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In this study, we report a novel ZnO/polyaniline (PANI) nanocomposite optical gas sensor for the determination of acetic acid at room temperatures. ZnO nanorods, synthesized in powder form were coated by PANI (ZnO/ PANI) by chemical polymerization method. The obtained nanocomposites were deposited on glass substrate and dried overnight at room temperature. Structure and optical properties of ZnO/PANI nanocomposite have been studied by using X-ray diffraction, transmission electron microscopy, scanning electron microscopy, diffuse reflectance and photoluminescence spectroscopy. Tests towards acetic acids were performed in the range of concentrations 1–13 ppm. The adsorption of acetic acid on the sensor's surface resulted in the decrease of ZnO/ PANI photoluminescence. The response and recovery time of the sensor were in the range of 30–50 s and 5 min, respectively. The developed sensors showed sensitivity towards acetic acid in a range of 1–10 ppm with the limit of detection of 1.2 ppm. Specially designed miniaturized sensing system based on integrated sensing layer, light emission diode as excitation source and optical fiber spectrometer was developed for the measurement of the sensor signal. The developed sensing system was applied for the investigation of some real sample assessment including the evaluation of storage conditions of ancient cellulose acetate films, which during the degradation are releasing acetic acid. The obtained results suggest that the developed novel optical ZnO/PANI nanocompsite based sensor shows great potential for acetic acid determination in various samples.