Application of GC–MSD and LC–MS/MS for the determination of priority pesticides in baby foods in Serbian market

Babies and small children are especially sensitive population to the exposure to environmental contaminants. Their small mass and developing systems, including brain development may show adverse health effects from even low levels of contamination on a chronic or single dose case. In this paper one extraction method and two chromatographic techniques for the determination of pesticide residues in baby food were evaluated. A liquid chromatography–tandem mass spectrometry technique combined with electrospray ionization (ESI), (LC–MS/MS) and gas chromatography–mass spectrometry detection (GC–MSD) technique were applied in the detection of 50 pesticides in baby food. So-called QuEChERS (quick, easy, cheap, effective, rugged and safe) method was used as a sample preparation procedure. The recoveries were investigated at three levels (5, 10 and 50 μg/kg) and the results obtained showed compliance with the contemporary EU requirements with a few exceptions. LOQs for most of the tested pesticides were below the EU MRLs (10 μg/kg), except deltamethrin, cypermethrin, fenvalerate, phosalone and beta-cyfluthrin (LOQs were 10 μg/kg). Both techniques were applied in the analysis of 50 samples of baby food manufactured in Serbia.     

The application of d-SPE in the QuEChERS method for the determination of PAHs in food of animal origin with GC–MS detection

This paper reports the evaluation of the Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) method for the determination of polycyclic aromatic hydrocarbons (PAHs) in food of animal origin with GC–MS detection. Although in the available literature, there is a lot of information about sample preparation method for PAHs determination in food samples, but the QuEChERS method application for PAHs determination in food of animal origin has not been reported as yet. The results showed that the best recovery ratios 72.4–110.8 % with relative standard deviation lower than 10 % for all determined compounds were received for the method with ethyl acetate as an extraction solvent, primary–secondary amine and C₁₈ sorbents and evaporation to dryness and dissolving the residues in the hexane. The limit of quantification ranged from 0.0003 to 0.0030 mg kg^?1 for pyrene and benzo[a]anthracene, respectively. This method was also used for the determination of PAHs in 15 samples of pork ham. In 8 of 15 samples selected, PAHs were identified. It was observed that in 6 cooked ham and one smoked and cooked samples, any PAHs were found. In other samples, which were smoked and roasted, some low concentration of PAHs was detected. In one sample benzo[a]pyrene (0.0015 mg kg^?1), in one sample benzo[b]fluoranthene (0.0015 mg kg^?1) and in one sample chrysene (0.0024 mg kg^?1) were detected. A number of other less harmful PAHs were also determined. There were no exceedances of maximum levels (according to Commission Regulation (EU) No 835/2011) for determined PAHs in any of the analysed samples.     

Assessment and validation of methods for the determination of γ-glutamyltransferase activity in sheep milk

The determination of enzymatic activity in sheep milk is still today a practically unexplored field of research. This study proposes and fully validates two analytical procedures (i.e. by means of UV–vis spectrophotometry and RP-HPLC methods) for the determination of γ-glutamyltransferase activity in sheep milk. Both methods are characterised by low detection and quantification limits, excellent linearity over a wide enzymatic activity interval, very good repeatability and reproducibility, and are bias-free. The RP-HPLC method provides better sensitivity and automation capability levels than that of UV–vis, and its use is hence suggested for screening purposes. These methods have been preliminarily tested with a number of real samples of whole sheep milk, obtaining an average γ-glutamyltransferase activity value of 2.93 ± 0.50 U ml⁻¹ and a range from 2.72 ± 0.10 U ml⁻¹ to 3.46 ± 0.11 U ml⁻¹.     

