Evolution of a specific fluorogenic derivatization of ivermectin for bioanalytical applications. A review
The evolution of the fluorogenic derivatization of ivermectin is traced through a series of continual modifications that have resulted in improvements in speed and sensitivity. Since the original development of this selective analytical technique, the reaction time has been shortened from 24 h at 100 °C to <30 s at room temperature and, through modifications of the derivatization reagent and catalyst, the sensitivity has also been increased 50-fold to 20 pg of analyte with no significant decrease in precision. A procedure is reported, based on the use of fluorescence derivatization, which eliminates the use of solid-phase columns for sample preparation and fluorophore isolation, and is faster and less cumbersome than previous methods. The method was evaluated with cattle and canine plasma samples over the concentration range 1.0–40 ng ml–1 of ivermectin. It has an accuracy of 1.9%(mean relative error) over this concentration range and a precision of 5.6%(RSD) at the 1 ng ml–1 ivermectin concentration level in a 1 ml plasma sample.