Pesticide Residues In Olive Oil Research Articles

Determination of 222 pesticide residues in olive oil by fully automatic QuEChERS pre-treatment instrument coupled with gas chromatography-quadrupole-time-of-flight mass spectrometry

Pesticide residues may be present in olive oil because pesticides are applied to olive trees during their cultivation and growth for pest prevention and some of these pesticides are not easily degraded. Studies on pesticide residues in olive oil have mainly focused on the detection of single types of pesticide residues, and reports on the simultaneous detection of multiple pesticide residues are limited. At present, hundreds of pesticides with different polarities and chemical properties are used in practice. In this study, an analytical method based on fully automatic QuEChERS pretreatment instrument coupled with gas chromatography-quadrupole time-of-flight mass spectrometry (GC-QTOF-MS) was established for the rapid determination of 222 pesticide residues in olive oil. The effects of acetonitrile acidification concentration, n-hexane volume, oscillation time, centrifugation temperature, and purification agent on the determination of the 222 pesticide residues were investigated. First, ions with good responses and no obvious interference were selected for quantification and characterization. The purification process was then developed by setting the parameters of the fully automatic QuEChERS pretreatment instrument to optimal values. The sample was extracted with acetonitrile containing 2% formic acid, and the supernatant was purified by centrifugation in a centrifuge tube containing 400 mg N-propylethylenediamine (PSA), 400 mg octadecylsilane-bonded silica gel (C18), and 1200 mg anhydrous magnesium sulfate. The purified solution was blown dry with nitrogen and then fixed with ethyl acetate for instrumental analysis. Finally, a matrix standard solution was used for quantification. The method was validated in terms of matrix effects, linear ranges, limits of detection (LODs) and quantification (LOQs), accuracies, and precisions. The results showed that 86.04% of the 222 pesticides had linear ranges of 0.02-2.00 μg/mL, 10.81% had linear ranges of 0.10-2.00 μg/mL, and 3.15% had linear ranges of 0.20-2.00 μg/mL. The pesticide residues showed good relationships within their respective linear ranges, and the correlation coefficients (R2) were greater than 0.99. The LODs of all tested pesticides ranged from 0.002 to 0.050 mg/kg, and their LOQs ranged from 0.007 to 0.167 mg/kg. Among the 222 pesticides determined, 170 pesticides had LOQs of 0.007 mg/kg while 21 pesticides had LOQs of 0.017 mg/kg. At the three spiked levels of 0.2, 0.5, and 0.8 mg/kg, 79.58% of all tested pesticides had average recoveries of 70%-120% while 65.92% had average recoveries of 80%-110%. In addition, 93.54% of all tested pesticides had relative standard deviations (RSDs, n=6)<10% while 98.35% had RSDs (n=6)<20%. The method was applied to 14 commercially available olive oil samples, and seven pesticides were detected in the range of 0.0044-0.0490 mg/kg. The residues of fenbuconazole, chlorpyrifos, and methoprene did not exceed the maximum limits stated in GB 2763-2021. The maximum residual limits of molinate, monolinuron, benalaxyl, and thiobencarb have not been established. The method utilizes the high mass resolution capability of TOF-MS, which can improve the detection throughput while ensuring good sensitivity. In addition, high-resolution and accurate mass measurements render the screening results more reliable, which is necessary for the high-throughput detection of pesticide residues. The use of a fully automatic QuEChERS instrument in the pretreatment step reduces personnel errors and labor costs, especially when a large number of samples must be processed, thereby offering significant advantages over other approaches. Moreover, the method is simple, rapid, sensitive, highly automatable, accurate, and precise. Thus, it meets requirements for the high-throughput detection of pesticide residues in olive oil and provides a reference for the development of detection methods for pesticide residues in other types of oils as well as the automatic pretreatment of complex matrices.

Determination of Pesticide Residues in Olive Oil Using QuEChERS Extraction and Liquid Chromatography–Orbitrap High-Resolution Mass Spectrometry: Comparison of Different Clean-Up Sorbents and Validation Study

