Determination of Azole Pesticides in Human Serum by Coupled Column Reversed-Phase Liquid Chromatography Using Ultraviolet Absorbance and Mass Spectrometric Detection

Abstract

Analysis of some azole pesticides (triadimenol, triadimefon, and tetraconazole) in serum samples was carried out using a coupled column reversed-phase liquid chromatography with ultraviolet detection and liquid chromatography with electrospray mass spectrometry detection. A 5 mm GFF-II, internal surface reversed phase (ISRP, Pinkerton) column coupled to a 3 mm reversed phase, Microsphere C 18 column were used for LC-LC-UV analysis. Linear dynamic ranges were established over more than two orders of magnitude. The applicability of the method to the determination of triadimenol, triadimefon, and tetraconazole in spiked serum samples was evaluated. Recoveries of spiked compounds range between 70.1% and 82.1% and 83.9% and 100.7% at fortification levels of 0.2 and 0.05 mg=mL of each compound, respectively. The relative standard deviation (RSD) is lower than 7.0% in all cases, and the limits of quantitation (LOQ) calculated range from 5 and 10 mg=L.For LC-ES-MS analysis the lineal range, detection limits, precision, and recovery of the analytical method after the clean up step with solid phase extraction using a C 18 cartridge, were determined. The results obtained with both detectors were compared. The methods have been fully validated and utilized for analysis of triadimenol, triamidefon, and tetraconazole in serum samples of five agricultural workers from Almería (Spain).