Periplaneta filginosa_, _Periplaneta australisiae_ and _Periplaneta americana_[15]. Commonly, the use of two solvents, such as hexane/chloroform, have been used for extraction of the cuticular lipids from the _Osmia lignaria_ and _Megachile rotundata_ bees and the _Aleurodicus dugesii_ giant whitelfly, which were then analyzed with GC-FID and GC-MS [16]. Solid-phase microextraction (SPME) has been used to extract insect lipids, especially cuticular lipids [1]. This extraction is done either by rubbing the SPME fiber onto the surface of the insect cuticle [17; 18] or by using the headspace solid-phase microextraction (HS-SPME) method [19]. However, these procedures are not suitable for the analysis of semi-volatile compounds. Keeping this in mind, a novel method of direct immersion solid-phase microextraction (DI-SPME) in different solvents coupled with GC-MS has been used.