Abstract: High-performance liquid chromatography (HPLC) experiment is an important instrumental analysis experiment for chemistry majors. However, conventional HPLC experiments designed for instructional purposes often employ oversimplified analytes, making them lack practicality, cutting-edge relevance, and engagement. Crucially, the quantitative chromatographic analysis in traditional teaching laboratories relies on physical/chemical separation, exhibiting suboptimal efficiency and universality while failing to incorporate digital technologies in higher education. To address these limitations, we developed an innovative chromatographic experiment integrating digital technology: the determination of benzenediol isomers in cosmetics through mathematical separation-enhanced HPLC. Conventional HPLC methods for detecting illegally added hydroquinone, catechol, and resorcinol in cosmetic products face challenges including peak overlap, unknown interferences, and poor method transferability. Our approach employs chemometric mathematical separation techniques to resolve HPLC data, eliminating the need for extensive sample pretreatment. This methodology successfully obtains fully selective signals for each component despite peak coelution and unknown matrix effects, enabling precise quantification of three isomers in complex cosmetic matrices. This digitally enhanced experiment fosters students’ innovative thinking and develops their digital literacy. The implementation of “mathematical separation” establishes a robust “3S+3A” analytical approach that not only enhances students’ comprehensive problem-solving abilities but also provides innovative ideas for establishing standard chromatographic detection methods. Rooted in real cosmetic analysis scenarios, it fosters students’ practical skills and professional competence while thoroughly implementing the concept of green analytical chemistry.

Key words: Benzenediol, High-performance liquid chromatography, Mathematical separation, Alternating trilinear decomposition algorithm