The aim of this study was the optimization of the clean–up step in the widely applied QuEChERS method for the determination of 39 representative multiclass pesticides in olive oil with Ultra-High-Performance Chromatography–Orbitrap Mass Spectrometry (UHPLC-Orbitrap-MS). The analytical methodology combines the original version of QuEChERS extraction with two different clean-up-step approaches, using firstly a combination of Z-Sep+, PSA and MgSO4 and secondly EMR-lipid. The methods were compared for their efficiency in the removal of fats and co-extractives and their effect on the analytical performance characteristics. Both methods were evaluated in terms of linearity, matrix effects (ME), recovery, precision, limits of detection (LOD) and quantification (LOQ) and expanded uncertainty in three spiking levels of 30, 100 and 300 μg/kg. The recoveries ranged between 70–113% for 95% of analytes (RSDr &lt; 14%) when EMR-lipid was used as a sorbent, while in the case of Z-Sep+/PSA/MgSO4 recoveries ranged between 72–107% for 92% of analytes (RSDr &lt; 18%). ME showed low signal suppression for 77% of analytes in the case of Z-Sep+/PSA/MgSO4 and for 85% of analytes in the case of EMR-lipid. According to the results, both methodologies provided good analytical performances fulfilling validation criteria; however, the EMR-lipid sorbent showed better clean-up capacity (i.e., less matrix effects and lower variability in extraction recoveries) and validation parameter values for more analytes. The validated method was successfully applied to 30 olive oil samples from different regions of Greece. No residues have been identified in the analyzed samples.

A Quick and Inexpensive Method to Determine Pesticide Residues in Olive Oil by UHPLC-MS/MS and GC-MS/MS

Olive oil samples were analyzed for twenty-nine pesticides by means of gas chromatography and Ultra HPLC coupled to mass spectrometry. The list of the investigated pesticide residues includes the twenty-seven pesticide residues of the target protocol of COIPT proficiency tests, yearly organized in our laboratory.

Results of the 16th proficiency test on the determination of pesticide residues in olive oil

The Italian National Reference Laboratory (NRL) for pesticide residues yearly organizes proficiency tests (PTs) on olive oil in cooperation with the International Olive Council, addressed to European laboratories (NRLs, official control laboratories, and private laboratories), involved in the National and European monitoring programs. The main aim of these PTs is to compare the performances of the laboratories in Mediterranean and European countries and to promote mutual acceptance of pesticide residue data regarding the analytical controls of olive oil. In this paper, a description of the last PT named COIPT-16 is presented. The test material consisted of a commercial olive oil spiked with six different pesticides. The assigned value of each analyte was determined as a consensus value based on the results of the participants calculated using Algorithm A of the ISO 13528:2015. As relative target standard deviation in the z score formula, the 25 % of the robust mean value was considered. The global performance of each participating laboratory was assessed by calculating the average of the squared z scores only for laboratories that have detected at least 80 % of the analytes present in sample achieving the sufficient scope as indicated in the initial protocol. Two compounds, alpha-endosulfan and beta-endosulfan have been analyzed again in this PTas in the previous PT to compare the performances of the laboratories whose supplied results both PTs. The quick, easy, cheap, effective, rugged, and safe methodology was used by the majority of laboratories corresponding to 64 % of participants.

Dual-layer solid-phase extraction based on molecular imprinting technology: Seeking a route to enhance selectivity for trace analysis of pesticide residues in olive oil.

Aiming to introduce a multiresidue analysis for the trace detection of pesticide residues belonging to organophosphorus and triazine classes from olive oil samples, a new sample preparation methodology comprising the use of a dual layer of "tailor-made" molecularly imprinted polymers (MIPs) SPE for the simultaneous extraction of both pesticides in a single procedure has been attempted. This work has focused on the implementation of a dual MIP-layer SPE procedure (DL-MISPE) encompassing the use of two MIP layers as specific sorbents. In order to achieve higher recovery rates, the amount of MIP layers has been optimized as well as the influence of MIP packaging order. The optimized DL-MISPE approach has been used in the preconcentration of spiked organic olive oil samples with concentrations of dimethoate and terbuthylazine similar to the maximum residue limits and further quantification by HPLC. High recovery rates for dimethoate (95%) and terbuthylazine (94%) have been achieved with good accuracy and precision. Overall, this work constitutes the first attempt on the development of a dual pesticide residue methodology for the trace analysis of pesticide residues based on molecular imprinting technology. Thus, DL-MISPE constitutes a reliable, robust, and sensitive sample preparation methodology that enables preconcentration of the target pesticides in complex olive oil samples, even at levels similar to the maximum residue limits enforced by the legislation.

