Application of matrix solid-phase dispersion and liquid chromatography–mass spectrometry to fungicide residue analysis in fruits and vegetables

A method based on matrix solid-phase dispersion (MSPD) and liquid chromatography-electrospay ionization-mass spectrometry used to analyze fifteen fungicide residues in fruits and vegetables is described. The method required only 0.5 g of sample, C(18)-bonded silica was used as dispersant sorbent, and ethyl acetate was used as eluting solvent. Fortified recoveries in apple, orange, banana, lettuce, grape and tomato samples ranged from 71% to 102% and relative standard deviations were less than 13% with fortified levels of 0.03-1.5 mg kg(-1). Detection and quantification limits were 1 approximately 30 microg kg(-1) and 4 approximately 100 microg kg(-1), respectively, with linear calibration curves extending up to 15 mg kg(-1). The analytical characteristics of MSPD compared very favorably with those found for a classical multiresidue method: the quick, easy, cheap, effective, rugged, and safe (QuEChERS) method. The method was applied to determine the fungicides in real samples. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) was used as confirmatory tool for positive samples.     

Simultaneous determination of eight pesticide residues in coconut using MSPD and GC/MS

A simple and effective extraction method based on matrix solid-phase dispersion (MSPD) was developed to determine dimethoate, malathion, lufenuron, carbofuran, 3-hydroxycarbofuran, thiabendazole, difenoconazole and trichlorfon in coconut pulp using gas chromatography-mass spectrometry with selected ion monitoring (GC/MS, SIM). Different parameters of the method were evaluated, such as type of solid-phase (C(18), alumina, silica-gel and Florisil), the amount of solid-phase and eluent (dichloromethane, acetone, ethyl acetate, acetonitrile, n-hexane and n-hexane:ethyl acetate (1:1, v/v)). The best results were obtained using 0.5 g of coconut pulp, 1.0 g of C(18) as dispersant sorbent, 1.0 g of Florisil as clean-up sorbent and acetonitrile saturated with n-hexane as eluting solvent. The method was validated using coconut pulp samples fortified with pesticides at different concentration levels (0.25-1.0 mg kg(-1)). Average recoveries (four replicates) ranged from 70.1% to 98.7%, with relative standard deviations between 2.7% and 14.7%, except for lufenuron and difenoconazole, for which recoveries were 47.2% and 48.2%, respectively. Detection and quantification limits for coconut pulp ranged from 0.02 to 0.17 mg kg(-1) and from 0.15 to 0.25 mg kg(-1), respectively.     

水果蔬菜中多种杀菌剂的基质固相分散液相色谱测定方法研究

采用基质固相分散(MSPD)代替液液分配和固相萃取,从蔬菜水果中提取、净化10种常用杀菌剂农药残留,用C18硅胶交联剂作为固相吸附剂,乙酸乙酯作为洗脱液,用HPLC/PDA和LC-MS进行分析检测。10种杀菌剂在0.5~5 mg/kg含量的添加回收率在65%~110%之间,相对标准偏差小于10%,使用HPLC、PDA和LC-MS的检出限分别在0.02~0.2 mg/kg和0.002~0.01 mg/kg之间。该方法节省溶剂,提取和净化一步完成,适用于新鲜水果和蔬菜中10种杀菌剂的残留分析。     

Application of gas chromatography-triple quadrupole mass spectrometry in the quantification-confirmation of pesticides and polychlorinated biphenyls in eggs at trace levels

