A reliable screening of mycotoxins and pesticide residues in paprika using ultra-high performance liquid chromatography coupled to high resolution Orbitrap mass spectrometry

In this study, an ultra-high performance liquid chromatography (UHPLC) coupled to a high resolution Orbitrap mass spectrometry (Orbitrap-HRMS) was demonstrated as a promising technique in high-throughput method development for the routine analysis and contamination control of mycotoxins and pesticide residues in spices. The method was applied for the analysis of fifty ground paprika samples containing blends of sweet and hot paprika harvested in Brazil and China. The efficiency and detection sensitivity of the used UHPLC-Orbitrap-HRMS technique were compared to the results obtained using a triple quadrupole tandem mass spectrometric detector (UHPLC-QqQ-MS/MS). The values of recovery (75–120%) and repeatability (8–15%) for both methods, calculated as the average (n = 5) from the results of spiked (10–500 μg kg⁻¹), paprika samples, were in good conformity to the relevant EU guidelines. The high resolution of the used Orbitrap-HRMS technique provided a better sensitivity for quantitative determination of several pesticide contaminants in paprika, compared to the results obtained by the QqQ-MS/MS method and were comparable in case of mycotoxins. The results of analysis demonstrated the ubiquitous presence of three mycotoxins (fumonisin B₁, ochratoxin A, and sterigmatocystin) and twelve pesticide residues in paprika. The concentrations of determined contaminants were below the MRLs set by the Regulations of the European Union with exception of iprovalicarb, which violated the EU MRL in two samples of hot paprika. In addition, a notable difference in the concentration of fumonisin B₁ was determined depending on the harvest period (2009–2013), reaching the maximum concentrations of 33 μg kg⁻¹ in sweet paprika and 140 μg kg⁻¹ in hot paprika. There was no significant correlation found between the determined mycotoxin contamination levels and the pesticide residues, with the sole exception of decreased fumonisin B₁ content in samples with an elevated concentration of metalaxyl fungicide.     

Rapid screening and quantification of multi-class multi-residue veterinary drugs in royal jelly by ultra performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry

A simple and rapid multi-class multi-residue analytical method was developed for the screening and quantification of veterinary drugs in royal jelly by ultra performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). A total of 90 veterinary drugs investigated belonged to more than 14 families such as lincomycins, macrolides, sulfonamides, quinolones, tetracyclines, β-agonists, β-lactams, sedatives, β-receptor antagonists, sex hormones, glucocorticoids, nitroimidazoles, benzimidazoles, nitrofurans, and the others. A modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) procedure was used for the sample preparation without solid-phase extraction step. The linearity, sensitivity, accuracy, repeatability, and reproducibility of the method were fully validated. The response of the detector was linear for each target compounds in wide concentration range (at least, two orders of magnitude) with correlation coefficient (R²) of 0.9921–0.9999. The range of the limit of quantification for these compounds in the royal jelly ranged from 0.21 to 20 μg/kg. The repeatability and reproducibility were in the range of 3.01–11.6% and 5.97–14.9%, respectively. The average recoveries ranged from 70.21 to 120.1% with relative standard deviation of 1.77–9.90% at three concentration levels. For the screening method, the data of the precursor and product ions of the target analytes were simultaneously acquired under the All Ions MS/MS mode in a single run. A homemade database including the elemental composition, accurate masses, retention time, isotopic pattern data of the target ions the characteristic in-source fragment ions was utilized for the confirmation and identification of the target compounds. The applicability of the screening method was verified by applying to real royal jelly samples, and certain veterinary drugs were detected in some cases.     

