Source: EASTERN REGIONAL RES CENTER submitted to
DEVELOPMENT, EVALUATION, AND VALIDATION OF TECHNOLOGIES FOR THE DETECTION AND CHARACTERIZATION OF CHEMICAL CONTAMINANTS IN FOODS
Sponsoring Institution
Agricultural Research Service/USDA
Project Status
TERMINATED
Funding Source
Reporting Frequency
Annual
Accession No.
0430153
Grant No.
(N/A)
Project No.
8072-42000-080-000D
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jan 19, 2016
Project End Date
Jan 18, 2021
Grant Year
(N/A)
Project Director
LEHOTAY S J
Recipient Organization
EASTERN REGIONAL RES CENTER
(N/A)
WYNDMOOR,PA 19118
Performing Department
(N/A)
Non Technical Summary
(N/A)
Animal Health Component
(N/A)
Research Effort Categories
Basic
25%
Applied
75%
Developmental
0%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
1332410200018%
7113910200018%
7235010200018%
1335220200018%
7117299200028%
Goals / Objectives
1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats.
Project Methods
The specific approaches for meeting the project¿s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats.

Progress 01/19/16 to 01/18/21

Outputs
PROGRESS REPORT Objectives (from AD-416): 1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Approach (from AD-416): The specific approaches for meeting the project⿿s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. This is the final report for Project 8072-42000-080-00D, which ended January 18, 2021. New approved project 8072-42000-088-00D, entitled ⿿Technology Development, Evaluation, and Validation for the Detection and Characterization of Chemical Contaminants in Foods,⿝ has been established. Chemical residue monitoring of food is required in laboratories worldwide to ensure food safety, protect the environment, meet laws, and avoid liabilities. This project addressed the need to detect multiple chemical residues and other toxic compounds in foods most effectively and efficiently. The project met the needs of the USDA Food Safety Inspection Service (FSIS), Food and Drug Administration (FDA), and other organizations that monitor chemical residues in food, including industry, consumer groups, and academic scientists. Altogether, 63 peer-reviewed publications resulted from this project since 2016, and numerous abstracts, proceedings, lectures, and training courses. The ARS scientists developed and validated advantageous analytical methods which were transferred to and implemented by FSIS in the National Residue Program, including the ⿿extract & inject⿝ analytical method for veterinary drugs in catfish, ready-to-eat meats, liquid and powdered eggs, and cattle, chicken, and pork muscle, kidney, and liver. The newest ARS method, known as the ⿿quick, easy, cheap, effective, rugged, safe, efficient, and robust⿝ (QuEChERSER) mega-method has been extensively validated and is being transferred to FSIS to monitor for pesticides, veterinary drugs, and environmental contaminants in the same procedure. In this way, at least two methods are being combined into one method, which has the most impact on reducing the time, costs, and labor associated with analyses in the laboratory. The outcome of this research will be faster, simpler, and more accurate monitoring of pesticides, veterinary drugs, toxins, endocrine disruptors, and mercury and arsenic species in all types of foods. A noticeable impact of this research will be improved human health and more responsible food production practices through enhanced food safety due to better regulatory enforcement, more effective control of food trade, and higher quality of data for risk assessment and other purposes. Comparison of different mass spectrometric identification criteria for regulatory analysis of chemical residues in food. Chemical analysis requires both qualitative and quantitative information for a regulatory agency to make an enforcement action. Tandem mass spectrometry (MS/MS) is the most common analytical tool used for monitoring the food supply for toxic chemicals. Certain MS/MS criteria need to be met for qualitative identifications to be made, and several entities worldwide have devised different criteria for regulatory purposes. ARS scientists in Wyndmoor, Pennsylvania, compared the three most common sets of identification criteria in the validation of a simple ⿿extract & inject⿝ sample preparation method for 169 veterinary drugs in eggs. Although the current MS/MS identification criteria used by the USDA performed just as well as the other sets of criteria, one of the European criteria were simpler to implement. The USDA National Residue Program should adopt the simpler qualitative identification criteria evaluated in this study to improve monitoring efficiency and harmonize with global standards. Development of a contactless polytetrafluoroethylene membrane gas-liquid separator (GLS). Membrane GLS has problems with clogging, structural deterioration, and operation disruption. ARS scientists in Wyndmoor, Pennsylvania, developed a new GLS that eliminates liquid-membrane contact to greatly extend the ruggedness of the device. For elements with unstable hydrides, such as lead and cadmium, the small interior volume of the GLS also reduces residence time and sample degradation. This research will improve the analysis of toxic metals in foods, which helps to increase food safety. All objectives and milestones of the 5-year project were fully met, except for Objective 6, which was ended due to the resignation and abolishment of the position in 2018 of the scientist leading investigations on antimicrobial resistance. Analytical methods using mass spectrometry for the detection of biomarkers indicating antimicrobial resistance were developed, but the planned correlation with samples containing antibiotic residues could not be conducted. The COVID-19 pandemic and closure of the lab in March 2020 for more than 15 months limited the monitoring of samples for pesticides, veterinary drugs, and environmental contaminants, but the food packaging analyses were completed before the lab closure. ACCOMPLISHMENTS 01 Identifying chemicals migrating from food packaging. ARS scientists in Wyndmoor, Pennsylvania, in collaboration with USDA Food Safety and Inspection Service (FSIS), studied the migration of chemicals from paper and plastic food contact materials, including pizza boxes, butcher paper, egg containers, oven bags, and microwave and meal trays. A novel approach for the confident identification of chemicals using high-resolution mass spectrometry was developed. Over 100 migrated chemicals were identified, including substances intentionally added to food contact materials worldwide and their derivatives and degradation products. Concentrations of the identified chemicals with established migration limits were below the regulatory levels, suggesting no health concerns. However, some chemicals were reported for the first time, warranting future investigations.