A new multiresidue method has been developed and validated for the simultaneous analysis of 57 compounds, including organochlorine and organophosphorus pesticide residues (OCPs and OPPs) and polychlorinated biphenyls (PCBs), in eggs at trace levels by gas chromatography coupled to triple quadrupole mass spectrometry (GC-QqQ-MS/MS). Egg samples were extracted by a simple and fast matrix solid phase dispersion (MSPD) procedure using C18 as sorbent, and ethyl acetate and acetonitrile saturated in n-hexane (85:15, v/v) as elution solvent with a simultaneous clean up with Florisil in-line. The QqQ analyzer acquired data in selected reaction monitoring (SRM) mode, permitting both quantification and confirmation in a single injection with a running time reduced up to 17.70 min. Recovery was in the range of 70-110% and 70-106% at 15 and 50 microg/kg, respectively. Precision values expressed as relative standard deviation (RSD) were lower than 20%. Linearity in the range of 10-150 microg/kg provided determination coefficients (R(2)) higher than 0.98 for all compounds. Limits of detection (LODs) for pesticides were < or =2.25 microg/kg and limits of quantification (LOQs) ranged from 0.02 to 7.78 microg/kg. LODs for PCBs were < or =0.41 microg/kg and LOQ were < or =0.71 microg/kg. The method was applied to real samples. Endosulfan sulphate and p,p'-DDE were found in two samples at concentrations below the first calibration level.     

MSPD procedure for determining buprofezin,tetradifon, vinclozolin,and bifenthrin residues in propolis by gas chromatography–mass spectrometry

A simple and effective extraction method based on matrix solid-phase dispersion (MSPD) was developed to determine bifenthrin, buprofezin, tetradifon, and vinclozolin in propolis using gas chromatography–mass spectrometry in selected ion monitoring mode (GC–MS, SIM). Different method conditions were evaluated, for example type of solid phase (C₁₈, alumina, silica, and Florisil), the amount of solid phase and eluent (n-hexane, dichloromethane, dichloromethane–n-hexane (8:2 and 1:1, v/v) and dichloromethane–ethyl acetate (9:1, 8:2 and 7:3, v/v)). The best results were obtained using 0.5 g propolis, 1.0 g silica as dispersant sorbent, 1.0 g Florisil as clean-up sorbent, and dichloromethane–ethyl acetate (9:1, v/v) as eluting solvent. The method was validated by analysis of propolis samples fortified at different concentration levels (0.25 to 1.0 mg kg⁻¹). Average recoveries (four replicates) ranged from 67% to 175% with relative standard deviation between 5.6% and 12.1%. Detection and quantification limits ranged from 0.05 to 0.10 mg kg⁻¹ and 0.15 to 0.25 mg kg⁻¹ propolis, respectively.     

底物固相分散法测定土壤中甲氰菊酯残留量

以甲氰菊酯为分析对象，将一种新型的样品预处理技术-底物固相分散法(MSPD)应用于测定土壤中农药残留。确定了MSPD法的实验条件：土壤量为4g，水的加入量为1mL，固相吸附剂弗罗里硅土的用量为10．0g，淋洗剂为15mL石油醚-乙酸乙酯(1 9)。土壤样品在此条件下处理后无需进一步净化即可用气相色谱／电子捕获检定器测定。三种土壤的三种加标水平的回收率均在90％以上，相对标准偏差小于5％(n＝5)，最小检出质量比为0．002mg/kg。     

基质固相分散-气相色谱/质谱法测定蔬菜中的邻苯二甲酸酯

利用基质固相分散-气相色谱/质谱法测定了蔬菜中的邻苯二甲酸二甲酯、邻苯二甲酸二乙酯、邻苯二甲酸二丁酯、邻苯二甲酸丁基苄基酯和邻苯二甲酸二异辛酯。蔬菜样品经弗罗里硅土和石墨化炭黑研磨均匀后,用乙酸乙酯淋洗净化,结果表明:上述5种邻苯二甲酸酯在0.05~10.00 mg/L 范围内具有良好的线性,样品的添加回收率为76%~90%,相对标准偏差为2%~7%,5种邻苯二甲酸酯的检出限为0.01~0.024 mg/kg。该方法操作简便、经济,分析速度快,适用于大批量样品的分析。     

Determination of microcystins in biological samples by matrix solid-phase dispersion and liquid chromatography-mass spectrometry

