HIGH PERFORMANCE LIQUID CHROMATOGRAPHY AND ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) FOR THE DETERMINATION OF COLCHICINE AND RELATED COMPOUNDS

Document Type : Original Article

Authors

1 Department of Pharmacognosy, Faculty of Pharmacy, University of Zagazig, Egypt.

2 Institute of Pharmaceutical Biology, Munich University, Munich, Germany

3 Dept. of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig , Egypt

4 Department of Pharmacognosy, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt

Abstract

An HPLC method was applied for the determination of colchicine and colchicoside in the methanolic extract of the seeds of Colchicum autumnale L. during different stages of seed maturity. The method is carried out isocratically on a reversed-phase column using H2O-CH3CN (95:5) +100 µl H3PO4. The detection limits were 0.10 and 0.05 µg for colchicine and colchicoside, respectively and the recoveries (99.2 ± 1.1%) were high and quantitative. Also, a highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) was developed for detection and quantitative determination of colchicine, its structurally related compounds (colchicinoids) in seeds of C. autumnale and suspension culture of Colchicum variegatum L. A combined experiment (HPLC- ELISA) was performed for detection of colchicine and some other colchicinoids in C. autumnale seeds extract.

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