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In this research study, a rapid, sensitive, and specific high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry (HPLC–ESI–MS/MS) method was established and validated, in regard to the simultaneous quantification of five sedative and hypnotic lignans (schisandrin, schisandrol B, schisantherin A, deoxyschisandrin, and schisandrin B) in various tissues of rats (including heart, liver, spleen, lung, and kidney). The purpose of the study was to clarify the tissue distribution of the total lignans extract of Schisandra chinensis (SC). Then, the analytes were separated on a MERCK Purospher STAR LP C18 column (250 mm × 4.6 mm, 5 μm), with a mobile phase consisting of 0.05% (v/v) formic acid acetonitrile, and 0.05% (v/v) formic acid water, and a flow rate of 1 mL/min. All of the calibration curves of the five components showed good linearity (r > 0.9950), with ranges of 4.8 to 1920 ng/mL for analytes. The intra-day and inter-day precisions (relative standard deviation [RSD] %) were within 13.76% for all of the analytes. The average recoveries of the five analytes were greater than 85.23%, and the mean value of the matrix effect ranged from 82.3% to 93.4%. The five analytes were confirmed to be stable during the storage, preparation, and analytic procedures. The major target tissues of the total lignans extract of the SC in the rats were the livers and kidneys. [ABSTRACT FROM AUTHOR]