Search

The QuEChERSER mega-method has recently been introduced to quantify and identify a wide range of chemical residues (pesticides, veterinary drugs, environmental contaminants, among others) in nearly all types of foods. The approach calls for taking a small amount of the initial extract to cover analytes amenable to liquid chromatography, and the remainder is salted out for analysis by gas chromatography (GC), both with mass spectrometry (MS) based detection. In the case of GC-MS(/MS), the extract undergoes automated robotic mini-cartridge solid-phase extraction (SPE) cleanup in a technique known as µSPE or instrument-top sample preparation (ITSP). In 2022, a septumless mini-cartridge for µSPE was introduced to improve upon the ITSP design. The new design houses a bed of 20 mg anhydrous MgSO 4 , 12 mg each of C18 and primary secondary amine sorbents, and 1 mg of graphitized carbon black, the latter substituting for CarbonX used in the ITSP product. The septumless µSPE mini-cartridge employs a different gripping mechanism with the syringe needle that allows leak-free operation at higher flow rates (e.g. 10 µL/s), whereas the ITSP design is limited to 2 µL/s. Based on cleanup and analyte elution profiles, the extract load volume and flow rate was increased in µSPE for QuEChERSER from 300 µL at 2 µL/s to 500 µL at 5 µL/s, which improved accuracy of results, sped the cleanup step, and obviated the need for micro-vial inserts in the receiving vials. The new design also reduced both the amount and consistency of dead (void) volumes in the mini-cartridges from 83 ± 14 µL to 52 ± 7 µL for 200-600 µL load volumes. Normalization of peak areas to internal standards led to recoveries between 80 and 120% with typical RSDs <5% in low-pressure GC-MS/MS of 227-242 out of 252 pesticides, polychlorinated biphenyls, polybrominated diphenyl ethers, and polycyclic aromatic hydrocarbons in hemp powder, spinach, whole milk, egg, avocado, and lamb meat., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Published by Elsevier B.V.)

Hemp has been an agricultural commodity for millennia, and it has been undergoing a resurgence in interest and production due to its high content of cannabinoids, protein, fiber and other ingredients. For legal possession and use throughout the USA, hemp and hemp products must have delta-9-tetrahydrocannabinol (THC) concentration < 0.3%. As with most crops, pesticides may be applied when farming hemp, which need to be monitored in food, feed, and medicinal products. The aim of this work was to evaluate and validate the recently developed "quick, easy, cheap, effective, rugged, safe, efficient, and robust" (QuEChERSER) sample preparation mega-method to determine pesticide residues in hemp plants, flowers, powders, oils, and pellets. High-throughput analysis of final extracts for 106 targeted pesticides and metabolites from North American monitoring lists entailed: 1) ultrahigh-performance liquid chromatography - tandem mass spectrometry (UHPLC-MS/MS) with column back-flushing, and 2) instrument-top sample preparation + low-pressure gas chromatography (ITSP+LPGC-MS/MS). In QuEChERSER, 2 g sample is extracted with 10 mL 4/1 (v/v) acetonitrile/water by mechanical shaking for 10 min, followed by 3 min centrifugation. For LC, 0.2 mL of extract is taken and solvent exchanged into initial mobile phase followed by 5 min ultra-centrifugation prior to the 10 min analysis. For GC-amenable pesticides, the remaining initial extract is partitioned with 4/1 (w/w) anh. MgSO 4 /NaCl, and 1 mL is taken for automated ITSP cleanup in parallel with 10 min LPGC analysis. In the former case, the UHPLC column is back-flushed with 1/1 (v/v) methanol/acetonitrile for 3 min between each injection to keep the system clean and avoid ghost peaks. Multi-level, multi-day validation results achieved 70-120% recoveries with RSDs < 20% for more than 80% of the analytes in hemp protein powder, oil, pellets, and fresh plant (dried hemp plant and flower were too complex). Limits of quantification (LOQs) were < 10 ng/g were achieved for nearly all pesticides, yielding 2.8% false negatives among >13,000 analyte results in the spiked samples. The QuEChERSER method was demonstrated to meet the challenge for several complex hemp matrices., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier B.V.)