Method for analysis of 68 organic contaminants in food contact paper using gas and liquid chromatography coupled with tandem mass spectrometry

Food contact materials (FCMs) have been reported as a source of various xenobiotics. This study investigates the possibility of simultaneous analysis of 68 potential contaminants in paper FCMs, specifically phthalates, polycyclic aromatic hydrocarbons, photoinitiators, bisphenols, and polyfluorinated compounds. Target compounds were co-isolated using a technique based on ultrasonic extraction by mixture of acetonitrile and water followed by QuEChERS-like liquid–liquid partition in the presence of inorganic salts. Resulting extracts were analyzed using gas and high performance liquid chromatography coupled with tandem mass spectrometry (GC–MS/MS, HPLC–MS/MS). Acceptable recoveries (70–120%) and RSDs (<20%) were achieved for most of the analytes at spiking levels of 0.05, 0.2 and 1 mg/kg. LOQs ranged from 0.0013 to 0.22 mg/kg. The proposed method was successfully applied to analysis of 15 real samples. Complex mixtures of contaminants reaching levels up to 48 mg/kg were identified in the samples.     

Simultaneous determination of Fusarium mycotoxins in wheat grain from Morocco by liquid chromatography coupled to triple quadrupole mass spectrometry

In the present study, eighteen (18) mycotoxin produced by the genus Fusarium (fumonisins (FB1, FB2 and FB3), type-B trichothecenes (NIV, DON, FUS-X, 3Ac-DON and 15Ac-DON), type-A trichothecenes (NEO, DAS, T2 and HT2), zearalenone (ZEA), beauvericin (BEA), and enniatins (ENA, ENA₁, ENB and ENB₁)) were monitored in different samples of wheat grain commercialized in Morocco. A liquid chromatography coupled to triple quadrupole mass spectrometry method previous matrix solid phase dispersion extraction was used for sample analysis. A total of eighty (80) samples of durum wheat were collected in different local markets from several areas in Morocco. Analytical results showed that 54 out of 80 total wheat samples (68%) were contaminated. The mycotoxins found were ENA, ENA₁, ENB and ENB₁, DON and BEA. The rest of investigated mycotoxins were below the limits of quantification. In positive samples, enniatins levels ranged between 2.5 and 2570 μg/kg. DON levels ranged between 121 and 1480 μg/kg and BEA levels were between 5.4 and 16 μg/kg. Among enniatins, ENB was predominant with a frequency of 61%. Co-presence of DON, enniatins and BEA from durum wheat from Morocco was found for the first time.     

Rapid screening and quantification of cyromazine,melamine, ammelide,ammeline, cyanuric acid,and dicyandiamide in infant formula by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and triple quadrupole mass spectrometry

In the past few years, special concern for triazine pesticide cyromazine (CYRO) and its metabolites melamine (MM), ammelide (AMD), ammeline (AMN), cyanuric acid (CA), and dicyandiamide (DCD) residues has arisen in the food safety field on infant foods. In this work, screening and confirmation for infant milk powder samples were performed by quadrupole time-of-flight mass spectrometry (QTOF-MS). An accurate mass database was established. The matrix-matched internal standard calibration curves of UPLC–MS/MS were linear, with correlation coefficient (r²) higher than 0.994. The limits of quantification (LOQ) were 1.6, 5.0, 16.5, 10.0, 66.4, and 66.4 μg/kg for CYRO, MM, AMD, AMN, CA, and DCD, respectively. The average recoveries ranged from 64.8 to 103%. The retention time, intra- and inter-day precisions (relative standard deviation, RSDs) of 6 analytes were in the range of 0.02–0.05%, 2.4–6.1%, and 7.2–11.2%, respectively. CYRO and MM residues in some infant milk powder samples were confirmed. Quantitation of positive samples was performed using the internal standard method and six-point matrix-matched calibration curves. The content for CYRO in all 8 samples and MM in 4 samples were in the range of 3.5–45 μg/kg and 8–25 μg/kg, respectively. The method developed for the first time has high sensitivity, can be used as an effective tool for rapid screening and highly sensitive multiresidue determination of CYRO, MM and their metabolites AMD, AMN, and CA as well as DCD in milk powder samples.     

