Network Pharmacology‐Based Screening and HPLC Quantification of Antioxidant Compounds in Sojae Semen Praeparatum

Sojae Semen Praeparatum (SSP), commonly known as fermented soybean, is a traditional Chinese medicinal product derived from Glycine max (L.) Merr., renowned for its efficacy in relieving exterior syndromes. To investigate the material basis underlying the ability of SSP to relieve exterior syndromes, network pharmacology was employed as a preliminary screening approach. Genistein and daidzein were identified as the principal bioactive isoflavones in SSP, suggesting that antioxidant mechanisms may contribute to its classical efficacy in dispelling exterior pathogens. Guided by the network pharmacology results, a reliable HPLC method was developed and validated on a Shimadzu LC-16 system. The method employed gradient elution with a mobile phase consisting of acetonitrile and acetic acid/water, with analytes detected at 260 nm. Under the optimized conditions, the validated HPLC method demonstrated excellent performance, with outstanding linearity, precision, repeatability, stability, and recovery. Quantitative analysis using the validated HPLC method confirmed that laboratory-fermented SSP samples consistently met and exceeded the Chinese Pharmacopoeia requirement. These findings provide a scientifically grounded quality control method for SSP and support its consistent use in clinical applications while highlighting the need for further experimental studies to confirm the proposed antioxidant mechanisms.