Quality Evaluation of Toosendan Fructus Based on Content Determination and Pharmacological Activity

OBJECTIVE To compare the contents and color values of two representative components， toosendanin and isotoosendanin， in different batches of toosendan fructus， and to compare the effects of different batches of toosendan fructus extracts on the expression of DR5 and the apoptosis of NSCLC cells induced by TRAIL. METHEODS The contents of toosendanin and isotoosendanin in toosendan fructus were determined by UHPLC-QqQ-MS/MS. The analysis was performed on Poroshell 120 SB C₁₈ (2.1 mm×100 mm，1.8 μm) column， mobile phase was 0.1% formic acid aqueous solution and acetonitrile with flow rate of 0.3 mL/min. The injection volume was 1 μL， and the column temperature was 35 ℃. Meanwhile， spectrophotometer was used to determine the color value of the samples. Flow cytometry analysis was used to detect apoptosis and the expressions of cell death receptor DR5 for TRAIL in non-small cell lung cancer cell line A549. RESULTS The contents of toosendanin and isotoosendanin in 10 batches of samples were 1.024~4.128 mg/g and 0.010~0.045 mg/g， respectively. The average of colour value (L^*，a^*，b^*) was 68.56，5.27 and 25.12. The extracts from 10 batches of toosendan fructus could significantly increase the expression of DR5 and restore the apoptotic sensitivity of A549 cells towards TRAIL. CONCLUSION The content determination and color measurement， combined with pharmacological activity can be successfully applied to accurately evaluate the quality of toosendan fructus.