Comprehensive quality standardization of Haizhixiaolin capsules via multi-component fingerprinting and metabolomic

Haizhixiaolin Capsule (HZXL), a traditional Chinese medicine hospital preparation for treating chronic prostatitis and urinary disorders, consists of ten herbal components. Due to the complexity of its multi-component nature, establishing a robust quality control system remains challenging. This study presents the first systematic quality evaluation of HZXL using high-performance liquid chromatography (HPLC) fingerprinting combined with ultra-high-performance liquid chromatography-Orbitrap mass spectrometry (UHPLC-Orbitrap MS) untargeted metabolomics. Based on high-performance liquid chromatography, four reference standards (caffeic acid, geniposide, myricitrin, and calceolarioside B) were used as markers for the fingerprint analysis of ten batches of HZXL, and the results demonstrated excellent similarity (>0.930). Using liquid chromatography-mass spectrometry (LC-MS), untargeted metabolomics identified 152 compounds, predominantly flavonoids, iridoid glycosides, phenolic acids, and organic acids. Chemometric analysis, including hierarchical cluster analysis (HCA) and principal component analysis (PCA) on six representative batches, revealed high overall batch consistency with only minor inter-batch variations. The Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) combined with entropy weighting was applied to screen core quality markers from the 152 identified compounds. Chlorogenic acid, glycyrrhetinic acid, geniposide, luteolin, quercetin, and caffeic acid ranked highest based on PC loadings, stability, relative abundance, and pharmacological activity scores. This multi-component quality evaluation framework, integrating fingerprinting, high-resolution metabolomics, and advanced chemometrics, provides a scientific basis for establishing comprehensive multi-indicator quality standards, improving batch-to-batch reproducibility, and supporting the modernization and internationalization of HZXL.