Simultaneous determination of cryptotanshinone,tanshinone I and tanshinone II_A in ethanol extract of Salviae Miltiorrhizae Radix et Rhizoma by ultra-performance liquid chromatography

Objective: To establish an ultra-performance liquid chromatography method for simultaneous determination of three representative tanshinones( cryptotanshinone,tanshinone Ⅰ and tanshinone ⅡA) in ethanol extract of Salviae Miltiorrhizae Radix et Rhizoma. Methods: The method was performed on a Waters Acquity UPLC BEH Shield RP18( 1. 7 μm,2. 1 mm × 100 mm) analytical column employed with a guard column of Phenomenex SecurityGuard C18Cartridges Polar-RP( 4 mm × 3. 0 mm) at the column temperature of 35 ℃. The gradient mobile phase consisted of acetonitrile( A)-5% acetonitrile aqueous solution containing 0. 1% formic acid( pH 2. 5)( B) with a flow rate of 0. 25 mL·min-1( 0-5 min,52%A→70%A; 5-7 min,70%A→80%A; 7-10 min,80% A→ 52% A; 10-12 min,52% A). The injection volume was 5 μL. The UV detection wavelength was set at 269 nm. Results: The relationship between the concentration and the peak area of cryptotanshinone,tanshinone Ⅰ and tanshinone ⅡAwas all in good linearity( r 0. 999) over the linear ranges. The average recoveries( n = 6) of the investigated compounds were 96. 7%( RSD = 1. 3%),96. 4%( RSD = 0. 94%) and 98. 5%( RSD = 1. 2%); the chromatographic peaks of investigated compounds and coexisting components were well isolated. Conclusion: Compared with routine liquid chromatography,this method with high separation efficiency and good repeatability can be used as a reference for improvement of laboratory methods and make preparation for improvement of the quality standard of tanshinone extract.