Uncertainty estimation in the analysis of pesticide residues in olive oil using data from proficiency tests

In this work we report the results for estimating the measurement uncertainty (MU) following up the application of two different approaches, relatively the top-down procedure, by using proficiency test data. We have focused the estimation on the olive oil matrix. We used the analytical data obtained from five selected editions of the Proficiency Tests (PTs, from 2007 to 2011) on pesticide residues in olive oil to estimate the MU. These PTs have been organized by Istituto Superiore di Sanità annually in cooperation with International Olive Council (IOC) since 1997. The number of participants in each trial ranged from 10 to 43. We used a total of 34 pesticide results. The expanded uncertainty U (c) was calculated using a covering factor k = 2 for a confidence interval of 95%. In the approach 1, the within–laboratory reproducibility standard deviation is combined with estimates of the method and laboratory bias using PTs data. In the approach 2, the way of estimating the MU is based only on the bias that the laboratory has obtained participating in a sufficient number of the IOC proficiency tests. Comparing the relative expanded uncertainty based on these different approaches we notice values quite constant and close, from 42% to 48%. Moreover, these calculated expanded uncertainties are less than the default value of 50% (corresponding to a 95% confidence level), adopted from European guidance document SANCO based on the fit-for-purpose relative standard deviation (FFP-RSD).

DETERMINATION OF ORGANOPHOSPHORUS PESTICIDE RESIDUES IN OLIVES GROWN IN BURSA, TURKEY

The use of pesticides in agriculture has been continuously increasing in recent years and has led to an increase in world food production. Organophosphorus compounds are among the most widely used pesticides in the world. Because of their long half-life time, they contaminate soils as well as surface and ground water. In addition, these residues in food constitute a significant health risk. Olive and its products are extensively consumed foods in the Mediterranean countries. However, olive trees are attacked by several pests and diseases, making it necessary to apply pesticides to ensure crop protection, which can leave residues on the fruit. Because pesticide residues in food constitute a significant health risk and olive oil has a high consumption rate among people of the producing countries, the continuous control of pesticide residues in olive oil is of great importance. In this study, the occurrence of nine organophosphorus pesticide residues in olives (Olea europaea L. ‘Gemlik’) in Mudanya and Gemlik towns (Bursa, Turkey; latitude: 29°04’E, longitude: 40°39’N) was investigated. Olive samples in the black maturation stage from twenty orchards were analyzed for pesticide residues using different extraction methods by capillary gas chromatography (GC) using NPD with a capillary column. Of the eight organophosphorus pesticides investigated, four were detected in olives namely azinphos-ethyl, chlorpyrifos, chlorpyrifos-methyl, and carbophenothion. The other four, diazinon, methidathion, malathion, and pirimiphos-methyl, were below detection limits. All samples contained more than one pesticide and 17 were found to contain chlorpyrifos at concentrations slightly above the maximum pesticide residue limit (100 µg kg-1, MRL).

Use of absorbent materials in on-line coupled reversed-phase liquid chromatography–gas chromatography via the through oven transfer adsorption desorption interface

The use of absorbents as retaining materials in the through oven transfer adsorption desorption interface (TOTAD) of an on-line coupled reversed-phase liquid chromatography–gas chromatography system (RPLC–GC) is proposed for the first time. A comparative study of an adsorbent (Tenax TA) and two absorbents, namely polydimethylsiloxane and poly(50% phenyl/50% methylsiloxane) is performed to establish the best experimental conditions for the automated and simultaneous determination of 15 organophosphorus and organochlorine pesticide residues in olive oil. The proposed method provides satisfactory repeatability (RSDs lower, in general, than 8.5%) and sensitivity (limits of detection ranging from 0.6 to 81.9 μg/L) for the investigated compounds.

Development of an immunosensor based on the measurement of fluorescence

Consumers concern about the presence of pesticides in food is increasing. However current techniques for detecting pesticide residues in olive oil are very expensive and slow, which make them difficult to apply in commercial conditions. For this reason, the Instituto Valenciano de Investigaciones Agrarias (IVIA) and Universidad Politecnica de Valencia (UPV) have decided to develop an immunosensor as an alternative, fast and cheap method to measure the concentration of some pesticides in olive oil. This work shows the developed transducer by IVIA based on a photovoltaic sensor that transforms the fluorescent radiation to an electrical signal. The immunosensor is based on an immunologic reaction which causes fluorescence on the sample. The intensity of the fluorescence is related with the concentration of marked antibodies. The performed tests with two types of immunoassays and fluorescent markers (FITC and ELF97) have demonstrated the robustness and reliability of the developed transducer. The immunosensor is able to discriminate different concentrations of marked antibodies. This study has demonstrated the possibility of applying this immunosensor to detect chemical residues from pesticide treatments in olive oil samples. These results validate the sensor for future commercial applications.

