Chromatographic simplified multiresidue determination of pesticides in Stevia Rebaudiana (Bertoni) leaves and products. Repetti, M.R. ; Nocetti, M.F.

; Garca S.R.; Vierling J.; De Jess, J.J; Beldomenico H.R. Research and Analysis of Chemical Residues and Contaminants Program (PRINARC FIQ-UNL) Central Laboratory Chemical Engineering Faculty National University of Litoral (FIQUNL) Santiago del Estero 2654 Piso 6, (3000) Santa Fe, Argentina. e-mail: mrepetti@fiq.unl.edu.ar The culture Stevia Rebaudiana (Bertoni) is of increasing relevance because it recognized pharmacological and nutritional properties, especially (specially es ms informal) as a source of non-caloric natural sweeteners, like the derived high purity Rebaudioside A (rebiana) and other steviol glycosides. An increasing amount of countries have introduced very recently regulations allowing the use of Stevia and derivates as food ingredients, after a long time of restrictions owing to safety and toxicological controversies about several components [1]. It represents a culture of great potential, innovative and profitable, specially for in Latin-American countries (the plant is of South American origin Paraguay, 1887-), because the growing demand of their products for many uses (cosmetic, agro-technical, pharmaceutical, food industry, etc.), especially as a natural alternative to the controversial synthetic sweeteners, in food, drink and beverages formulations. Argentinean production, located in north-east provinces (Misiones, Corrientes, Formosa, and incipiently in other provinces) enhances the interest in promoting products both for domestic and export destination. There is few literature on pesticide residues in Stevia and derivates, no LMRs or specific regulations on residues were found in our search. Despite this culture do not demand a great load of agrochemicals, it is important to consider the influence of the agro-technical practice applied to the rest of crops in the same region of production. This scenario attracted our attention, and led to work on up-to-date multiresidue methods to deal with a complex vegetal matrix like leaves of Stevia (Stevia rebaudiana Bertoni, native variety), that naturally contents many nutrients, flavonoids, esterols, terpenes, essential oils and chlorophyl, besides the steviol gycosides (7-13 % of total weight of leaves)[2]. In this case experimentation of several types and procedure conditions of clean-up on dry leaves, led to the adoption of a QuEChERS based strategy, with acetonitrile and non-dispersive florisil/PSA treatment, suitable for determination of endosulfan (, , sulfate) and bifenthrin by GC ECD. Similar cleanup was also studied and selected for the determination of 40 representative pesticides (considering variety of chemical class and use relevance) with final determination by UHPLC MSMS. Principal figures of merit including linearity, matrix effects and recovery studies were satisfactory and are presented here. The methodology was applied to dry leaves commercial products from the three mentioned Argentinean provinces. Samples of extracted powder products (rebaudioside and stevioside based sweeteners), available in domestic market, were submitted to the study resulting a very less problematic sample. No positive results were found in samples studied and satisfactory applications of developed methodology were achieved, opening to further studies on this culture, with high potential for future production and international trading.

[1] Carakostas M.C., Curry L.L., Boileau A.C., Brusick D.J. (2008). Overview: The history, technical function and safety of rebaudioside A, naturally occurring steviol glycoside, for use in food and beverages. Food and Chemical Toxicology 46, S1 S10. [2] Nocetti, M.F. (2011) Application of a multiresidue method for the determination of pesticides in Stevia rebaudiana (Bertoni) leaves using chromatographic techniques. Thesis Graduate Chemistry Degree of Chemical Engineering Faculty, National University of Litoral (FIQ-UNL), Santa Fe, Argentina.