Seminal plasma chemical profiles provide distinct and metabolic phenotypes suited for fertility study monitoring. Profiling via GC-MS is favourable as it provides robust measures across many core primary metabolic pathways; including amino acids, TCA metabolites, carbohydrates and some lipids (e.g. steroids & fatty acid). A protocol for simple preparation of seminal plasma samples intended to undergo GC-MS analysis is described and enables large-scale semi-quantitative profiling.