Impacts
(N/A)

Publications

  • Sapozhnikova, Y.V. 2021. Non-targeted screening of chemicals migrating from paper-based food packaging by GC-Orbitrap mass spectrometry. Talanta. Volume 226: 1-10. https://doi.org/10.1016/j.talanta.2021.122120.
  • Michlig, N., Lehotay, S.J., Lightfield, A.R., Horacio, B., Maria, R. 2021. Validation of a high-throughput method for analysis of pesticide residues in hemp and hemp products. Journal of Chromatography A. 1645. https://doi. org/10.1016/j.chroma.2021.462097.
  • Lehotay, S.J., Lightfield, A.R. 2021. Comparison of four different multiclass, multiresidue sample preparation methods in the analysis of veterinary drugs in fish and other food matrices. Analytical and Bioanalytical Chemistry. 3223⿿3241. https://doi.org/10.1007/s00216-021- 03259-x.
  • Sapozhnikova, Y.V., Nunez, A., Johnston, J. 2021. Screening of chemicals migrating from plastic food contact materials for oven and microwave applications by LC- and GC-Orbitrap MS. Journal of Chromatography A. 462261. https://doi.org/10.1016/j.chroma.2021.462261.
  • Lehotay, S.J., De Zeeuw, J., Sapozhnikova, Y.V., Michling, N., Rousova, J., Konschnik, J.D. 2020. There is no time to waste: Vacuum gas chromatography ⿿ mass spectrometry is a proven low pressure (LP)GC-MS solution for fast, sensitive, and robust analysis. LC GC North America. 38(8):457-464. http://www.chromatographyonline.com/
  • Ninga, E., Sapozhnikova, Y.V., Lehotay, S.J., Lightfield, A.R., Monteiro, S. 2020. High-throughput mega-method for the analysis of pesticides, veterinary drugs, and environmental contaminants in catfish by UHPLC-MS/MS and robotic mini-SPE cleanup + LPGC-MS/MS. Journal of Agricultural and Food Chemistry. https://doi.org/10.1021/acs.jafc.0c00995.
  • Ji, M., Jixin, L., Xuefei, M., Chen, G., Chunsheng, L., Yongzhong, Q. 2020. A portable and field optical emission spectrometry coupled with microplasma trap for high sensitivity analysis of arsenic and antimony simultaneously. Talanta. available online:Talanta 218 (2020)121161. https:/ /doi.org/10.1016/j.talanta.2020.121161.
  • Sapozhnikova, Y.V., Zomer, P., Gerssen, A., Nunez, A., Mol, H.G. 2020. Evaluation of flow injection mass spectrometry approach for rapid screening of selected pesticides and mycotoxins in grain and animal feed samples. Food Control. 116. https://doi.org/10.1016/j.foodcont.2020.107323.
  • Rodriguez-Ramos, R., Lehotay, S.J., Michlig, N., Socas-Rodriguez, B., Rodriguez-Delgado, M.A. 2020. Critical review and re-assessment of analyte protectants in gas chromatography. Journal of Chromatography A. https:// doi.org/10.1016/j.chroma.2020.461596.


