Minutes to measure: Ultra-fast and green HPLC-MS for flavonoid profiling in Citri Reticulatae Pericarpium

Citri Reticulatae Pericarpium (Chachi and Common mandarin), a dual-purpose traditional Chinese medicine (TCM) and food, contains bioactive flavonoids such as hesperidin, nobiletin, and tangeretin, which exhibit antioxidant, anti-inflammatory, and cardioprotective properties. However, conventional high-performance liquid chromatography with ultraviolet detection methods for their quantification are time-consuming (>80 min), environmentally unsustainable (high organic solvent use), and lack sensitivity for low-abundance components (e.g., nobiletin, tangeretin). To overcome these limitations, this study developed a rapid, eco-friendly method by integrating vortex-assisted matrix solid-phase dispersion with Poroshell column and high-performance liquid chromatography-mass spectrometry (HPLC-MS), reducing analysis time to 4 min (1 min extraction + 3 min HPLC-MS) and cutting solvent consumption by 75%. The optimized protocol demonstrated highly sensitive and efficient enabling precise quantification of target flavonoids in 24 batches Citri Reticulatae Pericarpium (12 batches each of Chachi and Common mandarin). The contents of hesperidin, nobiletin, and tangeretin in 24 batches of samples were 28.18–78.56, 0.24–3.30, and 0.11–2.80 mg g −1 dry weight, respectively. Finally, the greenness of the developed method was evaluated using the green analytical procedure index, which indicated a superior environmental profile with five green regions. Additionally, the blue applicability grade index tool was employed, resulting in a score of 80, reflecting high applicability and environmental friendliness. Results validated the method's robustness for rapid quality assessment of Citri Reticulatae Pericarpium, supporting TCM standardization and clinical safety. This advancement aligns with green analytical chemistry principles and offers a transformative approach for ultra-fast, sustainable herbal analysis.