Development of a GC-Based Method for Hedysari Radix Polysaccharide Quantification in Rats and Its Application to Pharmacokinetic and Tissue Distribution Studies

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This study aimed to establish an accurate method for quantifying Hedysari Radix polysaccharide (HPS) components (HPS-50 and HPS-80) in rat biological samples and to apply it for comparative pharmacokinetic and tissue distribution studies. An integrated gas chromatography (GC)-based method was developed for this purpose. The sample pretreatment protocol involved hot water precipitation, followed by lyophilization. The dried powdered samples were subsequently hydrolyzed using trifluoroacetic acid, followed by oximation and acetylation derivatization, and the final detection was performed using GC-FID. Arabinose, an endogenously absent monosaccharide employed as a surrogate biomarker due to its natural absence in rats. The method was systematically validated for linearity, precision, accuracy and stability. The validation results confirmed that the method was linear (R² > 0.99), precise (RSD < 15%), accurate (85–115%), and stable. Pharmacokinetic analysis revealed that compared to HPS-50, HPS-80 exhibited a significantly prolonged half-life (9.32 h vs. 2.49 h), higher systemic exposure (AUC₀–∞: 4.81 vs. 1.71 h·mg/mL), and slower clearance rate (20.8 vs. 58.37 mL/h/kg). Tissue distribution studies showed that HPS-80 had higher uptake and more prolonged retention in metabolic organs (liver and kidneys) and immune organs (spleen). The established GC-based method is accurate and reliable, providing a robust tool for in vivo studies of HPS and potentially facilitating the rationalization of clinical medicine and individualized administration of HPS.

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