Despite nearly 80 years of advancements in gas chromatography (GC), indirect chemical matrix effects (MEs), known as the matrix-induced response enhancement effect, still occur to cause a high bias in the GC analysis of susceptible analytes, unless precautions are taken. Matrix-matched calibration is one common option used in GC to compensate for the MEs, but this approach is usually inconvenient, imprecise, and inefficient. Other options, such as the method of standard additions, surface deactivation techniques, chemical derivatizations, priming the GC, and/or use of internal standards, also have flaws in practice. When methods are accommodating, the use of analyte protectants (APs) can provide the best practical solution to not only overcome MEs, but also to maximize analyte signal by increasing chromatographic and detection efficiencies for the analytes. APs address the source of MEs in every injection by filling active sites in the GC inlet, column, and detector, particularly in GC-MS, rather than the analytes that would otherwise undergo degradation, peak tailing, and/or diminished response due to interactions with the active sites. The addition of an adequate amount of APs (e.g. sugar derivatives) to all calibration standards and final extracts alike often leads to lower detection limits, better accuracy, narrower peaks, and greater robustness than the other options to compensate for MEs in GC. This article consists of a critical review of the scientific literature, proposal of mechanisms and theory, and re-evaluation studies involving APs for the first time in GC-orbitrap and GC-MS/MS with a high-efficiency ion source design. The findings showed that 1 µg each of co-injected shikimic acid and sorbitol in the former case, and 1 µg shikimic acid alone in the latter case, led to high quality results in multi-residue analysis of pesticides and environmental contaminants., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Conventional gas chromatography - mass spectrometry (GC-MS) takes 20-40 min per sample, which is undesirably slow in any application if speed can be increased while still meeting analytical needs. In this study, we achieved reasonably good separations with full analysis cycle times of less than 1 min by combining for the first time low-pressure (LP) GC-MS with low thermal mass (LTM) resistive-heating for rapid temperature ramping and cooling of the capillary column. The analytical column is threaded into the LTM thin-walled metal tubing in an instrumental device known as "LTM Fast GC" that is mounted at the top of the gas chromatograph in a detector port. The column inlet and outlet are connected to the GC injector and MS transfer line as usual. For LPGC-MS, a 40 cm, 0.1 mm. i.d. uncoated flow restrictor capillary connected at the injector is coupled with a 2.6 m, 0.25 mm i.d., 0.25 µm film thickness analytical column leading to the MS. Thus, the inlet operates at normal GC pressures, but the analytical column is under vacuum, which increases the optimal helium carrier gas flow velocity thereby increasing speed of full range separations while maintaining acceptable quality of chromatography. This column configuration in LTM-LPGC-MS trades a 64-fold gain in speed of analysis vs. standard GC-MS for a 4-fold loss in chromatographic peak capacity, thereby converting analysis time from minutes into seconds in common applications. For example, jet fuel containing fatty acid methyl esters (akin to biofuel) was separated in 25 s with <1 min full analysis cycle time. An EPA Method 8270 mixture of 76 analytes was also analyzed in <1 min full cycle time by LTM-LPGC-MS. Other examples include very fast analysis of heroin in a street drug powder and elucidation of a new organic synthetic compound. In this report, we describe and discuss the several advantageous and practical features of LTM-LPGC-MS, as well as its trade-offs., (Copyright © 2019. Published by Elsevier B.V.)

This study demonstrates the application of a novel lipid removal product to the residue analysis of 65 pesticides and 52 environmental contaminants in kale, pork, salmon, and avocado by fast, low pressure gas chromatography - tandem mass spectrometry (LPGC-MS/MS). Sample preparation involves QuEChERS extraction followed by use of EMR-Lipid ("enhanced matrix removal of lipids") and an additional salting out step for cleanup. The optimal amount of EMR-Lipid was determined to be 500mg for 2.5mL extracts for most of the analytes. The co-extractive removal efficiency by the EMR-Lipid cleanup step was 83-98% for fatty samples and 79% for kale, including 76% removal of chlorophyll. Matrix effects were typically less than ±20%, in part because analyte protectants were used in the LPGC-MS/MS analysis. The recoveries of polycyclic aromatic hydrocarbons and diverse pesticides were mostly 70-120%, whereas recoveries of nonpolar polybrominated diphenyl ethers and polychlorinated biphenyls were mostly lower than 70% through the cleanup procedure. With the use of internal standards, method validation results showed that 76-85 of the 117 analytes achieved satisfactory results (recoveries of 70-120% and RSD≤20%) in pork, avocado, and kale, while 53 analytes had satisfactory results in salmon. Detection limits were 5-10ng/g for all but a few analytes. EMR-Lipid is a new sample preparation tool that serves as another useful option for cleanup in multiresidue analysis, particularly of fatty foods., (Published by Elsevier B.V.)

The concept of low pressure (LP) vacuum outlet gas chromatography (GC) was introduced more than 50 years ago, but it was not until the 2000s that its theoretical applicability to fast analysis of GC-amenable chemicals was realized. In practice, LPGC is implemented by placing the outlet of a short, wide (typically 10-15 m, 0.53 mm inner diameter) analytical column under vacuum conditions, which speeds the separation by reducing viscosity of the carrier gas, thereby leading to a higher optimal flow rate for the most separation efficiency. To keep the inlet at normal operating pressures, the analytical column is commonly coupled to a short, narrow uncoated restriction capillary that also acts as a guard column. The faster separations in LPGC usually result in worse separation efficiency relative to conventional GC, but selective detection usually overcomes this drawback. Mass spectrometry (MS) provides highly selective and sensitive universal detection, and nearly all GC-MS instruments provide vacuum outlet conditions for implementation of LPGC-MS(/MS) without need for adaptations. In addition to higher sample throughput, LPGC provides other benefits, including lower detection limits, less chance of analyte degradation, reduced peak tailing, increased sample loadability, and more ruggedness without overly narrow peaks that would necessitate excessively fast data acquisition rates. This critical review summarizes recent developments in the application of LPGC with MS and other detectors in the analysis of pesticides, environmental contaminants, explosives, phytosterols, and other semi-volatile compounds., (Published by Elsevier B.V.)

