Comprehensive Profiling of Chemical Constituents in Niaoduqing Granules by Ultra‐High‐Performance Liquid Chromatography‐Q‐Exactive Orbitrap Mass Spectrometry and Identification of Potential Renoprotective Active Components

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Niaoduqing Granules (NDQG) is a Chinese patent medicine preparation composed of more than 10 herbal ingredients and is widely used in the clinical treatment of chronic kidney disease. Nevertheless, comprehensive and systematic investigations into the chemical constituents and pharmacodynamic material basis of NDQG remain insufficient. Herein, a binary chromatographic fingerprinting strategy was developed for the systematic characterization of the chemical constituents in NDQG using both acidic and alkaline mobile phase systems. Independent collection and combined analysis of acidic/alkaline mobile phases enabled the effective separation of compounds with significant polarity differences and varying chromatographic properties in NDQG, resulting in good peak shapes. Using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry, this study identified 187 chemical components from the NDQG preparation (58 compounds were qualitatively confirmed by reference standards). The prototype components distributed to plasma and kidney tissues in C57BL/6 mice following oral administration of NDQG were further investigated. 40 components were detected in the blood circulation, and 36 were identified in kidney tissues. Using a cobalt chloride-induced Human Kidney-2 (HK-2) cell injury model, further activity screening of 28 candidate compounds revealed that nine compounds (formononetin [FMN], ononin [ONO], liquiritin, ferulic acid, quercetin, rosmarinic acid [ROA], paeoniflorin, calycosin [CS], and salvianolic acid B [SAB]) effectively alleviated HK-2 cell injury. Among them, FMN, ONO, ROA, CS, and SAB significantly inhibited the mRNA expression of both tumor necrosis factor-α and interleukin-18, thereby exerting favorable anti-inflammatory effects. These components may act as important lead compounds, providing valuable implications for drug discovery and development. In summary, combining component characterization and activity screening, this study not only provides key data for the quality control of NDQG but also furnishes a scientific basis and novel insights for clarifying its pharmacodynamic material basis and supporting the subsequent development of innovative drugs.