Colchicine Determination in Gloriosa spp. by HPLC

Abstract

Colchicine is a phytochemical alkaloid with important pharmaceutical, biomedical, and agricultural properties. An HPLC method for colchicine analysis in Gloriosa spp. extract was developed in this study. The ideal compound separation and detection conditions were determined to be as follows: C18 column, with 50 mM KH₂PO₄ water: acetonitrile ratio of 40:60 in a mobile phase at a flow rate of 1 mL.min^-1. Colchicine was detected at 254 nm, with the linearity of the standard curve in the range of 20-100 μg.mL^-1. The developed method was precise with a value of 0.16 %RSD intra-day and 0.93 %RSD inter-day which was lower than 1%RSD. The limit of detection (LOD) and limit of quantification (LOQ) were 4.98 and 62.23 ng.mL^-1, respectively. Accuracy was evaluated using recovery (%) value, which yielded an average of 99.41 % recovery. The specificity analysis showed that interference was not observed at the retention time of colchicine (4.4 min). The method was stable without effects from mobile phase ratio, flow rate, and column temperature. This HPLC method was used to quantify colchicine from various Gloriosa spp. samples, the content was in the range of 117.97 - 380.66 μg.gDW^-1. The conditions and protocols used in this study have been rigorously modified to produce favorable outcomes for colchicine. Thus, this suggests that this approach can be used as a standardized procedure for further colchicine analysis, especially in Gloriosa spp. which will benefit studies in pharmaceutical science and lead to further developments.