We report the detection of microbial contamination and growth of cinematographic film utilising headspace solid phase micro-extraction coupled with Gas Chromatography–Mass Spectrometry. Microbial Volatile Organic Compounds (MVOCs) are produced only when the mould is actively growing on the cine film. Over 150 volatile compounds were detected from 16 fungal isolates, with over 40 being common to 2 or more isolates. It was found that 1-octen-3ol was produced from 13 of the isolates analysed, 3-octanone from 10 of the isolates and 3-octanol from 4 isolates. These three key chemical markers are indicative of viable fungal growth on cinematographic film precluding the need for traditional microbiology laboratory culture methods. Such an approach would prevent valuable historical footage from being discarded due to health and safety concerns regarding spore inhalation, and would enable safe handling.