A method for the detection and quantification of the microcystins (MCs)-MC-LR, MC-RR and MC-YR-in biological samples by matrix solid-phase dispersion (MSPD) has been developed. The optimum extraction conditions were 500 mg of liver or kidney, C18 bonded silica as dispersant, and a mixture methanol-water (70:30) as eluent. The MCs were determined by liquid chromatography electrospray mass spectrometry (LC/ES/MS). Recoveries of biological extracts at three different spiked levels (1-10 mg kg(-1)) ranged from 40.5 to 87.0% in liver, and from 52.5 to 74.5 in kidney. R.S.D.s were < 15.6% and < 10.6%, respectively. The detection and quantification limits were 0.05 and 0.5 mg kg(-1), for all MCs. The method was applied to MCs detection in liver and kidney of rat previously injected i.p. with MC-LR. Results showed the presence of MC-LR in the liver of the animals injected with the highest dose.     

Development and validation of a multi-residue method for the determination of pesticides in processed fruits and vegetables using liquid chromatography–electrospray ionization tandem mass spectrometry

A sensitive multi-residue analytical method, utilizing ethyl acetate extraction and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS), has been developed and validated for simultaneous determination of 28 pesticides of different chemical classes (polar organophosphates, carbamates, strobilurines, neonicotinoids, amides, pyrimidines, benzimidazoles, imidazoles and triazoles), and their transformation products, in processed fruit and vegetables. Two precursor-product ion transitions were monitored for each pesticide in selected reaction monitoring (SRM) mode. Linearity (r (2) > or = 0.99) was good over the concentration range 0.5 to 100 microg L(-1) for all the pesticides, and instrumental detection limits ranged from 0.1 to 1 microg L(-1). Mean recovery for fruit and vegetables spiked at 0.010 mg kg(-1) ranged from 65 to 94.4%, and relative standard deviations ranged from 9.0 to 20.0%. When the amount spiked was 0.050 mg kg(-1) recoveries ranged from 72.5 to 90% and relative standard deviations were from 6.1 to 19.0%. Method detection limits were from 0.002 to 0.007 mg kg(-1) for the different food matrices studied. The method was used to monitor pesticide residues in a wide variety of fruits and vegetables.     

Determination of phenolic compounds in wines by novel matrix solid-phase dispersion extraction and gas chromatography/mass spectrometry

A novel matrix solid-phase dispersion (MSPD) extraction method was developed to extract simultaneously 23 phenolic compounds from wine samples prior to determination by gas chromatography with mass spectrometric detection in the selected ion monitoring mode. Different parameters of the MSPD technique such as dispersant solid-phase, eluting solvent, and sample ionic strength and pH were optimized. The optimized MSPD procedure requires a small volume of wine (1 mL), commercial silica gel (1.5 g) as dispersant solid-phase and a small volume of ethyl acetate (5 mL) as eluting solvent. Under these conditions, the extraction of the studied compounds was almost complete (mean values of recoveries between 87 and 109%) in a short time (15 min). Moreover, satisfactory standard deviations of repeatability (RSD<9% in most cases), linear regression coefficients (r(2)>0.993) and detection limits (<8 microg/L) confirm the usefulness of the methodology for routine monitoring of the concentration of individual phenolic antioxidants in wines. Application was illustrated by analysis of different wine samples.     

Multiresidue analysis of pesticides in animal liver by gas chromatography using triple quadrupole tandem mass spectrometry

