Yw. Guo et al., IMMUNOLOGICAL DETECTION AND QUANTITATION OF 10-DEACETYLBACCATIN-III IN TAXUS SP PLANT AND TISSUE-CULTURES, Biological chemistry Hoppe-Seyler, 375(4), 1994, pp. 281-287
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.