The "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) approach to sample preparation is widely applied in pesticide residue analysis, but the use of magnesium sulfate and other nonvolatile compounds for salting out in the method is not ideal for mass spectrometry. In this study, we developed and evaluated three new different versions of the QuEChERS method using more volatile salts (ammonium chloride and ammonium formate and acetate buffers) to induce phase separation and extraction of 43 representative pesticide analytes of different classes. Fast low-pressure gas chromatography tandem mass spectrometry (LPGC-MS/MS) and liquid chromatography (LC)-MS/MS were used for analysis. The QuEChERS AOAC Official Method 2007.01 was also tested for comparison purposes. Of the studied methods, formate buffering using 7.5g of ammonium formate and 15mL of 5% (v/v) formic acid in acetonitrile for the extraction of 15g of sample (5g for wheat grain) provided the best performance and practical considerations. Method validation was carried out with and without the use of dispersive solid-phase extraction for cleanup, and no significant differences were observed for the majority of pesticides. The method was demonstrated in quantitative analysis for GC- and LC-amenable pesticides in 4 representative food matrices (apple, lemon, lettuce, and wheat grain). With the typical exceptions of certain pH-dependent and labile pesticides, 90-110% recoveries and <10% RSD were obtained. Detection limits were mostly <5ng/g, which met the general need to determine pesticide concentrations as low as 10ng/g for monitoring purposes in food applications., (Published by Elsevier B.V.)

A new method of sample preparation was developed and is reported for the first time. The approach combines in-vial filtration with dispersive solid-phase extraction (d-SPE) in a fast and convenient cleanup of QuEChERS (quick, easy, cheap, effective, rugged, and safe) extracts. The method was applied to simultaneous analysis of 42 diverse pesticides and 17 environmental contaminants, including polycyclic aromatic hydrocarbons, polychlorinated biphenyls (PCBs), and flame retardants, in shrimp as the sample matrix. Final extracts were analyzed by both low-pressure gas chromatography - triple quadrupole tandem mass spectrometry (LPGC-MS/MS), and high-performance liquid chromatography - triple quadrupole tandem mass spectrometry (HPLC-MS/MS) to provide a wide scope of analysis for targeted analytes. During method development, several different commercial sorbents for d-SPE were investigated and compared with respect to analyte recoveries. The method was validated at 10, 50, and 100 ng g(-1) spiking levels (10-fold lower for PCBs), and the results for nearly all analytes were between 70 and 115% recoveries with ≤17% relative standard deviations. The method was shown to be simple, fast, and effective for multi-application analysis of chemical residues in the representative food and environmental marker matrix., (Copyright © 2014. Published by Elsevier B.V.)

A high-throughput qualitative screening and identification method for 9 aminoglycosides of regulatory interest has been developed, validated, and implemented for bovine kidney, liver, and muscle tissues. The method involves extraction at previously validated conditions, cleanup using disposable pipette extraction, and analysis by a 3 min ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method. The drug analytes include neomycin, streptomycin, dihydrosptreptomycin, and spectinomycin, which have residue tolerances in bovine in the US, and kanamicin, gentamicin, apramycin, amikacin, and hygromycin, which do not have US tolerances established in bovine tissues. Tobramycin was used as an internal standard. An additional drug, paromomycin also was validated in the method, but it was dropped during implementation due to conversion of neomycin into paromomycin. Proposed fragmentation patterns for the monitored ions of each analyte were elucidated with the aid of high resolution MS using a quadrupole-time-of-flight instrument. Recoveries from spiking experiments at regulatory levels of concern showed that all analytes averaged 70-120% recoveries in all tissues, except hygromycin averaged 61% recovery. Lowest calibrated levels were as low as 0.005 μg/g in matrix extracts, which approximately corresponded to the limit of detection for screening purposes. Drug identifications at levels <0.05 μg/g were made in spiked and/or real samples for all analytes and tissues tested. Analyses of 60 samples from 20 slaughtered cattle previously screened positive for aminoglycosides showed that this method worked well in practice. The UHPLC-MS/MS method has several advantages compared to the previous microbial inhibition screening assay, especially for distinguishing individual drugs from a mixture and improving identification of gentamicin in tissue samples., (Published by Elsevier B.V.)

In the field of food contaminant analysis, the most significant development of recent years has been the integration of ultra-high pressure liquid chromatography (UHPLC), coupled to tandem quadrupole mass spectrometry (MS/MS), into analytical applications. In this review, we describe the emergence of UHPLC through technological advances. The implications of this new chromatographic technology for MS detection are discussed, as well as some of the remaining challenges in exploiting it for chemical residue applications. Finally, a comprehensive overview of published applications of UHPLC-MS in food contaminant analysis is presented, with a particular focus on veterinary drug residues., (Copyright © 2013 Elsevier B.V. All rights reserved.)