Progress 10/01/19 to 09/30/20

Outputs
Progress Report Objectives (from AD-416): 1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Approach (from AD-416): The specific approaches for meeting the project⿿s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Progress was made in all objectives, and the project is progressing according to schedule, except for Objective 6 which will be ended due to the resignation on 9/30/18 of the scientist leading investigations on antimicrobial resistance and abolishment of the position. Analytical methods using mass spectrometry for detection of biomarkers indicating antimicrobial resistance were developed, but the planned correlation with samples containing antibiotic residues could not be conducted. Otherwise, Objectives 1A, 1B, 2, 3, 4, and 5 are fully met. The covid-19 pandemic and closure of the lab in March 2020 for more than 3 months delayed the planned monitoring of samples for pesticides, veterinary drugs, and environmental contaminants. Fortunately, the food packaging leaching study had been completed before the lab closure. A gas chromatograph ⿿ orbital ion trap mass spectrometer has been installed through a material transfer research agreement. Development of a low-volume membrane gas-liquid separator (MGLS) for hydride generation (HG) in analysis of lead and cadmium. Simultaneous analysis of lead (Pb) and cadmium (Cd) in a variety of sample types, including foods, at the same time as other metals that require HG in their analysis, such as inorganic arsenic (iAs) is challenging proposition. ARS scientists in Wyndmoor, Pennsylvania, demonstrated the feasibility of HG for analysis of Pb and Cd by developing a MGLS using a 1.9 mm outer diameter, 1 µm porous polytetrafluoroethylene (PTFE) tubular membrane. The MGLS has a low intrinsic volume to minimize contact time and hence increases interaction between hydride products and sample solution. The design has an additional advantage to minimize polyatomic interferences, which is especially useful when inductively -coupled plasma ⿿ mass spectrometry (ICP-MS) is used for detection. The MGLS is a key step in the development of a multi-element method that includes Cd, Pb, and iAs in the same analysis of food samples. Accomplishments 01 Development of the ⿿quick, easy, cheap, effective, rugged, safe, efficient, and robust⿝ (QuEChERSER) mega-method to analyze pesticides, veterinary drugs, and environmental contaminants in foods. In 2003, the QuEChERS approach to sample preparation was introduced for pesticide residue analysis in foods, which has become the primary method used worldwide in the application, but instrumentation and technology has continued to improve in the past 17 years, creating a need to update the QuEChERS method. Using modern tools of mass spectrometry for detection, ARS scientists in Wyndmoor, Pennsylvania, expanded analytical scope to include veterinary drugs in the same method, further streamlined the steps, and added an automated cleanup technique conducted in parallel with analysis. So far, the new QuEChERSER mega- method has been validated for up to 349 diverse analytes in fish, bovine, caprine, and ovine muscle, hemp pruducts, and fruits and vegetables. QuEChERSER is expected to eventually supplant QuEChERS as the primary method for monitoring a wide array of chemical contaminants in foods. 02 Development of a fast screening method for the analysis of pesticides and mycotoxins in food and feed. Fast and simple screening methods are needed for the analysis of harmful contaminants in food and feed samples in regulatory testing. Currently, the fastest instrumental analysis takes at least 10 minutes by liquid chromatography-mass spectrometry. ARS scientists in Wyndmoor, Pennsylvania, in collaboration with scientists from Wageningen Food Research in the Netherlands, developed a 2-minute analysis using flow injection coupled with mass spectrometry for fast screening of 12 pesticides and 7 mycotoxins in complex samples of grains and animal feed. The developed approach allows rapid screening of pesticides and mycotoxins at their respective U.S. tolerances and the European Union (EU) maximum residue limits. Compared to existing methods, the new method provides simplicity, higher throughput, lower cost, and efficient use of high- end instrumentation. 03 Accumulation of contaminants in fish influenced by snowmelt and municipal effluent discharge. ARS scientists in Wyndmoor, Pennsylvania, in collaboration with scientists from Baylor University in Waco, Texas, studied the accumulation of pesticides and environmental contaminants in fish collected from East Canyon Creek, Utah. The creek is in a semi- arid ecosystem fed by snowmelt in spring and municipal effluent discharge in the summer and fall. Fish samples (brown trout and mottled sculpin) collected in four seasons at incremental distances downstream of the effluent discharge showed accumulation of 18 contaminants at low ng/g levels. The seasons did not make a difference. The highest levels of certain banned flame retardants (polybrominated diphenyl ethers) were found in fish collected close to the effluent discharge.

