ABSTRACT
Objective To establish a HPLC method for simultaneous determination of seven active components in Fufang Danshen tablets and Fufang Danshen dripping pills. Methods These seven compounds were analyzed simultaneously with a Zorbax C18 column by gradient elution using acetonitrile-0. 1% phosphoric acid solution as mobile phase, the flow rate was 1 mL·min-1 and the detection wavelength was set at 203, 270 and 281 nm, respectively. Results All the seven components showed good linear relation between peak area and concentration of the test, and the average recoveries were between 95. 1%-100. 4%. Tanshinone ⅡA was not detected in samples of dropping pills. Conclusion The HPLC method to determine the components including tanshinone ⅡA, salvianolic acid B, propanoid acid, protocatechuic aldehyde, notoginsenoside R1 , ginsenoside Rg1 and ginsenoside Rb1 of the two different Danshen preparations has been established, and it has the advantages of simplicity, high precision, good repeatability, and can be used for the quality control of two kinds of Fufang Danshen preparations. The content of tanshinone Ⅱ A in Fufang Danshen tablet was distinctly higher than that of dropping pills.
ABSTRACT
This study was aimed to establish the HPLC fingerprint of liposoluble components inDanshen capsule for quality evaluation. HPLC was run on a Welch ultimate XB-C18 column (250 mm × 4.6 mm, 5 μm) with acetonitrile-water as mobile phase in gradient elution at a flow rate of 1 mL·min-1. The column temperature was 25℃. The detection wavelength was 270 nm. The injection volume was 10μL. The results showed that 7 and 11 common peaks were selected as the HPLC fingerprint of liposoluble components inDanshen capsule from two manufactures, respectively. The similarities of 20 batches of samples were above 0.9. It was concluded that the analysis was stable and reproducible, which can be used as a basis for evaluating the quality control of liposoluble components in Danshen capsule.
ABSTRACT
It is well known that puerarin possesses protective activity on neurodegenerative diseases. However, the exact path way involved in the protective effect of puerarin on MPP+ -induced cell death is unclear. In this study, we focused on mitochondria im pairment in the apoptotic process of MPP+ -elicited SH-SY5Y cells and detected the protection of puerarin. As evidenced by Trypan blue assay, the cell viability was significantly decreased by 1 mmol x L(-1) MPP+, but reversed by different concentrations puerarin pre treatment. Flow cytometer analysis revealed that MPP+ -induced SH-SY5Y cells apoptosis and arrested the cells in G2/M phase, where as puerarin pretreatment concentration dependently reversed the apoptosis ratio. In addition to the apoptosis ratio, 50.0 micromol x L(-1) puerarin pretreatment even altered the MPP+ -induced G2/M phase arrest. JC-1 assay suggested that MPP+ significantly opened MMP of the SH-SYSY cells; pretreatment with puerarin attenuated the deterioration of the MMP. Both ELISA and Western blotting showed that puerarin prevented the release of cytochrome c from the mitochondrial interior to the cystol elicited by MPP+. DNA ladder showed that typical DNA ladder was present in the MPP+ -induced SH-SY5Y cells. Additionally, MPP+ enhanced caspase-9 and caspase-3 ac tivity, respectively, while not caspase-8. However,the enhancement was concentration dependently blocked by puerarin pretreatment. Taken together, puerarin can modulate mitochondrial membrane potential and inhibit the cytochrome c releasing-caspase cascade to pre vent MPP+ -induced cell injury.