& Hollow fiber liquid phase microextraction combined with high performance liquid chromatography (HFLPME with HPLC) is a simple, rapid, and sensitive method for the study on the pharmacokinetics of the active ingredients of lignans (i.e., schisandrol A, schisantherin A, schizandrin A, schizandrin B, and schizandrin C) from Schisandra chinensis after oral administration to rats. Several important HFLPME parameters, such as type of hollow fiber and extraction solvent, pH of sample phase, ionic strength, centrifugation speed, extraction time, and sample volume, were carefully investigated and optimized. Under optimal conditions, the enrichment factors for the five lignans in urine and plasma were within the ranges from 42.7 to 99.5 and 42.3 to 189.0, respectively. The linear ranges, limits of detection and quantification, precisions, and the mean recoveries and stabilities were satisfactory during the determination of five lignans in urine and plasma. The pharmacokinetics of five lignans was simultaneously studied in rats following intragastric administration of S. chinensis; the pharmacokinetic parameters were also estimated. The validated HFLPME with HPLC is suitable for the preconcentration and determination of trace amount of lignans in biological fluid. The method can also be used in research on the pharmacokinetics of active constituents in traditional Chinese medicines.