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Aim: To develop a method for analysis of antitumor annonaceous acetogenins in Annonaceae plants by HPLC., Methods: Squamostatin-B (1), squamocin (2) and annonin-VI (3) were used as standard substances. Chromatography column was a Rp-18; the mobile phase was methanol-water (90:10); the flow rate was 1.0 mL.min-1 and the detecting wavelength was 220 nm., Results: A linear range was obtained from 2.3 to 13.8 micrograms with a good correlation. The recoveries of (1), (2) and (3) were 100.3%, 100.3% and 100.0%, respectively., Conclusion: This method was developed for the analysis of acetogenins by HPLC for the first time. The method is rapid, accurate and suitable for the analysis of the antitumor acetogenins in Annonaceae plants.

Aim: To investigate the mechanism of apoptosis of HL60 cells induced by the annonaceous acetogenin, squamocin., Methods: Induction of apoptosis was determined through Hoechst33258 dye staining and DNA agarose gel electrophoresis. Expression of the proteins was detected using Western blot analysis. Caspase-3 activity was detected using caspase-3 kit., Results: Treatment of HL-60 cells with squamocin resulted in extensive nuclear condensation, DNA fragmentation, cleavage of the death substrate poly(ADP-ribose) polymerase (PARP) and induction of caspase-3 activity. Pretreatment of HL-60 cells with caspase-3 specific inhibitor DEVD-CHO prevented squamocin-induced DNA fragmentation, PARP cleavage and cell death. Stress-activated protein kinase (SAPK/JNK) was activated after treatment with squamocin in HL-60 cells., Conclusion: These results suggest that apoptosis of HL-60 cells induced by squamocin require caspase-3 activation, and could be related to SAPK activation.