Development of an accelerated solvent extraction,ultrasonic derivatisation LC‐MS/MS method for the determination of the marker residues of nitrofurans in freshwater fish

A rapid method using accelerated solvent extraction (ASE) and ultrasound enhanced derivatisation has been developed for the quantitative determination of metabolites of nitrofurans, namely 3‐amino‐2‐oxalidinone (AOZ), 5‐morpholinomethyl‐3‐amino‐2‐oxalidinone (AMOZ), 1‐amino‐hydantoin (AHD) and semicarbazide (SEM), in muscle and skin of carp and finless eel. The target analytes were extracted using ASE, ultrasonic derivatisation for 1?h and then purified by solid phase extraction. Averaged decision limits (CCα) and detection capability (CCβ) of the method were in the range of 0.07–0.13 and 0.31–0.49?µg?kg^?1 in carp and finless eel, respectively. The accuracy in terms of recovery was in the range 77.2–97.4%. The simplified and traditional methods were compared with incurred residue samples. The simplified method reduced the derivatisation time and has been applied to the determination of nitrofurans residues in fish.     

The simultaneous determination of the ionophore antibiotics in animal tissues and eggs by tandem electrospray LC–MS–MS

A method has been developed for the simultaneous extraction and determination of the four most currently used Ionophore antibiotics (lasalocid, monensin, narasin and salinomycin) by LC–MS–MS from different animal tissues and eggs. Results show good repeatability, and mean spiked recoveries for lasalocid, monensin, narasin and salinomycin in animal livers are in the average range 93–103, 96–103, 93–102 and 97–106%, respectively, and in eggs the mean spiked recoveries are 101, 103, 98 and 102% for lasalocid, monensin, narasin and salinomycin, respectively. The detection limit is at 1 ng ml⁻¹ for all the named ionophorous compounds. A quantitation level of 50 ng g⁻¹ for lasalocid, monensin at 2.5 ng g⁻¹, and 10 ng g⁻¹ for narasin and salinomycin is achieved which represents half the action limit prescribed by the UK Regulation in compliance with the European Council Directive 96/23/EC. A high throughput of samples is achievable using this method which allows the analysis of 30–40 samples by one analyst in a day.     

UHPLC–MS/MS highly sensitive determination of aflatoxins,the aflatoxin metabolite M1 and ochratoxin A in baby food and milk

In this work, a method has been developed for the ultrasensitive and selective determination of various regulated mycotoxins (aflatoxins G1, G2, B1, B2, M1, and ochratoxin A) in baby food commodities and milk, using ultra high pressure liquid chromatography (UHPLC) coupled to tandem mass spectrometry (MS/MS). The high sensitivity required for these compounds made necessary the application of a pre-concentration step based on solid phase extraction with immunoaffinity columns, after sample extraction with acetonitrile:water (80:20). Thanks to the fast high-resolution of UHPLC and the enhanced selectivity obtained with the triple quadrupole mass analyser in SRM mode, the chromatographic separation was achieved in only 4 min.     

A method for the determination of acrylamide in a broad variety of processed foods by GC–MS using xanthydrol derivatization

A novel GC–MS method was developed for the determination of acrylamide, which is applicable to a variety of processed foods, including potato snacks, corn snacks, biscuits, instant noodles, coffee, soy sauces and miso (fermented soy bean paste). The method involves the derivatization of acrylamide with xanthydrol instead of a bromine compound. Isotopically labelled acrylamide (d ₃-acrylamide) was used as the internal standard. The aqueous extract from samples was purified using Sep-Pak? C₁₈ and Sep-Pak? AC-2 columns. For amino acid-rich samples, such as miso or soy sauce, an Extrelut? column was used for purification or extraction. After reaction with xanthydrol, the resultant N-xanthyl acrylamide was determined by GC–MS. The method was validated for various food matrices and showed good linearity, precision and trueness. The limit of detection and limit of quantification ranged 0.5–5 and 5–20?µg?kg^?1, respectively. The developed method was applied as an exploratory survey of acrylamide in Japanese foods and the method was shown to be applicable for all samples tested.     

