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Development of a sensitive and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC–MS/MS) method.Application of UFLC–MS/MS for the determination of RA, sal D, LA and sal B in SAFI in rat plasma.The first pharmacokinetic study of salvianolic acid for injection (SAFI).Investigation of dose-dependent pharmacokinetics of SAFI.A simple, sensitive and selective ultra-fast liquid chromatography-tandem mass spectrometry (UFLC–MS/MS) method was established for simultaneous determination and pharmacokinetic study of rosmarinic acid (RA), salvianolic acid D (Sal D), lithospermic acid (LA) and salvianolic acid B (Sal B) in rat plasma after intravenous administration of salvianolic acid for injection (SAFI). Three doses of administration, containing 14, 28 and 56 mg/kg, were investigated in this study. Plasma samples were pretreated using protein precipitation (PP) with pre-cooled acetonitrile. Chromatographic separation was achieved on a CORTECS™ UPLC C18 column (1.6 μm, 2.1 × 100 mm) with a mobile phase composed of 0.1% formic acid aqueous (V/V) and 0.1% formic acid acetonitrile (V/V). Analytes were detected using electrospray ionization (ESI) source in negative ionization mode and quantified in multiple reaction monitoring (MRM) mode. The validated method is stable and reliable. No significant difference of half lives (t1/2) of four analytes at three doses was observed. Area under the curve (AUC0-∞) and peak concentration (Cmax) of the four analytes demonstrated a linear increase in across the doses with the linear correlation r of each analyte at three doses were greater than 0.95. It indicated that the pharmacokinetic behavior of SAFI is positively related to dose at the range of 14–56 mg/kg.