Cuticular Hydrocarbon Profiling by Fractionation and GC-MS in Socially Parasitic Ants

Hironori Iwai; Nobuaki Kono

doi:10.21769/BioProtoc.4772

Cuticular Hydrocarbon Profiling by Fractionation and GC-MS in Socially Parasitic Ants

Bio Protoc. 2023 Jul 5;13(13):e4772. doi: 10.21769/BioProtoc.4772.

Authors

Hironori Iwai¹, Nobuaki Kono^{1

2

3}

Affiliations

¹ Institute for Advanced Biosciences, Keio University, Tsuruoka, Japan.
² Graduate School of Media and Governance, Keio University, Tsuruoka, Japan.
³ Faculty of Environment and Information Studies, Keio University, Fujisawa, Japan.

Abstract

Ants use cuticular hydrocarbon (CHC) as a semiochemical for recognizing their nestmates. For socially parasitic ants, deceiving the CHC is an important survival strategy. Profiling and quantifying CHC is a potent approach to understanding such nestmate discrimination behavior. Thus, a highly efficient, stable, and reproducible extraction method for CHC is essential for this purpose. This paper describes a method for socially parasitic ants to disguise the host species' CHC profile under laboratory conditions, as well as the extraction and measurement of CHC from ants (from a previous study). First, the artificial isotopic substance is applied to the host worker; then, the socially parasitic ant disguises the host-like CHC profile against the above host worker. Next, the CHC is extracted and fractionated from a socially parasitic ant using hexane and silica gel. After concentrating the fractionated product, this product is then used for measurement by gas chromatographymass spectrometry (GC-MS). The CHC extraction protocol described in this paper may be used for various ant species.

Keywords: Ant; Chemical disguise; Cuticular hydrocarbon (CHC); Gas chromatographymass spectrometry (GCMS); Myrmecology; Semiochemical; Social insect; Social parasite.