A multi-class, multi-residue method for the analysis of 13 novel flame retardants, 18 representative pesticides, 14 polychlorinated biphenyl (PCB) congeners, 16 polycyclic aromatic hydrocarbons (PAHs), and 7 polybrominated diphenyl ether (PBDE) congeners in catfish muscle was developed and evaluated using fast low pressure gas chromatography triple quadrupole tandem mass spectrometry (LP-GC/MS-MS). The method was based on a QuEChERS (quick, easy, cheap, effective, rugged, safe) extraction with acetonitrile and dispersive solid-phase extraction (d-SPE) clean-up with zirconium-based sorbent prior to LP-GC/MS-MS analysis. The developed method was evaluated at 4 spiking levels and further validated by analysis of NIST Standard Reference Materials (SRMs) 1974B and 1947. Sample preparation for a batch of 10 homogenized samples took about 1h/analyst, and LP-GC/MS-MS analysis provided fast separation of multiple analytes within 9min achieving high throughput. With the use of isotopically labeled internal standards, recoveries of all but one analyte were between 70 and 120% with relative standard deviations less than 20% (n=5). The measured values for both SRMs agreed with certified/reference values (72-119% accuracy) for the majority of analytes. The detection limits were 0.1-0.5ng g(-1) for PCBs, 0.5-10ng g(-1) for PBDEs, 0.5-5ng g(-1) for select pesticides and PAHs and 1-10ng g(-1) for flame retardants. The developed method was successfully applied for analysis of catfish samples from the market., (Published by Elsevier B.V.)

Gas and liquid chromatography (GC and LC) coupled to mass spectrometry (MS) serve as the most powerful analytical tools commonly used to monitor pesticide residues in food, among other applications. However, both GC-MS and LC-MS are susceptible to matrix effects which can adversely affect quantification depending on the analyte, matrix, sample preparation, instrumentation, and operating conditions. Among the approaches that reduce matrix effects, the most common in pesticide residue applications is matrix-matched calibration because it is relatively inexpensive and simple. Also, it has been shown to work well during method validation when fortified samples are exactly matched with samples used for calibration. However, the quality of matrix-matched results in real-world analyses depends on the consistency of matrix effects among diverse samples. In this study, the variability of matrix effects was measured for 38 representative pesticides in 20 samples each (including different varieties) of rice, orange, apple, and spinach extracted using the "quick, easy, cheap, effective, rugged, and safe" (QuEChERS) method for analysis by LC-MS/MS and low-pressure GC-MS. Using LC-MS/MS, only oranges gave >20% matrix effects for a few pesticides. GC-MS exhibited larger matrix effects, but as in LC-MS/MS, the differences were reasonably consistent among the 20 samples tested. Main conclusions of this study are that for the conditions utilized: (1) matrix-matching was not needed for most pesticides in the simpler food matrices; and (2) for the more complex orange matrix, acceptably accurate quantitative results were achieved by using matrix-matching even with a different sample of the same type. However, full confidence cannot be extended to matrix-matched results, and for consequential applications such as regulatory enforcement, confirmatory analyses using alternate quantitative determinations should also be conducted., (Published by Elsevier B.V.)

In this study, optimization, extension, and validation of a streamlined, qualitative and quantitative multiclass, multiresidue method was conducted to monitor >100 veterinary drug residues in meat using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Optimization centered on extensive ruggedness evaluation of the method. Various clean-up sorbents were tested and the amount of co-extractives were weighed, matrix effects were measured using post-column infusion of representative analytes, the effect of extract dilution before injecting was studied, and analyte recoveries and reproducibilities were determined. In order to extend our previous method, more drug analytes were added that possessed a wider range of chemical properties, and a re-appraisal of different types of C18 in dispersive solid-phase extraction clean-up and mobile phases in UHPLC-MS/MS was done. Ultimately, end-capped C18 and post-column infusion of ammonium formate as an ionization enhancer for the late-eluting anthelmintics were found to give improved qualitative results for greater analytical scope. A multi-day, multi-analyst validation demonstrated that the final method is suitable for screening of 113 analytes, identifying 98 and quantifying (recoveries between 70-120% and RSD<25%) 87 out of the 127 tested drugs at or below US regulatory tolerance levels in bovine muscle., (Published by Elsevier B.V.)

We developed and evaluated a new method of low-pressure gas chromatography-tandem mass spectrometry (LP-GC/MS-MS) using a triple quadrupole instrument for fast analysis of 150 relevant pesticides in four representative fruits and vegetables. This LP-GC (vacuum outlet) approach entails coupling a 10 m, 0.53 mm i.d., 1 μm film analytical column between the MS transfer line and a 3 m, 0.15 mm i.d. capillary at the inlet. The MS creates a vacuum in the 10 m analytical column, which reduces the viscosity of the He carrier gas and thereby shifts the optimal flow rate to greater velocity. By taking advantage of the H(2)-like properties of He under vacuum, the short analytical column, a rapid oven temperature ramp rate, and the high selectivity and sensitivity of MS/MS, 150 pesticides were separated in <6.5 min. The 2.5 ms dwell time and 1 ms interscan delay of the MS/MS instrument were critical for achieving >8 data points across the 2-3 s wide peaks. To keep dwell and cycle times constant across all peaks, each segment consisted of 30 analytes (60 transitions). For assessment, we injected extracts of spiked broccoli, cantaloupe, lemon, and sweet potato from the updated QuEChERS sample preparation method. Average recoveries (n=72) were 70-120% for 144 of the pesticides, and reproducibilities were <20% RSD for all but 4 analytes. Also, detection limits were <5 ng/g for all but a few pesticides, depending on the matrix. In addition to high quality performance, the method gave excellent reliability and high sample throughput, including easy peak integration to obtain rapid results., (Published by Elsevier B.V.)

