Simultaneous Determination of Hydrophilic and Lipophilic Compounds in Radix Salviae Miltiorrhizae by HPLC

Objective To establish a method for determination of the hydrophilic of danshensu,protocatechuic aldehyde,salvianolic acid B and lipophilic compounds of dihydro-tanshinone I,cryptotanshinone,tanshinone I and tanshinone IIA in Radix Salviae Miltiorrhizae. Methods Chromatography was performed on a Lichrospher-C18 column(250 mm ×4.6 mm i.d.;5 μm particle size).The separation was achieved by a linear gradient elution.Mobile phase was the mixture of solvent A(1% aqueous formic acid) and solvent B(acetonitrile).The gradient elution was conducted as follows: 10%-20% B in 0-10 min,20%-33% B in 10-27 min,33%-70% B in 27-30 min,and 70%-85% B in 30-50 min.The injection volume of 10 μL and the UV wavelength of 280 nm were used in the analysis.The flow rate was 1.0 mL·min-1 and the column operated at 20 ℃. Results A good linearity was obtained with the correlation coefficients of tanshinol,protocatechualdehyde,salvianolic acid B,dihydro-tanshinone I,cryptotanshinone,tanshinone I and tanshinone IIA ranged from 0.037 2-118.000 0 mg·L-1,0.017 4-8.900 0 mg·L-1,0.141-450.000 mg·L-1,0.018 8-56.800 0 mg·L-1,0.049 7-158.000 0 mg·L-1,0.021 5-78.500 0 mg·L-1,0.096 2-308.000 0 mg·L-1(R2≥0.999 2),respectively.The average recovery rates of the four investigated compounds were in the range of 98.5%-101.5% for all with RSD below 1.98%(n=6).The intra-day and inter-day precision of the assay were no more than 2.16%(n=6). Conclusion The method is accurate and reliable with good reproducibility,and can be used as a determination standard for Radix Salviae Miltiorrhizae.