Comparison of headspace-SPME-GC–MS and LC–MS for the detection and quantification of coumarin,vanillin, and ethyl vanillin in vanilla extract products

A headspace-solid phase micro-extraction (HS-SPME) GC–MS method has been developed for the determination of coumarin, vanillin and ethyl vanillin in vanilla products. Limits of detection ranged from 1.33 to 13.2 ng mL⁻¹. Accuracy and precision data for the method were measured and compared to those obtained using LC-ESI-MS. A survey of 24 commercially available vanilla products was completed using both techniques. No coumarin was detected in any of the samples. Examination of the GC–MS chromatograms revealed the presence of 18 other flavor related compounds in the samples. The method validation and sample analysis data using HS-SPME-GC–MS were comparable to those obtained using the LC–MS method. Because the two methods are conceptually different from one another, both methods would not be subject to the same interferences. This would allow them to be used as confirmatory methods for each other.     

A Comparative Study of the Ionization Modes in GC–MS Multi-residue Method for the Determination of Organochlorine Pesticides and Polychlorinated Biphenyls in Crayfish

A gas chromatography–mass spectrometry (GC–MS) multi-residue method with different ionization modes has been developed for the determination of 23 organochlorine pesticides and ten polychlorinated biphenyls (PCBs) in crayfish. The approach was based on an extraction with acetonitrile saturated by hexane (containing 0.1 % acetic acid), followed by a dispersed solid-phase extraction cleanup with primary secondary amine and octadecylsilane, and finally analyzed by GC–MS/MS operated in electron impact (EI) and positive chemical ionization (PCI), and GC–MS in negative chemical ionization (NCI), respectively. An isotope internal standard, DDT-D₈, was used for quantification. These three GC–MS procedures were validated and compared using crayfish samples fortified at three levels (5, 10 and 20 ng/g). Good linearity, accuracy, precision and sensitivity were achieved by all three ionization modes with recoveries in the range of 70 to 112 %, relative standard deviations (RSDs) below 10 % and limits of detection (LODs) in the range of 0.002 to 8 ng/g. Extremely high selectivity was obtained by both EI- and PCI-MS/MS, while PCI was more sensitive to compounds containing nitro and carbonyl groups, and EI was a better choice for dieldrin, endosulfan, and its metabolites. NCI-MS had the highest sensitivity but lower anti-interference ability and was not suitable for the determination of DDTs and PCBs. These three ionization modes can be used as complementary and alternative in routine GC–MS analysis of organochlorine pesticides and polychlorinated biphenyls in food.     

Development of a solid phase microextraction protocol for the GC–MS determination of volatile off-flavour compounds from citral degradation in oil-in-water emulsions

The conditions for headspace solid phase microextraction (HS-SPME) analysis of volatile off-flavour compounds in citral emulsion were determined. Type of SPME phase (65 μm PDMS/DVB, 100 μm PDMS and 75 μm CAR/PDMS), adsorption temperature and salt concentration were significant factors affecting total peak area in the gas chromatogram and optimised in one factor experiments. Then, adsorption temperature (30–50 °C), adsorption time (20–40 min), and salt concentration (0–6 M) were studied to develop HS-SPME condition for obtaining the highest extraction efficiency. PDMS/DVB in 65 μm was the optimum fiber because of high adsorption efficiency and good reproducibility. The optimal condition was adsorption at 50 °C for 40 min and 6 M salt added to sample. Good Linearity, high recovery, good reproducibility and low limit of detection (LOD) for all off-odour compounds according to the optimised SPME conditions indicated that the SPME procedure was applicable for the analysis of the degraded citral products in headspace volatile of emulsion.     

