IMMUNOLOGICAL DETECTION AND QUANTITATION OF 10-DEACETYLBACCATIN-III IN TAXUS SP PLANT AND TISSUE-CULTURES

A high-sensitive ELISA method was developed for the detection and semi -quantitative determination of 10-deacetylbaccatin III and its structu rally related compounds in crude extract of Taxus sp. plants and tissu e cultures. The antibodies were raised in rabbits using 7- or 10-succi nyl-10-deacetylbaccatin III-BSA conjugate as immunogen. The working ra nge of the assay was from 0.003 to 1.000 ng (0.09 to 31.33 nM) of 10-d eacetylbaccatin III per assay. The cross-reacting material in crude pl ant extract was examined by chromatographic (silica gel CC, HPLC) and immunoassay methods. Study on the evaluation of cross-reacting mate; r ial in crude Taxus plant extracts showed that at least 80% of the immu nosignal correspond to 10-deacetylbaccatin III in the extract. The ELI SA method was applied to investigate the 10-deacetylbaccatin III equiv alent content in crude extracts of 19 plants species including Taxacea e, Taxodiaceae and Pinaceae species. The 10-deacetylbaccatin Ill-like structure was only detected in Taxus and Torreya sp. The results indic ate that this immunoassay is a useful tool for the rapid screening of species, varieties or individual plants out of a wide population. The distribution of 10-deacetylbaccatin III equivalent content in 9-month old Taxus plantlets cultivated in vitro as well as in callus culture w as investigated.