This paper describes a method for the detection and quantification of 38 residues of the most widely used anthelmintics (including 26 veterinary drugs belonging to the benzimidazole, macrocyclic lactone and flukicide classes) in bovine liver using two different protocols for MRL and non-MRL levels. A dual validation approach was adopted to reliably quantify anthelmintic residues over an extended concentration range (1-3000 μg kg(-1)). Sample extraction and purification was carried out using a modified QuEChERS method. A concentration step was included when analysing in the low μg kg(-1) range. Rapid analysis was carried out by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), which was capable of detecting residues to <2 μg kg(-1). The method has been single-laboratory validated according to the 2002/657/EC guidelines and met acceptability criteria in all but a few cases. The inclusion of 19 internal standards, including 14 isotopically labelled internal standards, improved accuracy, precision, decision limit (CCα) and detection capability (CCβ)., (Copyright © 2010 Elsevier B.V. All rights reserved.)

A higher monitoring rate is highly desirable in the labs, but this goal is typically limited by sample throughput. In this study, we sought to assess the real-world applicability of fast, low-pressure GC-time-of-flight MS (LP-GC/TOFMS) for the identification and quantification of 150 pesticides in tomato, strawberry, potato, orange, and lettuce samples. Buffered and unbuffered versions of QuEChERS (which stands for "quick, easy, cheap, effective, rugged, and safe") using dispersive solid-phase extraction (d-SPE) and disposable pipette extraction (DPX) for clean-up were compared for sample preparation. For clean-up of all sample types, a combination of 150 mg MgSO₄, 50mg primary secondary amine (PSA), 50 mg C₁₈, and 7.5 mg graphitized carbon black (GCB) per mL extract was used. No significant differences were observed in the results between the different sample preparation versions. QuEChERS took < 10 min per individual sample, or < 1 h for two chemists to prepare 32 pre-homogenized samples, and using LP-GC/TOFMS, < 10 min run time and < 15 min cycle time allowed > 32 injections in 8 h. Overall, > 126 analytes gave recoveries (3 spiking levels) in the range of 70-120% with < 20% RSD. The results indicate that LP-GC/TOFMS for GC-amenable analytes matches UHPLC-MS/MS in terms of sample throughput and turnaround time for their routine, concurrent use in the analysis of a wide range of analytes in QuEChERS extracts to achieve reliable quantification and identification of pesticide residues in foods., (Published by Elsevier B.V.)

A new UHPLC-MS/MS (ultra high performance liquid chromatography coupled to tandem mass spectrometry) method was developed and validated to detect 38 anthelmintic drug residues, consisting of benzimidazoles, avermectins and flukicides. A modified QuEChERS-type extraction method was developed with an added concentration step to detect most of the analytes at <1 microg kg(-1) levels in milk. Anthelmintic residues were extracted into acetonitrile using magnesium sulphate and sodium chloride to induce liquid-liquid partitioning followed by dispersive solid phase extraction for cleanup. The extract was concentrated into dimethyl sulphoxide, which was used as a keeper to ensure analytes remain in solution. Using rapid polarity switching in electrospray ionisation, a single injection was capable of detecting both positively and negatively charged ions in a 13 min run time. The method was validated at two levels: the unapproved use level and at the maximum residue level (MRL) according to Commission Decision (CD) 2002/657/EC criteria. The decision limit (CCalpha) of the method was in the range of 0.14-1.9 and 11-123 microg kg(-1) for drugs validated at unapproved and MRL levels, respectively. The performance of the method was successfully verified for benzimidazoles and levamisole by participating in a proficiency study.

This article describes the comparison of different versions of an easy, rapid and low-cost sample preparation approach for the determination of pesticide residues in fruits and vegetables by concurrent use of gas and liquid chromatography (GC and LC) coupled to mass spectrometry (MS) for detection. The sample preparation approach is known as QuEChERS, which stands for "quick, easy, cheap, effective, rugged and safe". The three compared versions were based on the original unbuffered method, which was first published in 2003, and two interlaboratory validated versions: AOAC Official Method 2007.01, which uses acetate buffering, and European Committee for Standardization (CEN) Standard Method EN 15662, which calls for citrate buffering. LC-MS/MS and GC-MS analyses using each method were tested from 50 to 1000ng/g in apple-blueberry sauce, peas and limes spiked with 32 representative pesticides. As expected, the results were excellent (overall average of 98% recoveries with 10% RSD) using all 3 versions, except the unbuffered method gave somewhat lower recoveries for the few pH-dependent pesticides. The different methods worked equally well for all matrices tested with equivalent amounts of matrix co-extractives measured, matrix effects on quantification and chemical noise from matrix in the chromatographic backgrounds. The acetate-buffered version gave higher and more consistent recoveries for pymetrozine than the other versions in all 3 matrices and for thiabendazole in limes. None of the versions consistently worked well for chlorothalonil, folpet or tolylfluanid in peas, but the acetate-buffered method gave better results for screening of those pesticides. Also, due to the recent shortage in acetonitrile (MeCN), ethyl acetate (EtOAc) was evaluated as a substitute solvent in the acetate-buffered QuEChERS version, but it generally led to less clean extracts and lower recoveries of pymetrozine, thiabendazole, acephate, methamidophos, omethoate and dimethoate. In summary, the acetate-buffered version of QuEChERS using MeCN exhibited advantages compared to the other tested methods in the study., (Published by Elsevier B.V.)

