Development of a Quality Evaluation Card for Saffron (Crocus sativus L.) by Integrating Chemical, Colorimetric, and Antithrombotic Bioactivity Analyses

• Combines traditional TCM morphology with modern analytical techniques. • Shows a positive link between saffron's antithrombotic effect and its color/key components. • Develops a quality evaluation card based on the established color-efficacy relationship. • Validates the traditional "quality evaluation by appearance" principle scientifically. Saffron ( Crocus sativus L.), a traditional Chinese medicine with both medicinal and culinary uses, was examined to establish a comprehensive quality evaluation system integrating morphological, biological, and chemical approaches, based on the "Quality evaluation by appearance characterization" theory, to enable rapid and scientific assessment of its antithrombotic efficacy. A zebrafish thrombosis model was employed to evaluate 50 saffron batches based on bioactivity. Of these, 43 batches demonstrated significant antithrombotic effects (10 batches, including S1, S4, and S41–S48, classified as Grade 1, with the remainder classified as Grade 2). Chemical fingerprinting identified key discriminatory components, such as picrocrocin and crocin-I, while colorimetric analysis provided objective quantification of color parameters. Results revealed significant correlations between color features (L*, a*, b*) and antithrombotic activity/chemical composition ( p 45% or ≤ 45%) through color comparison. This study offers a rapid, accurate, and practical strategy for evaluating the quality of medicinal and edible traditional Chinese medicines, advocating a shift from single-component analysis to a holistic, efficacy-focused approach.