Orbital Ion Trap Mass Spectrometry Research Articles

Exposure Level and Risk Impact Assessment of Pesticides and Veterinary Drugs in Aquaculture Environment

To explore the exposure level of pesticides and veterinary drugs in an aquaculture environment and its impact on the ecological environment, this study took the aquaculture environment in Shanghai as an example, and samples of water, sediment, and inputs from 40 major aquaculture farms were collected from July to September 2022. The types and contents of pesticides and veterinary drugs were screened using high-performance liquid chromatography-electrostatic field orbital ion trap mass spectrometry, and the risk quotient (RQ) method was used to assess the ecological risk of pesticide contamination in water and sediment. The results showed that 13 drugs were screened out from 204 samples (72 samples of water, 72 samples of mud, and 60 samples of input), namely, chlorpromazine, carbendazim, thiophanate, diazepam, florfenicol, simazine, amantidine, diazepam, trimethoprim, ciprofloxacin, ofloxacin, mebendazole, and enrofloxacin. Among them, 12 species were found in water samples with concentrations ranging from 0.016 μg·L-1 to 2.084 μg·L-1. The concentrations of seven species in the mud samples ranged from 0.018 μg·kg-1 to 23.101 μg·kg-1. The results showed that there were four types of inputs, ranging from 1.979 μg·kg-1 to 101.940 μg·kg-1. Seven drugs were found in both water and sediment. The risk quotient (RQ) results showed that there were some high and middle risks in both water and sediment samples of aquaculture farms, and the ecological risks of carbendazim were the highest in both water and sediment samples of aquaculture farms; the RQ values were 3.848 and 1.580, respectively, indicating high risk. It is suggested to strengthen the control and management of exogenous pesticides and veterinary drugs in aquaculture environments to protect the ecosystem health of the aquaculture environment.

Determination of steroid hormone residues in farmed fish using high-resolution orbital ion trap mass spectrometry.

A method for the determination of 15 steroid hormones in farmed fish by liquid chromatography-orbital ion trap mass spectrometry has been developed and validated. The method involved sample preparation with acetonitrile extraction and clean-up, and separation in the LC using a C18 column. The orbital ion trap MS was operated at a resolution of 35 000 FWHM in selected ion monitoring mode. An ion source with heated electrospray ionization was used in positive ionization mode. The samples were prepared by solid-phase extraction. The limit of quantification of steroid hormones in fish samples was 2 μg kg-1. Good linearity was observed since correlation coefficients were more than 0.99 for all compounds. Recoveries of spiked fish samples (2 μg kg-1 and 20 μg kg-1) ranged from 80.8% to 112.6% with relative deviations less than 15%. The method was successfully applied to detect steroid hormones in real farmed fish samples at the μg kg-1 level.

Front Cover: Automated micro‐solid‐phase extraction clean‐up of polycyclic aromatic hydrocarbons in food oils for analysis by gas chromatography–orbital ion trap mass spectrometry

Front Cover: Automated micro‐solid‐phase extraction clean‐up of polycyclic aromatic hydrocarbons in food oils for analysis by gas chromatography–orbital ion trap mass spectrometry

N-nitrosoethylenethiourea formation at environmentally-relevant concentrations of ethylenethiourea in a pooled groundwater sample

N-nitroso compounds form from the interaction between nitrosatable precursors and nitrite under acidic conditions. A majority of N-nitroso compounds tested show evidence of carcinogenicity in animal models. Formation of N-nitroso compounds may occur from exposure to precursors in drinking water, but the extent of formation depends on a number of factors, including concentration of substrates, presence of catalysts and inhibitors, and pH. The objective of this study was to examine these factors in pesticide-associated N-nitroso (PANN) compound formation in drinking water. In preliminary screening experiments, nine nitrosatable pesticides and degradation products were individually reacted at environmentally-relevant concentrations (≤ 20 μg L‐−1) with sodium nitrite (NaNO2) and hydrochloric acid (HCl) in ultra-pure water. Only ethylenethiourea (ETU) showed evidence of PANN compound formation in initial experiments and was further tested for N-nitrosoethylenethiourea (N-ETU) formation in a pooled groundwater sample (comprised of five tap water samples combined into one homogenous sample) collected from an agricultural region of Prince Edward Island in Canada, where nitrate contamination is a known concern. Evidence of N-ETU formation in the groundwater sample was observed within 30 min at concentrations 7.5, 10, and 20 μg L−1. Analysis of target compounds and semi-target PANN compounds was performed using ultra-high pressure liquid chromatography coupled with high resolution orbital ion trap mass spectrometry. These preliminary experiments serve to inform about potential PANN compound formation in groundwater. The results of this study suggest that ETU is capable of forming potentially carcinogenic N-ETU in water containing nitrite/nitrate at trace concentrations under acidic conditions. Thus, these findings suggest that N-ETU formation may be a concern for individuals exposed to low concentrations of ETU in groundwater.