Method validation for 243 pesticides and environmental contaminants in meats and poultry by tandem mass spectrometry coupled to low-pressure gas chromatography and ultrahigh-performance liquid chromatography

An easy and reliable high-throughput analysis method was developed and validated for 192 diverse pesticides and 51 environmental contaminants (13 PCB congeners, 14 PAHs, 7 PBDE congeners, and 17 novel flame retardants) in cattle, swine, and poultry muscle. Sample preparation was based on the “quick, easy, cheap, effective, rugged and safe” (QuEChERS) approach using filter-vial dispersive solid-phase extraction (d-SPE) cleanup. Split final extracts were analyzed in parallel by low-pressure (vacuum outlet) GC-MS/MS and UHPLC-MS/MS (10 min each), providing an additional degree of confirmation for 55 overlapping LC- and GC-amenable pesticides. Analyte protectants were utilized to improve sensitivity and decrease matrix effects in GC analysis, and only filtration of initial extracts was enough to avoid ion suppression in UHPLC-MS/MS. The method was validated at three spiking levels (10, 25, and 100 ng/g) at or below established tolerance levels in the sample types. Satisfactory recoveries (70–120%) and RSDs ≤ 20% were achieved for 200 analytes. The validated method was successfully applied to the analysis of real-world incurred meat samples, further demonstrating the utility of the method for implementation in regulatory and commercial laboratories.     

A liquid chromatography mass spectrometry-based method to measure organophosphorous insecticide, herbicide and non-organophosphorous pesticide in grape and apple samples

The LC-MS/MS with Quick, Easy, Cheap, Effective, Rugged and safe method was used for analysis of eighteen pesticides in fruit samples. This method was found to be accurate (≥99%), as it possessed limits of detection in the 0.002-0.087 ranges respectively. The coefficients of variations (≥0.9999) were less than 2% at the low ng g⁻¹ concentration. Mean recoveries ranged between 97 and 101%, and % RSD were below 5%. The imidacloprid mean concentrations of red grapes (125.124 ng g⁻¹) and green grapes (702.030 ng g⁻¹) differed significantly (p < 0.05) between the grapes. Similarly, the fenitrothion mean concentration of red grapes (143.66 ng g⁻¹) and green grapes (51.554 ng g⁻¹) differed significantly (p < 0.001) between the fruits. The average concentration of quinalphos was 4.317 and 1.389 ng g⁻¹ differed significantly (p < 0.01) between the grapes. In apples imidacloprid, quinalphos, triazophos, ethion and acephate were also present. This study may be helpful in developing a regional exposure database and in the facilitation of health risk assessment due to pesticide exposure.     

Determination of chloramphenicol,thiamphenicol, and florfenicol in fish muscle by matrix solid-phase dispersion extraction (MSPD) and ultra-high pressure liquid chromatography tandem mass spectrometry

An UPLC–MS/MS method based on matrix solid-phase dispersion (MSPD) was developed for the determination of chloramphenicol (CAP), thiamphenicol (TAP) and florfenicol (FF) in fish muscle. Muscle tissue was blended with octadecylsilyl-derivatized silica (C₁₈). A column made from the C₁₈/muscle tissue matrix was washed with n-hexane, after which CAP was eluted with acetonitrile/water (50:50, v/v) and partitioned into ethyl acetate. The final extract was evaporated, and reconstituted into methanol/water (10:90, v/v) for the analysis of UPLC–MS/MS. The correlation coefficient (r²) with each matrix-matched calibration curve is higher than 0.999. CCα and CCβ of the fenicols upon the method were ranged from 0.02 to 0.06 μg kg⁻¹ and 0.11–0.16 μg kg⁻¹ respectively. At the fortified levels, mean recoveries of the three compounds were ranged from 84.2% to 99.8%, and the RSDs were lower than 12%. The method was accurate and reproducible, being successfully applied to the monitoring of CAP, TAP and FF in fish samples obtained from the Chinese market.     

