Determinação de fungicidas em sucos de laranja empregando método de extração mini-luke modificado e UPLC-MS/MS

Abstract

Orange juice is a relevant contribution to the Brazilian economy, as long as five glasses of orange juice consumed in the world, three are produced in Brazil. For the purpose of increase the production of fruits and, consequently, juices, there is a growing concern about the presence of pesticide residues in food, so monitoring by analytical methods is required in order to ensure consumer s health. This study shows the validation of an analytical method for the determination of nine fungicides in natural and processed orange juices by UPLC-MS/MS. The extraction procedure used was the modified mini-Luke method. Developed in 1975, it was modified in later years and has been used to monitor the presence of pesticides in non-fatty matrices such as fruits and vegetables. The method consists of adding 30 mL of acetone followed by 60 mL of 1:1 (v/v) mixture of petroleum ether and dichloromethane to 15 g of sample. After homogenization and centrifugation, an aliquot of 1.2 mL was transferred into tubes and evaporation led to a water bath to near dryness. The residue was reconstituted in 1 mL of methanol containing 0.1% acetic acid and analyzed by UPLC-MS/MS. The parameters evaluated for the validation were: calibration curve and linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy (as recovery%), precision (as RSD %) and selectivity (as matrix effects%). The results were satisfactory for all parameters, thus showing that the modified mini-Luke extraction method was effective to extract the studied fungicides in natural and processed orange juices. In the range of concentration studied, the method was linear. The method LOQ were 10, 20 and 50μg kg-1 for six, two and one fungicides studied, respectively. It is noted that these values are the same or below the values of the maximum residue limits (MRL) established by Brazilian, American and European laws, indicating that the method can be used worldwide to determine the nine fungicides studied in orange juice.It can be inferred that the method is selective, as long as this parameter was assessed from the matrix effects. It was negative for most of the fungicides studied and all values remained in the range -20 to +20%, not influencing the results. The analyzed fungicides, show average recoveries between 70 and 120%, only excepting thiophanate - methyl and carbendazin. The thiophanate-methyl presented values below 70% when spiked at concentrations of 10 and 20 μg kg 1 and carbendazim presented values above 120% when spiked at concentrations of 10 μg kg 1. The extraction method, combined with modern chromatographic technique for the determination of analytes, were effective for the analysis of pesticide residues in orange juice.