Comparison of sample preparation approaches and validation of an extraction method for nitrosatable pesticides and metabolites in human serum and urine analyzed by liquid chromatography – Orbital ion trap mass spectrometry

In the acidic environment of the stomach, nitrosatable pesticide residues may react with nitrite to form potentially carcinogenic pesticide-associated N-nitroso compounds (PANOCs). The objective of this study was to develop a method for the analysis of 10 nitrosatable pesticides and breakdown products in human serum and urine. Three sample preparation methods were evaluated for extraction of target analytes from the biomatrices. Deproteinization by methanol for 300-μL aliquots of serum with a final extract volume of 225 μL resulted in excessive ion enhancement of some analytes and suppression of others. Three types of solid-phase extraction cartridges were tested for optimal analyte retention from 200-μL aliquots of serum with a final extract volume of 400 μL; this approach resulted in significant analyte loss for some compounds. The Quick, Easy, Cheap, Effective, Rugged, and Safe approach resulted in a suitable method for extraction of the analytes from each biomatrix. Biofluid samples (500 μL) were spiked to 100 μg L−1 with analytical standards and extracted using 500 μL of acetonitrile (ACN) with 4% acetic acid (AcOH) for serum and 0.1% AcOH in ACN for urine. For extraction, 200 mg magnesium sulfate (MgSO4) and 50 mg sodium acetate were added for serum and 200 mg MgSO4 and 50 mg sodium chloride were added for urine. Final extract volumes for both biomatrices using the QuEChERS method was 400 μL after dilution. Samples were analyzed via ultra-high pressure liquid chromatography/high-resolution accurate mass orbital ion trap mass spectrometry. Mean recoveries for target analytes in serum and urine ranged between 74 and 120% (%RSD < 12) and 96 to 116% (%RSD ≤ 10), respectively. These methods may be used in large-scale biomonitoring studies to analyze PANNs and their parent compounds in human serum and urine.

Identification and Determination of Rubrofusarin, Rubrofusarin Isomer, and Their Quinone Forms in Grains Using High-Resolution Mass Spectrometry.

Fungi of the genus Fusarium can produce secondary metabolites such as naphthopyrones and naphthoquinones that are toxic and expected to threaten the food and feed safety. In this study, the occurrence of rubrofusarin, rubrofusarin isomer, and their quinone forms in grains was identified and confirmed using ultrahigh-performance liquid chromatography coupled with hybrid quadrupole orbital ion trap mass spectrometry (Q-Orbitrap MS). The quantitation of these compounds in grain samples was also investigated using Q-Orbitrap MS. The results showed the concentrations of rubrofusarin ranged from 3.278 to 33.82 μg/kg, from 0.815 to 61.86 μg/kg, and from 7.362 to 47.24 μg/kg for the maize, rice, and wheat samples, respectively. By comparison, the abundances of their quinone forms were relatively lower, and the concentration of quinone form of rubrofusarin isomer was relatively higher than that of quinone form of rubrofusarin. These compounds were also confirmed to coexist with other known Fusarium mycotoxins. The data-dependent tandem mass spectra obtained from the Q-Orbitrap MS were validated to provide a wealth of valuable information that allowed for advanced data interpretation for solid confirmation of these compounds in grains. To the best of our knowledge, this is the first study that concerns the occurrence and quantitation of rubrofusarin, rubrofusarin isomer, and their quinone forms in grains.

Single-Laboratory Validation Study of a Method for Screening and Identification of Phosphodiesterase Type 5 Inhibitors in Dietary Ingredients and Supplements Using Liquid Chromatography/Quadrupole-Orbital Ion Trap Mass Spectrometry: First Action 2015.12.

A single-laboratory validation study of a method for screening and identification of phosphodiesterase type 5 (PDE5) inhibitors in dietary ingredients and supplements is described. PDE5 inhibitors were extracted from the samples using a 50:50 (v/v) mixture of acetonitrile and water and centrifuged. Supernatant was diluted, filtered, and analyzed by LC-high-resolution MS. Data were collected in MS acquisition mode that combined full-scan MS experiment with all-ion fragmentation and data-dependent MS/MS product from the ion scan experiment. This approach enabled collection of MS and tandem MS (MS/MS) data for both targeted and nontargeted PDE5 inhibitors in a single chromatographic run. Software-facilitated identification of targeted analytes was performed based on the retention time, accurate mass, and isotopic pattern of pseudomolecular ions, and accurate masses of fragment ions using an in-house compound database. Detection and identification of other PDE5 inhibitors and novel analogs were performed by retrospective evaluation of MS and MS/MS experimental data. The method validation results obtained for evaluated matrixes fulfilled the probability of identification requirements and probability of detection requirements (for the pooled data) set at 90% (95% confidence interval) in the respective AOAC Standard Method Performance Requirements for identification and screening methods for PDE5 inhibitors. Limited data demonstrating the quantification capability of the method were also generated. Mean recovery and repeatability obtained for the evaluated PDE5 inhibitors were in the range 69-90% and 0.4-1.8%, respectively.

Identification of Metallothionein Subisoforms in HPLC Using Accurate Mass and Online Sequencing by Electrospray Hybrid Linear Ion Trap-Orbital Ion Trap Mass Spectrometry

A comprehensive approach to the characterization of metallothionein (MT) isoforms based on microbore HPLC with multimodal detection was developed. MTs were separated as Cd(7) complexes, detected by ICP MS and tentatively identified by molecular mass measured with 1-2 ppm accuracy using Orbital ion trap mass spectrometry. The identification was validated by accurate mass of the corresponding apo-MTs after postcolumn acidification and by their sequences acquired online by higher-energy collision dissociation MS/MS. The detection limits down to 10 fmol and 45 fmol could be obtained by ESI MS for apo- and Cd(7)-isoforms, respectively, and were lower than those obtained by ICP MS (100 fmol). The individual MT isoforms could be sequenced at levels as low as 200 fmol with the sequence coverage exceeding 90%. The approach was successfully applied to the identification of MT isoforms induced in a pig kidney cell line (LLC-PK(1)) exposed to CdS nanoparticles.