High performance liquid chromatography method for the determination of patulin and 5-hydroxymethylfurfural in fruit juices marketed in Malaysia

A rapid, simple, improved method for the simultaneous determination of patulin (PAT) and 5-hydroxymethylfurfural (5-HMF) in fruit juices is described. The target compounds were extracted with ethyl acetate using vortex followed by high performance liquid chromatographic separation. PAT and 5-HMF were separated within 4 min using Poroshell C18 column with acetonitrile:water (1:9, v/v) as the mobile phase. Under the optimized conditions, the detection limits of PAT and 5-HMF were 0.25 and 0.46 ng mL⁻¹, respectively while the quantification limits were 0.76 and 1.40 ng mL⁻¹, respectively. The recoveries of PAT and 5-HMF at 50, 750 and 5000 ng g⁻¹ ranged from 92.8 to 108%. The proposed method was applied to fivety-six fruit juices (apple, mango, pineapple, guava, lychee, tamarind, soursop and mixed fruit) and PAT was found in three samples ranging from 13.1–33.7 ug L⁻¹. 5-HMF was found in all the samples ranging from 0.08 to 91.5 mg L⁻¹. Liquid chromatography-tandem mass spectrometry with triple quadrupole analyzer was used to confirm the presence of 5-HMF and PAT in some of the contaminated samples.     

Dual-dummy-template molecularly imprinted polymer combining ultra performance liquid chromatography for determination of fluoroquinolones and sulfonamides in pork and chicken muscle

In this study, a dual-dummy-template molecularly imprinted polymer capable of simultaneous recognizing 8 fluoroquinolones and 8 sulfonamides was synthesized. Its recognition performance was investigated by comparing the 3D conformations of four dummy templates and the two classes of drugs based on computational simulation. Then a solid phase extraction column was prepared and optimized that was combined with ultra performance liquid chromatography for determination of the 16 drugs in pork and chicken. The column could be reused for at least eighty times, and it showed high absorbency capacities (34.9–74.2 μg) and high recoveries (92%–99%) to these drugs. The limits of detection of this method for the two classes of drugs in meat were in the range of 1.0–3.4 ng/g, and the recoveries from the standards fortified blank samples were in the range of 86.1%–109.4%. Therefore, this method could be used as a specific, sensitive and accurate method for determination of fluoroquinolones and sulfonamides in meat.     

原油中的石油酸组分全二维气相色谱/飞行时间质谱分析

通过分析原油酸性组分衍生化样品,初次尝试了利用全二维气相色谱/飞行时间质谱技术（GC×GC-TOFMS）鉴定原油有机酸性化合物组成。研究结果表明：检测的原油中含有脂肪酸和环烷酸2类酸性化合物,其中脂肪酸类化合物主要由正构一元脂肪酸、类异戊二烯酸、2-乙基链烷酸和正构二元脂肪酸构成;环烷酸类化合物主要由单环长链脂肪酸、1,4-二甲氧基蒽、脱氢松香酸和甾烷酸等化合物构成。全二维气相色谱飞行时间质谱为石油酸组分的精细地球化学剖析提供了新的分析手段。     

Investigation of different sample pre-treatment routes for liquid chromatography–tandem mass spectrometry detection of caseins and ovalbumin in fortified red wine

Different sample treatment protocols for the liquid chromatography–electrospray-tandem mass spectrometry (LC–ESI-MS/MS) analysis of potential residuals of ovalbumin and caseins added to red wines were developed. In particular, attention was paid to the simultaneous detection and quantitation of fining agent residues, i.e. ovalbumin, α- and β-casein, in wine samples. The different sample treatment methods were compared in terms of protein recovery. The use of denaturing agents combined with size exclusion concentration and purification allowed to obtain a reproducible (RDS < 20%) analytical protocol with good recoveries (73(±2) – 109(±4)% range) for digested proteins from 12.5 mL of wine sample. Matrix-matched calibration from LC–ESI-MS/MS analysis indicated that the devised method allowed detection of target peptides in the 0.01–0.8 μg/mL range. Finally, method applicability and selectivity was demonstrated by using fining agents commonly exploited in winery industry and by analyzing 20 commercial red wine samples.     

