Fine quantitation of novel trace taxans in suspension-cultured Corylus avellana L. cells by photo diode array HPLC technique

Taxanes are widely known as great family of antitumor compounds. Identification of certain taxanes, particularly taxol, has opened new perspectives for further researches in plant and medical sciences. The aim of the present study was to manipulate chromatographic method in order to detect and quantified novel trace taxanes in suspension-cultured hazel (Corylus avellana L.) cells. A rapid growing cell line established from hazel seeds were grown in a modified LS media. The cells were harvested and extracted at days 10 and 15 when based on growth curve they were in the second half of logarithmic growth phase. Separation of taxanes was conducted by a coupled gradient-isocratic high performance liquid chromatography, equipped with a photo diode array detector. Base on chromatographic behavior and UV spectrum, the method enabled us to determine and quantified not only previously reported taxoides i.e., taxol, 10-deacetylbaccatin III, baccatin III, but also trace novel ones e.g., cephalomannine, 7-epi-taxol, 7-epi-10-deacetyltaxol and 10-deacetyltaxol. Certain taxanes i.e., 10-deacetylbaccatin III, baccatin III, taxol, and 10-deacetyltaxol were the most abundant taxanes detected at day 10, while 7-epitaxol, 7-epi-10-deacetyltaxol, cephalomannine were identified and quantified at day 15. Moreover, total content of taxanes was higher in day 15 than day 10. It should be noted that different taxanes bear different applications, for example taxol is directly used in medicine while 10- deacetylbaccatin III, baccatin III, 10-deacetyltaxol are more considered as valuable precursors in semisynthetic production of other taxanes. Therefore, the results presented here can provide approaches in decision making and time management toward extract maximum amounts of a desired taxan.