Two methods for extracting organochlorine (OCs) and organophosphorus (OPs) pesticides from animal liver have been developed. The determination was carried out by gas chromatography with electron impact ionization tandem mass spectrometry (GC-(EI-)MS/MS) using a triple quadrupole (QqQ) analyzer. First, a liquid-solid extraction performed with a high-speed homogenizer (Polytron) using ethyl acetate as solvent, and a subsequent clean-up by gel permeation chromatography (GPC) was applied, determining 34 pesticides. Secondly, a matrix solid phase dispersion (MSPD) extraction with octadecylsilyl (C(18)) sorbent combined with a Florisil clean-up and ethyl acetate elution was performed, analyzing 25 compounds. These methodologies have been tested and compared in the sample pre-treatment due to the fatty nature of the matrix. The GPC method was finally selected and validated, yielding recoveries in the range 70-115%, with precision values expressed as relative standard deviation (RSD) lower or equal to 20%, at the spiking levels of 25 and 50 microg kg(-1), and limits of quantification (LOQs) lower than the maximum residue levels (MRLs) set by the European Union in animal products, except for isofenphos. Linearity was also studied ranging between 5 and 300 microg kg(-1) for most of pesticides. This method was applied to the analysis of real liver samples of chicken, pork and lamb.     

Determination of thiabendazole in orange juice and rind by liquid chromatography with fluorescence detection and confirmation by gas chromatography/mass spectrometry after extraction by matrix solid-phase dispersion

A method was developed for the determination of thiabendazole (TBZ) in orange juice and rind based on matrix solid-phase dispersion (MSPD). TBZ was extracted with ethyl acetate and the extract was subsequently cleaned up on a solid-phase extraction column. Fungicide residues were determined by liquid chromatography with fluorescence detection. Recoveries through the method ranged from 87 to 97% with relative standard deviations < or = 11%. The detection and quantitation limits were 0.15 and 0.50 microg/kg, respectively. The confirmation of TBZ residues in positive samples was performed by solid-phase microextraction followed by gas chromatography with mass spectrometric detection using selected ion monitoring. The developed method was applied to determine TBZ levels in commercial orange juices and in juice and rind of fresh oranges. The influence of storage and washing of fruits on TBZ residues was also studied.     

Matrix solid-phase dispersion extraction of carbadox and olaquindox in feed followed by hydrophilic interaction ultra-high-pressure liquid chromatographic analysis

A new method involving matrix solid-phase dispersion (MSPD) extraction and hydrophilic interaction ultra-high-pressure liquid chromatography (HILIC-UHPLC) with photodiode array detection was developed for the determination of carbadox and olaquindox in feed. Separation of carbadox and olaquindox was achieved within 1 min on the 1.7 microm Acquity UPLC BEH HILIC column by using isocratic elution with a mobile phase consisting of 10 mmol L(-1) ammonium acetate in acetonitrile-water (95:5, v/v) at a flow rate of 0.5 mL min(-1). Optimization of MSPD extraction parameters, such as type of solid sorbent and elution solvent were carried out. Optimal conditions selected for MSPD extraction were: 0.25 g of feed sample, 0.5 g of octadecylsilica as solid sorbent and 10 mL of acetonitrile-methanol (8:2, v/v) as eluting solvent. Both analytes provided average recoveries from spiked feed samples ranging from 89.1 to 98.4% with relative standard deviations less than 10%. Obtained performance characteristics are comparable to those achieved by liquid-liquid extraction-HPLC with the advantages of being simpler and significantly faster.     

Determination of organophosphorus pesticides in bovine tissue by an on-line coupled matrix solid-phase dispersion-solid phase extraction-high performance liquid chromatography with diode array detection method

A miniaturized method based on matrix solid-phase dispersion coupled to solid phase extraction and high performance liquid chromatography with diode array detection (MSPD-SPE-HPLC/DAD) was developed for the trace simultaneous determination of the following organophosphorus pesticides (OPPs) in bovine tissue: parathion-methyl, fenitrothion, parathion, chlorfenvinphos, diazinon, ethion, fenchlorphos, chlorpyrifos and carbophenothion. To perform the coupling between MSPD and SPE, 0.05 g of sample was dispersed with 0.2 g of C(18) silica sorbent and packed into a stainless steel cartridge containing 0.05 g of silica gel in the bottom. After a clean-up of high and medium polarity interferences with water and an acetonitrile:water mixture, the OPPs were desorbed from the MSPD cartridge with pure acetonitrile and directly transferred to a dynamic mixing chamber for dilution with water and preconcentration into an SPE 20 mm × 2.0 mm I.D. C(18) silica column. Subsequently, the OPPs were eluted on-line with the chromatographic mobile phase to the analytical column and the diode array detector for their separation and detection, respectively. The method was validated and yielded recovery values between 91% and 101% and precision values, expressed as relative standard deviations (RSD), which were less than or equal to 12%. Linearity was good and ranged from 0.5 to 10 μg g(-1), and the limits of detection of the OPPs were in the range of 0.04-0.25 μg g(-1). The method was satisfactorily applied to the analysis of real samples and is recommended for food control, research efforts when sample amounts are limited, and laboratories that have ordinary chromatographic instrumentation.     