高效液相色谱法分析车用料挥发性醛酮类化合物

使用2,4-二硝基苯肼(DNPH)采样管,建立了一种高效液相色谱(HPLC)法分析车用料挥发性醛酮类化合物的方法。结果表明:经优化得到分离度最佳的HPLC分析条件为:选用Analytical C 18型色谱柱为分离柱,以乙腈和水为流动相,V(乙腈)/V(水)为70/30,柱温为45℃,流速为1.0 mL/min,进样量为25μL;一次进样能够使13种挥发性醛酮类物质有效分离,完成1个试样的检测时长约为25 min;对于中国石油兰州石化公司的车用料SP 179,在所得最佳HPLC分析条件下,13种醛酮类化合物的质量浓度在0.045~3.000 mg/L内各组分标准曲线的相关系数均等于1,具有良好的线性关系,且各组分的分离度均大于1.1;而且,当在DNPH管添加3个不同质量浓度为0.03,0.06,0.09 mg/L的13种醛酮的混合标准溶液标样后,车用料SP 179中的DNPH采样加标回收率为96.67%~104.44%,3次重复加标回收测定结果的相对标准偏差为0.92%~6.90%。该方法快捷、准确可靠,可适用于车用料中挥发性醛酮类化合物的检测分析。     

Targeted approach for qualitative screening of pesticides in salmon feed by liquid chromatography coupled to traveling-wave ion mobility/quadrupole time-of-flight mass spectrometry

The current work reports the development and validation of a qualitative screening method for 156 pesticides in Atlantic salmon feeds based on the use of ultrahigh-performance liquid chromatography (UHPLC) coupled to traveling-wave ion mobility spectrometry (TWIMS)/quadrupole time-of-flight mass spectrometry (QTOFMS). This novel hyphenation allows a straightforward estimation of the collision cross section (CCS), a physicochemical property that recently has been proposed as an additional identification point for pesticides.In order to increase the overall efficiency of the screening workflow, data processing parameters and tolerances involved in the automated analyte detection were first optimized. The use of the CCS as an additional identification point greatly improved the method selectivity, decreasing the rate of false-positive detections in blank feed samples. Qualitative validation was then performed in accordance with the EU document SANTE/11945/2015 on analytical quality control and validation procedures for pesticide residues analysis in food and feed. Feed samples were spiked with 156 pesticides at three levels (0.01, 0.05 and 0.20 mg/kg) and analyzed by the proposed method. Under the optimized conditions, screening detection limits (SDLs) ≤0.05 mg/kg were achieved for 82% of the investigated pesticides.The applicability of this new approach for the screening of pesticides was finally proven by analyzing several commercial Atlantic salmon feed samples as well as various plant-based feed ingredients. Pirimiphos-methyl and ethoxyquin were among the most frequently detected analytes in the investigated samples.     

Rapid multiresidue and multi-class screening for antibiotics and benzimidazoles in feed by ultra high performance liquid chromatography coupled to tandem mass spectrometry

An analytical strategy was developed for high-throughput screening of multiple antibiotics and two benzimidazoles in feed. Generic sample processing was applied without any purification step. After methanol extraction, the samples were centrifuged, concentrated, and analysed by ultra-high-performance liquid chromatography hyphenated to tandem mass spectrometry in the multiple reaction monitoring mode. Qualitative validation was carried out for more than 50 antibacterials of various classes, including aminocoumarin, amphenicols, beta-lactams, lincosamide, macrolides, diaminopyrimidine, quinolones, sulfonamides, streptogramin, pleuromutilin, polypeptide, quinoxaline, and tetracyclines, and also some benzimidazoles in feed at μg/kg level. Validation was done in accordance with the guidelines laid down in European Commission Decision 2002/657/CE for qualitative screening methods.This convenient, reliable, and sensitive method has been used successfully to monitor antibiotic residues in feeds.     

