The application of chemical analysis on bituminous materials has increased drastically over the past decades. One of the most common spectroscopic methods used in the field of research is Attenuated Total Reflection (ATR) Fourier Transform Infrared (FTIR) spectroscopy. Since ATR-FTIR is a surface sensitive method, sample or specimen handling of a complex material like bitumen prior to its analysis needs to be considered, especially for people new to the field or analysis technique. This study looks at the impact of heating time and temperature as well as storage time and conditions on the oxidation of the bituminous specimen. Four binders from the same crude oil source but different specification classes (unmodified and styrene–butadiene–styrene polymer modified) and two binders from different crude oil sources were investigated. The results show that heating small quantities of bitumen at 180 °C for up to 30 min has little impact on the formation of oxidized species, when proper thermal monitoring is conducted. Special cases where oxidation does occur are reported in detail. Furthermore, strong oxidation is induced by day light, when bitumen is stored behind glass with no UV radiation present, which can reach short-term ageing level within 1 h. Thus, heating bitumen at 180 °C for 5–10 min followed by storage in the dark, climatized room and measured within one hour after preparation is recommended. These results should act as recommendation for future specimen handling prior to FTIR spectroscopic analysis to ensure unbiased and comparable measurements.
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