Application of Gas Chromatography Coupled to Triple Quadrupole Mass Spectrometry for the Multiresidue Analysis of Pesticides in Olive Oil

A new multiresidue method has been developed and validated for the simultaneous determination of 100 pesticide residues in olive oil. The determination of pesticide residues was carried out in only 19 min by gas chromatography coupled to tandem mass spectrometry using a triple quadrupole mass analyzer. The mass spectrometer was operated in electron ionization and the selection reaction monitoring mode was used, acquiring two or three fragmentation reactions per compound. Two extraction processes were studied, and an evaluation of the stability and sensitivity of the chromatographic system has been performed for the tested extraction procedures. The final proposed methodology was based on a liquid-liquid partition with an n-hexane/acetonitrile mixture followed by a gel permeation chromatography cleanup step. An adequate lineal relation was obtained in the studied concentration range (10-200 microg kg (-1)); the recovery values were in the range 70-110% for the two levels of concentration studied: 12 and 50 microg kg (-1). Precision values, expressed as relative standard deviation, were lower than 18% at the aforementioned spiking levels; detection limits, confirmation limits, and quantitation limits were below or equal to 1.9, 2.6, and 3.6 microg kg (-1), respectively. The developed methodology was applied to the analysis of pesticide residues in real samples of olive oil from the south of Spain.

Application of high-performance liquid chromatography–tandem mass spectrometry with a quadrupole/linear ion trap instrument for the analysis of pesticide residues in olive oil

This article describes the development of an enhanced liquid chromatography-mass spectrometry (LC-MS) method for the analysis of pesticides in olive oil. One hundred pesticides belonging to different classes and that are currently used in agriculture have been included in this method. The LC-MS method was developed using a hybrid quadrupole/linear ion trap (QqQ(LIT)) analyzer. Key features of this technique are the rapid scan acquisition times, high specificity and high sensitivity it enables when the multiple reaction monitoring (MRM) mode or the linear ion-trap operational mode is employed. The application of 5 ms dwell times using a linearly accelerating (LINAC) high-pressure collision cell enabled the analysis of a high number of pesticides, with enough data points acquired for optimal peak definition in MRM operation mode and for satisfactory quantitative determinations to be made. The method quantifies over a linear dynamic range of LOQs (0.03-10 microg kg(-1)) up to 500 microg kg(-1). Matrix effects were evaluated by comparing the slopes of matrix-matched and solvent-based calibration curves. Weak suppression or enhancement of signals was observed (<15% for most-80-of the pesticides). A study to assess the identification criteria based on the MRM ratio was carried out by comparing the variations observed in standard vs matrix (in terms of coefficient of variation, CV%) and within the linear range of concentrations studied. The CV was lower than 15% when the response observed in solvent was compared to that in olive oil. The limit of detection was < or =10 microg kg(-1) for five of the selected pesticides, < or =5 microg kg(-1) for 14, and < or =1 microg kg(-1) for 81 pesticides. For pesticides where additional structural information was necessary for confirmatory purposes-in particular at low concentrations, since the second transition could not be detected-survey scans for enhanced product ion (EPI) and MS3 were developed.

Automated determination of pesticide residues in olive oil by on-line reversed-phase liquid chromatography–gas chromatography using the through oven transfer adsorption desorption interface with electron-capture and nitrogen–phosphorus detectors operating simultaneously

A rapid method for the multiresidue analysis of pesticides in olive oil is presented. Pesticides are analyzed by on-line coupling reversed-phase liquid chromatography–gas chromatography using the through oven transfer adsorption desorption (TOTAD) interface with subsequent simultaneous electron-capture and nitrogen–phosphorus detection by post-column splitter. An autosampler is employed and the olive oil is simply filtered before the chromatographic analysis. Organophosphorus, organochlorine and triazine pesticides are determined in one run. The limits of detection are below the required maximum residue levels and calibration curves are linear in the range tested. Repeatabilities (intra-day and inter-days) are good. The method was satisfactory applied to the routine analysis of numerous olive oil samples.

Determination of pesticide residues in olive oil and olives

Pesticide-residue determination in olive oil is becoming a very important and challenging issue. The present article provides a review of the most significant methods for determining pesticides in olive oil. The discussion of sample-treatment procedures is focused on not only the final product (olive oil), but also the direct analysis of the raw material (olive fruits). We outline extraction and clean-up procedures (e.g., liquid-liquid partition, gel-permeation chromatography (GPC), solid-phase extraction (SPE), matrix solid-phase dispersion (MSPD) and Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS). However, we pay specially attention to the clean-up stage, which plays an important role in the whole process. In addition, identification and quantitation of pesticides have been carried out by gas chromatography (GC) coupled to different detectors (flame-ionization (FID), nitrogen-phosphorus (NPD) and electron-capture (ECD), although either GC-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode or GC-tandem MS (GC-MS 2) are becoming the combination of choice, because of higher selectivity and confirmation capabilities. In recent years, the appearance of new, more polar pesticides has prompted the use of liquid chromatography-MS (LC-MS) instead of GC-MS, and we outline its advantageous features for the analyses of polar pesticides in olive oil.

