Multifamily Determination of Phytohormones and Acidic Herbicides in Fruits and Vegetables by Liquid Chromatography–Tandem Mass Spectrometry under Accredited Conditions

Abstract

<jats:p>A 7-min multifamily residue method for the simultaneous quantification and confirmation of 8 phytohormones and 27 acidic herbicides in fruit and vegetables using ultra high-performance liquid chromatography (UHPLC) coupled to tandem mass spectrometry (MS/MS) was developed, validated according to SANTE 12682/2019, and accredited according to UNE-EN-ISO/IEC 17025:2017. Due to the special characteristics of these kinds of compounds, a previous step of alkaline hydrolysis was carried out for breaking conjugates that were potentially formed due to the interactions of the analytes with other components present in the matrix. Sample treatment was based on QuEChERS extraction and optimum detection conditions were individually optimized for each analyte. Cucumber (for high water content commodities) and orange (for high acid and high water content samples) were selected as representative matrices. Matrix-matched calibration was used, and all the validation criteria established in the SANTE guidelines were satisfied. Uncertainty estimation for each target compound was included in the validation process. The proposed method was applied to the analysis of more than 450 samples of cucumber, orange, tomato, watermelon, and zucchini during one year. Several compounds, such as 2,4-dichlorophenoxyacetic acid (2,4-D), 4-(3-indolyl)butyric acid (IBA), dichlorprop (2,4-DP), 2-methyl-4-chlorophenoxy acetic acid (MCPA), and triclopyr were found, but always at concentrations lower than the maximum residue level (MRL) regulated by the EU.</jats:p>

Current Strategies for Studying the Natural and Synthetic Bioactive Compounds in Food by Chromatographic Separation Techniques

“…As it can be observed, liquid chromatography is particularly appropriate for the analysis of polar, non-volatile, and/or thermally labile pesticides. Because of its high selectivity and sensitivity, HPLC and UHPLC in combination with MS/MS have mostly been used in this field especially to determine the insecticides and herbicides belonging to organophosphorus compounds, imidazolinone and pyridine carboxylic acid derivatives, and in study of samples containing multiclass pesticides [16][17][18][19][20][21][22][23][24]27,[30][31][32][33][34]. However, in a few cases, i.e., triazine and phenylurea herbicides, the HPLC coupled with spectrophotometric detection HPLC-UV or DAD has also been applied [25,26,28].…”

“…However, in a few cases, i.e., triazine and phenylurea herbicides, the HPLC coupled with spectrophotometric detection HPLC-UV or DAD has also been applied [25,26,28]. Various kinds of stationary phases (columns) have been used in the HPLC determination of pesticides, mainly C18 [16][17][18]20,[25][26][27][28][29][30][31], and also chiral [19,33], BEH HILIC [21], Hypercarb [22], Obelisc N HILIC [24,34], Acquity UPLC HSS T3 [23,32]. In general, water or water with formic acid or acetic acid or ammonium formate, acetonitrile, and methanol have been applied as mobile phases with gradient or isocratic elution, respectively.…”

“…However, owing to the new SPE (solid phase extraction) systems consisting novel polymers as adsorbents e.g., porous organic polymer Car-DMB, Py-DMB HCP (heterocyclic hypercrosslinked polymer), HPLC analysis further allows the quantification of some pesticides in food samples at concentrations of ng/g [25,28]. As is shown in Table 1, many HPLC-MS/MS techniques with triple quadrupole (TQ), electrospray ionization (ESI) in multiple reaction monitoring (MRM) mode or selected reaction monitoring (SRM) mode have been mainly used for the determination of different kind of pesticides [16,[18][19][20][21]27,31,33]. In addition, the HPLC-MS/MS methods with electrospray ionization (ESI) and quadrupole trap (QTRAP) in multiple reaction monitoring (MRM) mode have been also employed in the analysis of various pesticides [23,24,30,34].…”

“…Studies [19,33] indicate that chiral LC-TQ-ESI-MS/MS and UPLC-TQ-ESI-MS/MS in MRM mode have been successfully applied to the simultaneous enantioselective determination of chiral pesticides in different vegetables and fruits. Martínez et al [27] determined 27 acidic herbicides and 8 phytohormones in fruits and vegetables using UHPLC-TQ-ESI-MS/MS technique in the MRM mode.…”

Detection of Residual Pesticides in Foods

“…In addition to the number of compounds, sample throughput is another key point to be considered when analytical methods are being developed. Grande-Martinez et al [ 2 ] developed a 7 min multifamily residue method for the simultaneous quantification and confirmation of 8 phytohormones and 27 acidic herbicides in fruit and vegetables using ultra high-performance liquid chromatography (UHPLC) coupled to MS/MS. The method, which was validated according to SANTE 12682/2019, was also accredited (UNE-EN-ISO/IEC 17025:2017).…”