Ultrasonic-assisted matrix solid-phase dispersion as an improved methodology for the determination of pesticides in fruits

An ultrasonic-assisted matrix solid-phase dispersion (UA-MSPD) method has been developed for extracting and cleaning-up 15 organophosphorus pesticides (OPPs) and 9 triazines in fruits. Using reversed phase octasilyl-derivatised silica (C8) as dispersant and ethyl acetate as extraction solvent, two sonication devices, an ultrasonic bath and a sonoreactor, were tested for speeding and increasing the efficiency of the MSPD process. A standard MSPD and a heating-assisted MSPD procedure were also done. Gas chromatography-mass spectrometry was used for analyte determination. 1-min sonication with the sonoreactor at 50% amplitude provided the best results, with reproducibilities below 15% for the pesticides studied. The general low detection limits (1-42microgkg(-1)) ensure proper determination at maximum allowed residue levels set in current legislations, except for dimethoate and disulfuton. The analytical performance of the method was evaluated for apples, pear and apricot, showing little or no matrix effect.     

Determination of imidacloprid, metalaxyl, myclobutanil, propham, and thiabendazole in fruits and vegetables by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry

Imidacloprid, metalaxyl, myclobutanil, propham, and thiabendazole have been simultaneously determined in strawberries, oranges, potatoes, pears, and melons by matrix solid-phase dispersion (MSPD) followed by liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS) in positive-ion mode. The samples were homogenized with C8 bonded silica as MSPD sorbent, placed in a glass column, and eluted with dichloromethane. Chromatographic separation of the compounds was achieved on a reversed-phase LC column using a methanol-ammonium formate (50 mmol L(-1)) gradient as a mobile phase. Samples were screened by monitoring the protonated molecular ion at m/z 256 for imidacloprid, 280 for metalaxyl, 289 for myclobutanil, and 202 for thiabendazole, and the main fragment at m/z 138 for propham. Positive samples were confirmed by multiple-ion monitoring. The repeatability (<20%) and recovery (>57%) of the method were good, and limits of detection (<0.05 mg kg(-1)) were adequate.     

Response surface optimization for determination of pesticide multiresidues by matrix solid-phase dispersion and gas chromatography

An optimized multiresidue analysis method based on matrix solid-phase dispersion (MSPD) and gas chromatography (GC) is proposed for the determination of organochlorines and pyrethroids in the tea samples. Response surface methodology (RSM) was used to optimize the extraction conditions of MSPD, such as the sorbent type, eluent composition, dichloromethane concentration and eluting volume. Desirability function approach was employed to optimize the pesticide recoveries and matrix cleanup. Compromising the recoveries and cleanup degree, MSPD was carried out with Florisil as the sorbent and n-hexane-dichloromethane (1:1, v/v) as the eluent. The pesticide recoveries in tea samples were better than 80% spiked in the concentration range of 0.01-0.05mg/kg and the relative standard deviations were lower than 7%. The quantification limits of the pesticides were in the range of 0.002-0.06mg/kg, which were lower than the maximum residue limits of the pesticides in tea samples established by the European Union.     

