Optimization of podophyllotoxin extraction method from Linum album cell cultures.

CONTEXT

Suspension cultures of Linum album Kotschy ex Boiss. (Linaceae) accumulate podophyllotoxin (an anticancer agent) and could therefore serve as an alternative source of this important aryltetralin lactone lignan.

OBJECTIVE

The present work compared four podophyllotoxin extraction methods and optimization of the best one by using single factor experiments.

MATERIALS AND METHODS

Linum album cell cultures were established from in vitro plantlets and subcultured in MS medium with hormones every 7-8 days. Four podophyllotoxin extraction methods were assayed and the best one was optimized by single factor experiments, studying the effect of methanol concentration, extraction time, and sonication time.

RESULTS

Cell cultures accumulated enough podophyllotoxin to be analyzed by HPLC. The methanol/dichloromethane and buffer extraction methods were found to be the best. Methanol alone and hot ethanol were not effective for extracting podophyllotoxin.

DISCUSSION AND CONCLUSION

The optimized method based on methanol/dichloromethane extraction combined with HPLC quantification was able to determine small amounts of podophyllotoxin in Linum album cell cultures, showing that this system could constitute a possible alternative source of podophyllotoxin to Podophyllum (Berberidaceae).