Poulev A, Deus-Neumann B, Bombardelli E, Zenk M H
Lehrstuhl für Pharmazeutische Biologie, Universität München, Federal Republic of Germany.
Planta Med. 1994 Feb;60(1):77-83. doi: 10.1055/s-2006-959413.
A radioimmunoassay as well as an enzyme immunoassay for the quantitation of fmol amounts of the alkaloid colchicine have been developed. The antiserum used for both assays was raised against a conjugate of colchicoside-bovine serum albumin. The crude serum was satisfactory for the performance of the radioimmunoassay. For the enzyme immunoassay, the antibodies had to be isolated and purified by Rivanol treatment with subsequent (NH4)2SO4 precipitation. The measuring range extends from 0.1 to 100 ng colchicine for the radioimmunoassay and from 0.05 to 350 ng for the enzyme immunoassay with detection limits of 125 fmol and 25 fmol, respectively. Both immunoassays cross reacted with colchicoside and 3-demethyl-colchicine up to 80%. The colchicine content in the newly established suspension culture of Colchicum variegatum as well as the influence of various culture media on the colchicine production of this cell culture were investigated with the radioimmunoassay. The enzyme immunoassay was well suited for the quantitation of colchicine in HPLC fractions of Gloriosa and Colchicum seed extracts allowing the rapid, sensitive, and precise determination of the substance under investigation. The preliminary experiments indicate that both colchicine immunoassays can be a useful tool for the analysis of colchicine in tissue and cell culture studies, for analysis of plant extracts as well as for biosynthetic investigations.
已开发出一种用于定量测定飞摩尔量生物碱秋水仙碱的放射免疫分析法和一种酶免疫分析法。两种分析方法所用的抗血清均是针对秋水仙苷 - 牛血清白蛋白偶联物产生的。粗血清用于放射免疫分析表现令人满意。对于酶免疫分析，抗体必须通过利凡诺处理并随后用硫酸铵沉淀进行分离和纯化。放射免疫分析的测量范围为0.1至100 ng秋水仙碱，酶免疫分析的测量范围为0.05至350 ng，检测限分别为125飞摩尔和25飞摩尔。两种免疫分析方法与秋水仙苷和3 - 去甲基秋水仙碱的交叉反应高达80%。用放射免疫分析法研究了新建立的秋水仙属植物悬浮培养物中秋水仙碱的含量以及各种培养基对该细胞培养物中秋水仙碱产量的影响。酶免疫分析法非常适合定量测定嘉兰和秋水仙属植物种子提取物的高效液相色谱馏分中的秋水仙碱，能够快速、灵敏且精确地测定所研究的物质。初步实验表明，两种秋水仙碱免疫分析法对于组织和细胞培养研究中的秋水仙碱分析、植物提取物分析以及生物合成研究都可能是有用的工具。