Routine application using single quadrupole liquid chromatography-mass spectrometry to pesticides analysis in citrus fruits

A rapid and sensitive liquid chromatography-electrospray ionization-mass spectrometry method has been developed for the routine analysis of buprofezin, bupirimate, hexaflumuron, tebufenpyrad, fluvalinate and pyriproxyfen in citrus fruits. Extracts were obtained by matrix solid-phase dispersion (MSPD) using C18 as dispersant and dichloromethane-methanol (80:20, v/v) as eluent. Matrix effects were tested for all matrices by addition of standard to sample blank extracts (samples containing no detectable residues). Mean recoveries obtained at fortification levels between 0.01 and 5 mg kg(-1) were 57-97% with relative standard deviations (RSDs) from 5 to 19%. The limits of quantification (LOQ) were in the range of 0.01-0.2 mg kg(-1) and lower than maximum residue limits (MRLs) established by the Spanish legislation. The MSPD was compared with conventional ethyl acetate extraction, showing equivalent recoveries and precision. Although the sample is more concentrated (5-fold) by solid-liquid extraction (SLE) with ethyl acetate than by MSPD, LOQs obtained by both techniques, were almost equal, because MSPD reduces matrix effects, baseline noise, and interfering peaks from the matrix. The proposed method has been applied to the determination of selected pesticides in real samples. Liquid chromatography-tandem mass spectrometry (LC-MS-MS) with quadrupole ion trap (QIT) and triple quadrupole (TQ) have been used as confirmatory tool for positive samples according to a recent No. SANCO/10476/2003 European Union Guideline.     

Automated extraction and preconcentration of multiresidue of pesticides on a micro-solid-phase extraction system based on polypyrrole as sorbent and off-line monitoring by gas chromatography-flame ionization detection

A simple and efficient automated method for extraction and preconcentration of some organophosphorus and organochlorine pesticides from drinking and agriculture waters has been developed. In this work, a pyrrole-based polymer was synthesized and applied as an efficient sorbent for micro-solid-phase extraction (mu-SPE). Polypyrrole (PPY) was synthesized by chemical oxidation of the monomer in nonaqueous solution. The mu-SPE of selected pesticides, from aqueous samples was performed using 20mg of PPY. After extraction, analytes were desorbed in ethyl acetate and analyzed using gas chromatography-flame ionization detection. The acid-base and other physico-chemical interactions with the analytes facilitated the adsorption of analytes, with good selectivity and reproducibility. Under the optimized extraction conditions, the method showed good linearity in the range of 0.001-0.3 microg ml(-1) of water sample (phosphate buffer 1.0 x 10(-3) M, pH 8.5), RSD=1.5-4.2% (n=4), and limits of detection 200-600 pg ml(-1). Each mu-SPE cartridge could be used for up to 20 extractions.     

Rapid method for the determination of polycyclic aromatic hydrocarbons in agricultural soils by sonication-assisted extraction in small columns

A rapid method has been developed for the determination of 16 polycyclic aromatic hydrocarbons (PAHs) in soil based on their sonication-assisted extraction in small columns (SAESC) with a low volume of ethyl acetate and subsequent quantitation and identification by GC with electron impact mass spectrometric detection in the SIM mode (GC-MS-SIM). Spiked blank soil extracts were used as standards to counteract the matrix effect observed in the chromatographic determination. PAHs were confirmed at trace level by their retention times, their qualifier and target ions, and their qualifier/target abundance ratios. Recovery studies were performed at 0.5, 1.0, 5.0, and 10 microg/kg fortification levels for each PAH, and the recoveries obtained ranged from 91.2 to 99.8% with RSDs between 0.4 and 9.3%. The detection limits of the method ranged from 0.03 to 0.3 microg/kg for the different PAHs studied. The developed method is linear over the range assayed, 1-100 microg/L with determination coefficients higher than 0.996. PAH levels were determined using this method in soil samples taken from different agricultural areas of Spain. In general, PAH concentrations were low and the most frequently occurring PAHs were naphthalene, pyrene, phenanthrene, and fluoranthene.