Simultaneous determination of ethyl carbamate,chloropropanols and acrylamide in fermented products,flavoring and related foods by gas chromatography–triple quadrupole mass spectrometry

A simultaneous determination method was established by gas chromatography–triple quadrupole mass spectrometry (GC–MS/MS) for ethyl carbamate, acrylamide and chloropropanols (1,3-dichloropropane-2-ol (1,3-DCP), 2,3-dichloropropane-1-ol (2,3-DCP), 3-monochloropropane-1,2-diol (3-MCPD) and 2-monochloropropane-1,3-diol (2-MCPD)) in fermented products, flavoring and related foods. Ethyl carbamate-d₅, 1,3-DCP-d₅, 3-MCPD-d₅ and acrylamide-d₃ were used as isotope internal standards for ethyl carbamate, DCPs, MCPDs and acrylamide, respectively. The sample was extracted and cleaned in one single step by a combined clean-up column packed with Extrelut™ NT and primary secondary amine (PSA). The concentrated extract was directly injected without derivatization, separated by an Innowax (cross-bonded polyethylene glycol) capillary column and measured by GC–MS/MS. The calculated limits of detection (LODs) in the sample matrix were 1.8, 1.0, 2.1, 5.3, 5.1 and 5.0 μg kg⁻¹ for ethyl carbamate, 1,3-DCP, 2,3-DCP, acrylamide, 3-MCPD and 2-MCPD, respectively. The average recoveries in different matrices at three spiked concentration levels (20, 100 and 400 μg kg⁻¹) were 88.6–112, 92.6–103, 92.1–106, 86.2–107, 90.4–109 and 91.0–105% with relative standard deviations (RSDs) (Average RSDs in bracket) of 4.4–10.2 (6.4%), 3.7–7.3 (5.4%), 3.9–7.7 (5.2%), 4.4–9.3 (6.4%), 4.1–10.7 (6.3%) and 3.7–7.9% (5.6%), respectively. Two to six contaminants were simultaneously found in some of the soy sauce samples. Ethyl carbamate, acrylamide, 3-MCPD and 2-MCPD were found in the instant noodle flavoring. Trace levels (<12 μg kg⁻¹) of ethyl carbamate, acrylamide or 3-MCPD were detected in tomato sauce and salad dressing. Both ethyl carbamate and acrylamide were found in some of the curry products and Chinese yellow rice wine. Only ethyl carbamate was found in beer and grape wine.     

Simultaneous analysis of 10 polycyclic aromatic hydrocarbons in roasted coffee by isotope dilution gas chromatography-mass spectrometry: Optimization,in-house method validation and application to an exploratory study

The presence of polycyclic aromatic hydrocarbons (PAHs) in food implies a potential risk to human health. Although the coffee has been appointed as an important matrix to be monitored for contamination by PAHs, there is still a gap in the literature regarding to the methods of analysis appropriately validated for this scope and also related to the occurrence of contamination in Brazilian commercial samples. In this paper, a method for the determination of ten priority PAHs in roasted coffee by isotope dilution gas chromatography coupled to mass spectrometry was optimized, validated and applied to an exploratory study with 24 commercial samples. The samples were extracted with simultaneous clean-up using pressurized liquid extraction (PLE), followed by liquid–liquid extraction (LLE) and clean-up using silica columns. Linearity was obtained in the concentration range from 0.25 to 4.00 μg kg⁻¹. Matrix effects were not significant. The precision was demonstrated with relative standard deviations varying from 3.3 to 24 % under repeatability and from 3.3 to 33 % under intermediate precision conditions. The average recovery values ranged between 87 and 111%. These values, as well as the limits of detection, limits of quantification and uncertainties of measurements, met the performance criteria established by European Union regulations. The contamination of the commercial samples ranged from 1.00 ± 0.35 to 11.29 ± 2.33 μg kg⁻¹, indicating the need of further occurrence studies and establishment of monitoring programs in Brazil.