A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) multi-residue method for the simultaneous quantification and identification of 38 residues of the most widely used anthelmintic veterinary drugs (including benzimidazoles, macrocyclic lactones, and flukicides) in milk and liver has been developed and validated. For sample preparation, we used a simple modification of the QuEChERS method, which was initially developed for pesticide residue analysis. The method involved extracting sample (10 g) with acetonitrile (10 mL), followed by phase separation from water (salting out) with MgSO(4):NaCl (4:1, w/w). After centrifugation, an aliquot of the extract (1 mL) was purified by dispersive solid-phase extraction with MgSO(4) (150 mg) and C(18) (50mg), prior to LC-MS/MS analysis. Two injections of the same extract were required with the LC-MS/MS instrument to cover the 30 electrospray positive and 8 electrospray negative analytes. The limit of quantitation of the method was 5 microgkg(-1) for 37 analytes (and 10 microgkg(-1) for dichlorvos). The method was successfully validated according to the 2002/657/EC guidelines. Recovery of analytes was typically in the 70-120% range, with repeatabilities and reproducibilities typically <15% in milk and <20% in liver.

Rapid screening tests can be used as part of an efficient program designed to monitor veterinary drug residues in cattle. In this work, three rapid tests designed to screen samples for the presence of antibiotic residues, the Fast Antimicrobial Screen Test (FAST), Premi and Kidney Inhibition Swab (KIS) tests, were compared using beef kidney juice and serum samples. In order to provide a realistic assessment, potentially incurred samples of beef kidney juice and serum were obtained from 235 carcasses which had been retained by inspectors in a processing plant for further testing. In addition, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was conducted on these samples to identify what antibiotics were present, if any, and their levels. The comparison of the three rapid screening test results with those from LC-MS/MS analysis allowed for a more complete comparison of the relative sensitivity of these analytical methods, as well as valuable information on false positive and negative response rates.

An automated direct sample introduction technique coupled to comprehensive two-dimensional gas chromatography-time of flight mass spectrometry (DSI-GC x GC/TOF-MS) was applied for the development of a relatively fast and easy analytical screening method for 17 polychlorinated dibenzo-p-dioxins/dibenzofurans (PCDD/Fs) and 4 non-ortho polychlorinated biphenyls (PCBs) in fish oil. Comparison of instrumental performance between DSI-GC x GC/TOF-MS and the traditional gas chromatographic high resolution mass spectrometric (GC-HRMS) method showed good agreement of results for standard solutions analyzed in blind fashion. Relatively high tolerance of the DSI technique for lipids in the final extracts enabled a streamlined sample preparation procedure that only required gel permeation chromatography (GPC) and solid-phase extraction (SPE) cleanup with graphitized carbon black. The sample size for the method was 2g of cod liver oil, which achieved limits of quantitation (LOQs) of 0.019-7.8 pg/g toxic equivalent quotients for the individual PCDD/Fs. Lower detection limits can be achieved by using larger sample size and scaling up the sample preparation procedure, but this adds to the labor, time, solvent consumption, and expense of the approach. However, the streamlined method yielded 0.94 pg/g and 2.3 pg/g LOQs for 2,3,7,8-tetrachloro dibenzofuran (TCDF) and 3,3',4,4',5-pentachloro biphenyl (CB126), which were sufficiently low for regulatory monitoring of 2g samples. Therefore, instead of congener specific analysis, this streamlined analytical screening method for TCDF and CB126 has the potential to monitor fish oil contaminated with dioxin and dioxin-like PCBs at or above current food safety limits. Acceptable recoveries for nearly all analytes at three different spiking levels in fish oil samples were achieved with good repeatability.

A rapid method using programmed temperature vaporiser injection-low-pressure gas chromatography-high-resolution time-of-flight mass spectrometry (PTV-LP-GC-HR-TOF-MS) for the analysis of multiple pesticide residues in fruit-based baby food was developed. The fast and inexpensive buffered QuEChERS (quick, easy, cheap, effective, rugged, and safe) extraction method and "conventional" approach that employs ethyl acetate extraction followed by gel permeation chromatography (GPC) cleanup were employed for sample preparation. A PTV injector in solvent venting mode was used to reduce volume of acetonitrile and acetic acid (from the buffered QuEChERS extracts) that caused higher column bleed without their elimination. Otherwise, the time-to-digital converter would become saturated in HR-TOF-MS. For fast GC separation allowing analysis of 100 analytes within a 7 min runtime, both a high temperature programming rate and vacuum conditions in a megabore GC column were employed. The use of HR-TOF-MS allowed the unbiased identification and reliable quantification of target analytes through the application of a narrow mass window (0.02 Da) for extracting analyte ions and the availability of full spectral information even at very low levels. With only a few exceptions, the lowest calibration levels for the pesticides tested were