Effervescent-assisted dispersive liquid–liquid microextraction based on the solidification of floating organic droplets for the determination of fungicides in vinegar and juice

ABSTRACT

A method of effervescent-assisted dispersive liquid–liquid microextraction based on the solidification of a floating organic droplet is reported. This approach was used to measure the fungicides myclobutanil, tebuconazole and epoxiconazole in vinegar and juice. Acidic vinegar and juice reacted with the carbonate to produce effervescence in situ, which then promoted contact of the sample with the extraction solvent. The 1-dodecanol extraction solvent helped solidify the floating organic droplets and could be fully dispersed by effervescence. The extraction solvent, salt, carbonate, and extraction time were optimised. The optimal conditions were 200 μL of 1-dodecanol, 100 mg of sodium chloride, 50 mg of sodium bicarbonate, and 30 s of extraction time. The proposed method has good linearity between 10 and 1000 ng mL^?1. Recoveries were between 70.4% and 103.1% in different vinegar and juice samples. This method was successfully used to measure fungicides in vinegar and juice. This system is simple, fast and environmentally friendly.     

Development and validation of a new LC–MS/MS method for the simultaneous determination of six major ergot alkaloids and their corresponding epimers. Application to some food and feed commodities

A liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous determination of six major ergot alkaloids, ergometrine, ergosine, ergotamine, ergocornine, ergokryptine and ergocristine, as well as their corresponding epimers in food and feed samples. The method involves extraction under alkaline conditions and subsequent clean-up by applying a simple and rapid liquid–liquid partitioning procedure prior to LC–MS/MS analysis. Evaluation of the method revealed good linearity, accuracy and precision. The limits of quantification varied from 0.1 to 1 μg/kg depending on the analyte and matrix. The average extraction and clean-up recoveries in different matrices were between 45 (only for ergometrine in biscuit) and 90%. The uncertainty associated with the analytical method was not higher than 51% and 30%, at concentration levels of 2.5 and 150 μg/kg respectively. Analyte epimerization proved to be minimal during the analytical procedure. The method has been successfully applied to the determination of ergot alkaloids in some Belgian food and feed commodities. Ergot alkaloids were found in 104 out of 122 samples investigated. Ergosine was the most frequently occurring alkaloid, while the highest levels were observed for ergotamine, ergocristine or ergosine, depending on the product type. The total alkaloid content in positive samples varied from 1 to 1145 μg/kg.     

Liquid chromatography–tandem mass spectrometry for the determination of chrysoidine in yellow-fin tuna

A high performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry (MS/MS) method was described for the residue detection of chrysoidine in yellow-fin tuna in the present study. Samples were cleaned up with solid phase extraction (SPE) cartridge, and then injected into HPLC for separation. Multiple-reaction monitoring (MRM) was applied for quantitative determination. Results showed that the low limit of detection (LOD) of the method was 1.25 × 10⁻¹² g, and the low limit of quantification (LOQ) was 0.42 μg/L. The standard calibration curve was y = 2333.9x −845 (r² > 0.99) with the linear range of 0.63–100 μg/L. The average recoveries of chrysoidine ranged from 86.0% to 108.0% when the spiked concentration was from 0.5 μg/kg to 20 μg/kg. And the developed method also showed the good test precisions (RSD%: 4.38–14.27%).     

The UPLC–ESI–QqQLIT–MS/MS method for quantitative determination of phytochemicals in ethanolic extracts of different parts of eight Ficus species: Development and validation