Automated multiresidue analysis of pesticides in olive oil by on-line reversed-phase liquid chromatography–gas chromatography using the through oven transfer adsorption–desorption interface

A multiresidue, automated and rapid method for the determination of pesticide residues in olive oil is presented. The method employs the through oven transfer adsorption–desorption interface for the on-line coupling of reversed-phase liquid chromatography and gas chromatography. In this fully automated system, olive oil is directly injected with no sample pre-treatment step other than filtration. Methanol–water is used as eluent in the liquid chromatography pre-separation step. The selected liquid chromatography fraction containing the pesticides is automatically transferred to the gas chromatography. The liquid chromatography column flow during elution is different from the flow during the transfer. Using a flame ionisation detector, pesticide detection limits varied from 0.1 to 0.3 mg/l.

Direct analysis of pesticide residues in olive oil by on-line reversed phase liquid chromatography-gas chromatography using an automated through oven transfer adsorption desorption (TOTAD) interface.

A fully automated on-line reversed phase liquid chromatography-gas chromatography system is described. The system uses a prototype of the automated through oven transfer adsorption desorption interface. The system is demonstrated by presenting a new rapid method for the determination of pesticide residue in olive oil, which is injected directly with no sample pretreatment step other than filtration. Methanol:water is used as the eluent in the LC preseparation step, while the LC fraction containing the pesticide is automatically transferred to the gas chromatograph. Detection limits of pesticides varied from 0.18 to 0.44 mg/L when a flame ionization detector was used. As an example, relative standard deviation and linear calibration are presented for terbutryne.

Determination of residual pesticides in olive oil by GC-MS and HPLC-MS after extraction by size-exclusion chromatography.

This work describes the development of a method for analyzing pesticide residues in olive oil by GC-MS and HPLC-MS. Pesticides were separated from the oily matrix by size-exclusion chromatography. After extraction, 20 pesticides were separated and analyzed by GC-MS and 11 others HPLC-MS in electrospray mode. The development of this method enabled us to identify and quantify the pesticides of interest.

Pesticide residues in olive oil.

The attacks of pests and diseases and the presence of weeds make it necessary to apply pesticides to olive trees to ensure crop protection. Residues of these compounds may remain and contaminate the oil produced. For the analysis of pesticide residues in olive oil, the most common methods are multiresidue methods for fatty substrates, based on partitioning between hexane or light petroleum and acetonitrile. Recently, other methods have been applied, such as ready-to-use, disposable minicolumns or direct injection of oil into a capillary gas chromatograph equipped with a precolumn with an oil recovery tank. Although several pesticides are registered in oil-producing countries for use on olive trees, available literature on the level and fate of residues is very limited. However, it is clear that fat-soluble pesticides tend to concentrate in the oil, both after full coverage and bait spraying, and their use close to harvest should therefore be avoided. Because it is sometimes necessary to use such pesticides late in autumn because of their effectiveness in cases of severe attack, residue trials should be carried out to determine the residue concentration in oil and to set a reasonable preharvest safety interval. Data produced by such trials would permit the establishment of MRLs (tolerances) in olive oil to cover cases where the residues, although relatively high, are not of toxicological significance for consumers (risk assessment). Such is the case with corn oil and the fat-soluble insecticide methyl pirimiphos, registered in the U.S. for use on corn. The U.S. EPA tolerance for methyl pirimiphos in corn is 8 mg/kg, whereas it is 11 times higher (88 mg/kg) for corn oil because it is known to concentrate in the oil. Similar provisions for olive oil, based on data from residue trials according to Good Agricultural Practice, the long-term toxicity of each pesticide as expressed by its ADI for man, and olive oil consumption patterns, would facilitate international trade of this commodity. On the other hand, because of the high dietary and health value of olive oil, it is desirable that toxic pesticide residues be kept as low as possible. Therefore, it would be preferable not to rely only on chemical pest control treatments but to develop and apply alternative plant protection techniques such as Integrated Pest Management (IPM).(ABSTRACT TRUNCATED AT 400 WORDS)