The 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are among the most toxic compounds known, and several sources of exposure to these chemicals should be monitored to protect human and environmental health. The current predominant method of analysis is too expensive and cumbersome, and comprehensive two-dimensional GC coupled to time-of-flight mass spectrometry (GC x GC--TOF-MS) has the potential to lower the costs and speed analysis of PCDD/Fs. In this study, GC x GC--TOF parameters were evaluated and optimized to yield complete separation of the 17 most important PCDD/F congeners from polychlorinated biphenyls (PCBs) interferences, and to attain the lowest detection limits. The optimization study entailed evaluation of oven temperature programs, column flow rates, ion source temperatures, electron ionization energy, data acquisition rate, and various GC x GC parameters, including modulation period, modulator temperature offset and hot pulse duration. After optimization, all 17 PCDD/Fs were separated in <60 min, and in particular, the critical pair of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and pentachlorobiphenyl congener CB126 did not co-elute chromatographically. Accurate identification and determination of all analytes could be made using their deconvoluted full mass spectra. In GC x GC, the modulation period and start time were the most important factors that affected sensitivity and selectivity for the analysis of the PCDD/Fs. The modulation period should be < or = 4s to preserve separations achieved in one-dimensional GC, and the modulation start time was important to achieve one large slice and two smaller symmetrical slices of TCDD to maximize its detection sensitivity. After optimization, the method could identify 0.25 pg of TCDD with standard injection from its full mass spectrum.

Low-pressure gas chromatography-mass spectrometry (LP-GC-MS) using a quadrupole MS instrument was further optimized and evaluated for the fast analysis of multiple pesticide residues in food crops. Performance of two different LP-GC-MS column configurations was compared in various experiments, including ruggedness tests with repeated injections of pesticides in matrix extracts. The tested column configurations employed the same 3 m x 0.15 mm i.d. restriction capillary at the inlet end, but different analytical columns attached to the vacuum: (A) a 10 m x 0.53 mm i.d., 1 microm film thickness RTX-5 Sil MS column; and (B) a 10 m x 0.25 mm i.d., 0.25 microm film thickness DB-5MS column. Under the optimized conditions (compromise between speed and sensitivity), the narrower analytical column with a thinner film provided slightly (<1.1-fold) faster analysis of <5.5 min separation times and somewhat greater separation efficiency. However, lower detection limits for most of the tested pesticides in real extracts were achieved using the mega-bore configuration, which also provided significantly greater ruggedness of the analysis (long-term repeatability of analyte peak intensities, shapes, and retention times). Additionally, the effect of the increasing injection volume (1-5 microl) on analyte signal-to-noise ratios was evaluated. For the majority of the tested analyte-matrix combinations, the increase in sensitivity caused by a larger injection did not translate in the same gain in analyte detectability. Considering the costs and benefits, the injection volume of 2-3 microl was optimal for detectability of the majority of 57 selected pesticides in apple, carrot, lettuce, and wheat extracts.

An europium-sensitized time-resolved luminescence (TRL) method was developed to determine oxytetracycline (OTC) in cultivated catfish muscle. Extraction of OTC from fish muscle was performed with pH 4.0 ethylenediaminetetraacetic acid (EDTA)-McIlvaine buffer and clean up with hydrophilic-lipophilic balanced copolymer solid phase extraction (SPE) cartridges. The eluate was used without further concentration for TRL measurement in pH 9.0 micellar tris(hydroxylmethyl)aminomethane (TRIS) buffer. Cetyltrimethylammonium chloride (CTACl) was used as surfactant and EDTA as a co-ligand. The excitation and emission wavelengths were set at 388 and 615nm, respectively. The linear dynamic range was 0-1000ngg(-1) (R(2)=0.9995). The recovery was 92-112% in the fortification range of 50-200ngg(-1) and the limits of detection (LOD) ranged from 3 to 7ngg(-1). Incurred catfish samples were used to demonstrate the performance of the method around 100ngg(-1), the European Union maximum residue level.

In this study, we evaluated the suitability of six common organic solvents for gas chromatographic (GC) analysis of pesticides. Three of these, acetone, acetonitrile (MeCN) and ethyl acetate (EtAc), represent extraction solvents commonly used in multiresidue methods for determination of pesticides in produce. The other three, isooctane, hexane and toluene, often serve as exchange solvents before a GC analysis. An ideal solvent for GC analysis of multiclass pesticide residues should be compatible with: the analytes, sample preparation, and GC analysis. This study addresses each aspect with emphasis placed on stability of selected pesticides in the given solvents. In this respect, the exchange solvents proved to be superior to the more polar extraction solvents. Degradation of N-trihalomethylthio fungicides (e.g., captan, folpet, dichlofluanid) in MeCN was observed only in certain lots of the tested MeCN, but even if it occurred, the stability of these analytes as well as that of dicofol and chlorothalonil was dramatically improved by the addition of 0.1% (v/v) acetic acid. Dicofol and chlorothalonil were also unstable in acetone, and pesticides with a thioether group (e.g., fenthion, disulfoton) degraded in the tested EtAc. Formation of isomers of certain pyrethroids (deltamethrin, lambda-cyhalothrin) was recorded in the chromatograms from MeCN and acetone solutions, but this effect more likely occurred during the GC injection than in solution. For several reasons, MeCN was found to be the most suitable solvent for extraction of a wide polarity range of pesticide residues from produce. After acidification, the stability of problematic pesticides in MeCN is acceptable, and MeCN can also serve as a medium for GC injection; therefore solvent exchange is generally not required before GC analysis. If sensitivity is an issue in splitless injection, then toluene was demonstrated to be the best exchange solvent due to its miscibility with MeCN and stronger responses of relatively more polar pesticides (e.g., acephate, methamidophos) as compared to hexane and isooctane.