Ficus and validation of the ultra performance liquid chromatography–electrospray ionization hybrid triple quadrupole–linear ion trap–tandem mass spectrometry (UPLC–ESI–QqQ_LIT–MS/MS) method in a multiple-reaction monitoring (MRM) mode for the quantitative determination of 19 phytochemicals. The chromatographic separation of targeted phytochemicals was performed using the Waters ACQUITY UPLC BEH? C18 column (1.7 μm, 2.1 mm × 50 mm) with 0.1% formic acid with water and acetonitrile as a mobile phase at a flow rate of 0.25 mL/min. The validation parameters showed the overall recoveries from 95.78?101.44% (RSD ≤ 3.25%), precision (intra-day: RSD ≤ 2.96%; inter-day: RSD ≤ 2.89%), linearity (R² ≥ 0.9982), limit of detection (8.60 × 10^–10?2.18 × 10^–6 mg/mL), and the limit of quantitation (2.60 × 10^–9–6.63 × 10^–6 mg/mL) in the concentration range from 0.5 to 1000 × 10^–6 mg/mL. This method was successfully applied in ethanolic extracts of different parts (fruits, leaves, and barks) of selected eight Ficus species. Quinic acid was predominant followed by rutin and chlorogenic acid among the studied nineteen phytochemicals. Ficus benjamina showed the maximum total content in fruits and leaves. The UPLC–ESI–QqQ_LIT–MS/MS method combined with principal component analysis (PCA) was successfully used for Ficus species discrimination on the basis of the contents of 15 compounds. The UPLC–ESI–QqQ_LIT–MS/MS method combined with PCA could be used for quality control.     

Monitoring of the amphetamine-like substances in dietary supplements by LC-PDA and LC–MS/MS

Recently, amphetamine-like substances derived from the β-phenylethylamine core structure have been detected in dietary supplements. Especially, β-methylphenylethylamine (BMPEA), an amphetamine isomer, has been found in dietary supplements labeled as containing Acacia rigidula. The U. S. Food and Drug Administration determined that BMPEA is not naturally present in food and does not meet the statutory definition of a dietary ingredient. In addition, BMPEA has been classified as a psychotropic drug in South Korea and a doping substance by the World Anti-Doping Agency. The aim of this study was to determine whether dietary supplements contained amphetamine and amphetamine-like substance, including β-phenylethylamine (β-PEA) and BMPEA using LC-PDA and LC–MS/MS. In 10 of 110 samples, illegally added compounds were detected in the following ranges; β-PEA 1.4–122.0 mg/g and BMPEA 4.7–37.6 mg/g. This study will contribute to enhancement of food safety in the South Korea.     

Acetonitrile: the better extractant for the determination of T-2 and HT-2 toxin in cereals using an immunoaffinity-based cleanup?

The complete extraction of the analytes is the most important item during the analysis of matrix samples. Within the discussion, which of both extractants—aqueous methanol or aqueous acetonitrile—is the best for the extraction of the mycotoxins T-2 and HT-2, a recently published work pointed out, that the latter one resulted in incorrect high toxin concentrations. Due to water absorption of dry samples and salting out effects, a phase separation of the extractant occurs and the toxins accumulate in the organic layer. Based on these results, investigations were performed by comparing and evaluating the extraction efficiency of methanol/water and acetonitrile/water—the most frequently used extractants for T-2 and HT-2 toxin. Therefore, a reliable and routine capable method combining the extraction with acetonitrile/water and a cleanup with immunoaffinity columns was necessary. Due to the sensitivity of the antibodies against acetonitrile, the cleanup of extracts containing acetonitrile was critical. A modified workup based on the investigated antibodies’ tolerance of 5% of acetonitrile was established. The method was validated according to common guidelines (linearity, limit of detection/limit of quantitation, accuracy, method precision, recovery). Especially for more complex matrices a better extraction efficiency with acetonitrile/water was obtainable, simple or unprocessed matrices resulted in identical toxin concentrations independent from extraction solvent.     

Nicotine determination in mushrooms by LC–MS/MS with preliminary studies on the impact of drying on nicotine formation

