Study On Optimal Process Of Same Peak Value Of CPT And Hcpt Of Seed Bud In Camptotheca Acuminata

Camptothecin (CPT) and Hydroxycamptothecin (HCPT) in Camptotheca acuminata possess good bioactivity. Being its stronger bioactivity of anti-cancer and less side effect, HCPT becomes another new anti-cancer medication in international market following taxol. But natural content of HCPT in Camptotheca acuminata seeds is about 1/1000000, it must be obtained by synthesizing. The method is high cost, long process and low yield. Previous research discovered the theory of accumulation by biotransformation during seeds germination in Camptotheca acuminata can be applied in mass-produce HCPT.Deeply research has been done on seed germinative condition of Camptotheca acuminata. Environmental influence factors, such as temperature , light and water et al were comparatively analyzed, Contents of CPT and HCPT during different germinative period were monitored dynamically by changed wave length HPLC, the same peak value time of CPT and HCPT was forecasted. For improving contents of CPT and HCPT in germinative seeds at the same peak value time, the method and the mechanism of contents CPT and HCPT in Camptotheca acuminata were discussed and analyzed.1 .Established sample preparation method by ultrasonic extraction: fresh bud were crushed and extracted by 70 percent of ethanol (extraction solvent) at 40 and 15kHz for 60 minutes.2.The HPLC analysis method of CPT and HCPT at the same time were determined: Reverse pillar (25cm 4.6mm, 5m); the mobile phase is Acetonitrile : water (3:7); the flow rate was maintained at 1.0mL/min; measure time was 20 minutes with the wavelength at 266nm from 8 to 10 minutes,254nm from 8 to 20 minutes; temperature of pillar was 25 .3.The optimum germinative process were determined of Camptotheca acuminata seed. It is that the seeds were disposed at 5 for 30 days before germination; at ultrasonic condition at 25 for 10 minutes,20kHz;soaked in cold water for 48 hours; germinated at 30 day/ 20 night(light).4.From 11 to 13 days after seeds sucking, that is the time when the cotyledon came out, the content of HCPT in the bud is the highest.This process using enzyme system in natural seed to control environmental factors which influence seed secondary metabolism, the content of HCPT in bud were increased. Germination was terminated when the content of HCPT is highest. The buds can provided the optimum extract materials possessing high content CPT and HCPT, and lay the base of theory and practice to use seed and develop the resources of Camptotheca acuminata with high efficiency and profits.