Determination of Water-Soluble Components of Abdominal Secretion of Grasshopper (Chorthippus spp.) by GC/MS/MS in Search for Potential Wound Healing Agents.

Wound healing is still a serious medical problem due to process complexity and lack of effective medicaments. This is particularly true in the treatment of wounds arising in the course of such diseases as AIDS or diabetes. Therefore, scientific efforts are focused on the search for new compounds of natural origin, which could be used as medicines or evaluated for subsequent drug design. In folk medicine, grasshopper (Chorthippus spp.) abdominal secretion has been used to accelerate the wound healing process. In this context, the knowledge of the composition of grasshopper abdominal secretion is crucial. The aim of this study was to determine the main water-soluble components of grasshopper abdominal secretion with the use of GC/MS/MS. Liquid-liquid extraction was used as a pretreatment method to clean up, concentrate and fractionate compounds from the complex insect matrix. To obtain more stable and volatile compounds, necessary for GC/MS/MS analysis, a double-step derivatization process was carried out with the use of methoxyamine hydrochloride and a mixture of bis-N,O-trimethylsilyl trifluoroacetamide and chlorotrimethylsilane. As a result, 2,108 compounds were identified, mainly as amino acids, carbohydrates and organic acids. Some of the identified compounds are emphasized due to antimicrobial, antifungal or antioxidant activities reported in the literature. Moreover, a set of compounds characteristic for Chorthippus spp. samples has been selected. In the last part of the study, a statistical analysis was performed to demonstrate differences in composition of aqueous fractions of abdominal secretions from grasshoppers collected at two distant locations: Starogard Gdański and Łubianka meadows.