A problem concerning significant amounts of nicotine in dried wild mushrooms (mainly Boletus edulis from China) has been reported to the European Commission. As a consequence, the European Food Safety Authority (EFSA) proposed temporary maximum residue levels (MRLs) of 0.036 mg kg^?1 for fresh wild mushrooms and 1.17 mg kg^?1 for dried wild mushrooms (2.3 mg kg^?1 for dried ceps only). The EFSA also highlighted the necessity for a monitoring and testing programme to be launched by food business operators at the start of the 2009 harvest season. In the present study, a quick and sensitive analytical method for routine analysis of nicotine in fresh and dried mushrooms was developed and validated by a single-laboratory procedure. The method, which employs an LC–MS/MS system and (±)-nicotine-d ₄ as internal standard, has a limit of quantification of 6 and 60 µg kg^?1 for fresh and dried product, respectively. Analyses of samples spiked with different levels of nicotine showed recoveries ranging from 107 to 122%, with relative standard deviations of 2.9–10.1% depending on the spiking level. The combined uncertainties, calculated at a low level for frozen (0.015 mg kg^?1) and a high level for the dried (2 mg kg^?1) matrix, were 13 and 10%, respectively. Application of the method to real samples of mushrooms purchased on the market or obtained from local producers showed nicotine levels ranging 0.01–0.04 and 0.1–4.5 mg kg^?1 in fresh/frozen and dried matrices, respectively. To establish reasons for the unexpectedly high levels of the nicotine in dried matrices, preliminary laboratory experiments involving drying mushrooms were performed under various conditions.     

Tandem solid phase extraction coupled to LC–ESI–MS/MS for the accurate simultaneous determination of five heterocyclic aromatic amines in processed meat products

Carcinogenic heterocyclic aromatic amines are difficult to measure since only trace levels are present in processed meat products. In this study, typical heterocyclic aromatic amines, including 2-amino-3-methylimidazo [4,5-f] quinoline (IQ), 2-amino-3,4-dimethylimidazo [4,5-f] quinoline (MeIQ), 2-amino-3,8-dimethyli-midazo [4,5-f] quinoxaline (MeIQx), 2-amino-3,4,8-trimethylimi-dazo [4,5-f] quinoxaline (4,8-DiMeIQx), 2-amino-1-methyl-6-ph-enylimidazo [4,5-b]pyridine (PhIP), were studied to develop a sensitive and accurate method for their rapid quantification in animal-derived products, with 2-Amino-3,4,7,8-dimethylimidazo [4,5-f] quinoxalline (TriMeIQx) as an internal standard. Liquid chromatography–electrospray-tandem mass spectrometry conditions were analyzed to enhance detection sensitivity. Diatomaceous earth was employed to extract heterocyclic aromatic amines from meat samples, and the analytes were purified and enriched using tandem solid phase extraction, with siliprep propylsulfonic acid coupled to a C₁₈ cartridge. A number of parameters, including pH, eluent and volume, were carefully optimized to improve the extraction and purification efficiency. Under the optimal experimental conditions, the limits of detection for each analyte within the meat matrix were 0.5 pg (injected). The established method was applied to evaluate commercial meat products. At three spiked levels of 0.2, 1 and 4 μg kg⁻¹, the recoveries and relative standard deviations were measured as 76.4–122.2 and 0.9–23.4%, respectively, suggesting the developed method is promising for the accurate quantification of heterocyclic aromatic amines at trace levels in processed meats.     

A versatile method for the quantitative determination of β N-alkanoyl-5-hydroxytryptamides in roasted coffee

Since literature reports suggest N-alkanoyl-5-hydroxytryptamides to account for stomach irritations perceived by sensitive subjects after coffee consumption, there is increasing interest in the concentrations of these substances in coffee and coffee products. A versatile analytical method for the accurate quantitative analysis of N-alkanoyl-5-hydroxytryptamides in coffee by means of reversed-phase high-performance liquid chromatography with fluorescence detection and synthetic N-heptadecanoyl-5-hydroxytryptamide as the internal standard was developed. Spiking coffee samples with known amounts of N-alkanoyl-5-hydroxytryptamides followed by quantitative analysis revealed recovery rates of 99–103%. On the basis of the results of liquid chromatography–mass spectometry/mass spectometry experiments as well as co-chromatography with the synthetic reference compounds, the previously unknown N-heneicosanoyl-5-hydroxytryptamide and N-tricosanoyl-5-hydroxytryptamide were detected for the first time in coffee.