A common problem in gas chromatography (GC) applications is the analyte losses and/or peak tailing due to undesired interactions with active sites in the inlet and column. Analytes that give poor peak shapes or degrade have higher detection limits, are more difficult to identify and integrate, and are more prone to interferences than stable analytes that give narrow peaks. For susceptible analytes, significant peak quality improvements are obtained when matrix components are present because they fill active sites, thus reducing analyte interactions. This phenomenon is called "matrix-induced chromatographic response enhancement." Several approaches have been proposed to minimize peak distortion phenomena and compensate for matrix-induced effects, which is especially important for accurate quantitation, but each approach has serious limitations for routine multi-pesticide analysis. In this study, we demonstrate the feasibility of using "analyte protectants" to provide a more convenient and effective solution to the problem than other approaches developed thus far. The protecting agents are added to extracts and matrix-free standards alike to provide the chromatographic enhancement effect even for the most susceptible analytes in a very dirty GC system. In this study, we evaluated 93 different compounds to find the most suitable ones for improving chromatographic quality of the signal. Because hydrogen bonding has been shown to be an important factor in analyte interactions with active sites, we mainly focused on additives with strong hydrogen bonding capabilities. Dramatic peak enhancements were achieved using compounds containing multiple hydroxy groups, such as sugars and sugar derivatives, and gulonolactone appears to be the most effective protecting agent for the most pesticides that we tested. The benefits of using analyte protectants versus alternative procedures for overcoming matrix-induced effects in quantitation include: (a) simpler procedure; (b) easier integration of peaks; (c) lower detection limits; (d) better quantitation; (e) less maintenance of the GC inlet; and (e) lower cost. However, long-term influences on the performance of the chromatographic system have yet to be established.

Fast gas chromatography-mass spectrometry (GC-MS) has the potential to be a powerful tool in routine analytical laboratories by increasing sample throughput and improving laboratory efficiency. However, this potential has rarely been met in practice because other laboratory operations and sample preparation typically limit sample throughput, not the GC-MS analysis. The intent of this article is to critically review current approaches to fast analysis using GC-MS and to discuss practical considerations in addressing their advantages and disadvantages to meet particular application needs. The practical ways to speed the analytical process in GC and MS individually and in combination are presented, and the trade-offs and compromises in terms of sensitivity and/or selectivity are discussed. Also, the five main current approaches to fast GC-MS are described, which involve the use of: (1) short, microbore capillary GC columns; (2) fast temperature programming; (3) low-pressure GC-MS; (4) supersonic molecular beam for MS at high GC carrier gas flow; and (5) pressure-tunable GC-GC. Aspects of the different fast GC-MS approaches can be combined in some cases, and different mass analyzers may be used depending on the analytical needs. Thus, the capabilities and costs of quadrupole, ion trap, time-of-flight, and magnetic sector instruments are discussed with emphasis placed on speed. Furthermore, applications of fast GC-MS that appear in the literature are compiled and reviewed. At this time, the future usefulness of fast GC-MS depends to some extent upon improvement of existing approaches and commercialization of interesting new techniques, but moreover, a greater emphasis is needed to streamline overall laboratory operations and sample preparation procedures if fast GC-MS is to become implemented in routine applications.

We developed a new instrumental approach, termed Supersonic GC-MS, which achieves fast, sensitive, confirmatory and quantitative analysis of a broad range of pesticides in complex agricultural matrices. Our Supersonic GC-MS system is a modification of a bench-top Agilent 6890 GC+5972 MSD with a supersonic molecular beam (SMB) interface and fly-through EI ion source. One of the main advantages of Supersonic GC-MS is an enhanced molecular ion (M+) in the resulting mass spectra. For example, the M+ was observed in all 88 pesticides that we studied using the Supersonic GC-MS whereas only 36 of 63 (57%) pesticides that we investigated in standard GC-MS exhibited a M+. We also found that the degree of matrix interference is exponentially reduced with the fragment mass by about 20-fold per 100 amu increasing mass. The enhancement of the M+ combined with the reduction in matrix background noise permit rapid full scan analysis of a potentially unlimited number of pesticides, unlike selected ion monitoring or MS-MS in which specific conditions are required in segments for targeted pesticides. Furthermore, unlike the case with chemical ionization, EI-SMB-MS spectra still give accurate identification of compounds using common mass spectral libraries. In practice,we found thatlibraries favor mass spectra in which the M+ appears, thus Supersonic GC-MS produced better spectra for compound identification than standard GC-MS. To achieve even lower identification limits, the M+ plus a second major ion (still using full scan data) gives higher signal-to-chemical noise ratios than the traditional 3-ion approach. The replacement of two low-mass ions with the M+ (supersonic two-ions method) results in a significant reduction of matrix interference by a factor of up to 90. Another main advantage of Supersonic GC-MS is its exceptional suitability for fast GC-MS with high carrier gas flow-rate. Fast Supersonic GC-MS was able to analyze thermally labile pesticides, such as carbamates, that are difficult or impossible to analyze in standard GC-MS. Large volume injection using a ChromatoProbe was also demonstrated, in the 6 min analysis of pesticides at 20 ng/g in a spice matrix.