Impacts
(N/A)

Publications

  • Fialkov, A.B., Lehotay, S.J., Amirav, A. 2019. Less than 1 minute low- pressure gas chromatography - mass spectrometry. Journal of Chromatography A. 1612 (2020) 460691.
  • Lehotay, S.J., Lightfield, A.R. 2020. Extract-and-inject analysis of veterinary drug residues in catfish and ready-to-eat meats by ultrahigh- performance liquid chromatography ⿿ tandem mass spectrometry. Journal of AOAC International. 103(2): 584-606.
  • Lehotay, S.J., Michlig, N., Lightfield, A.R. 2020. Assessment of test portion sizes after sample comminution with liquid nitrogen in the high- throughput analysis of pesticide residues in fruits and vegetables. Journal of Agricultural and Food Chemistry. 68:1468-1479.
  • Chen, G., Lai, B., Chen, T., Mao, X. 2020. Brief soaking at above- gelatinization temperature reduces inorganic arsenic in cooked rice. Cereal Chemistry.
  • Monteiro, S.H., Lehotay, S.J., Sapozhnikova, Y.V., Ninga, E., Lightfield, A.R. 2020. High-throughput mega-method for the analysis of pesticides, veterinary drugs, and environmental contaminants in beef by UHPLC-MS/MS and robotic mini-SPE cleanup + LPGC-MS/MS. Journal of Agricultural and Food Chemistry.
  • Sapozhnikova, Y.V., Salamova, A., Haddad, S., Burket, S.R., Luers, M., Brooks, B. 2020. Spatial and seasonal occurrence of pesticides and environmental contaminants in fish tissues influenced by snowmelt and municipal effluent discharge. Science of the Total Environment.
  • Roussey, M., Lehotay, S.J., Pollaehne, J. 2019. Cryogenic Sample Processing with Liquid Nitrogen for Effective and Efficient Monitoring of Pesticide Residues in Foods and Feeds. Journal of Agricultural and Food Chemistry. 67: 9203-9209.


Progress 10/01/18 to 09/30/19

Outputs
Progress Report Objectives (from AD-416): 1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Approach (from AD-416): The specific approaches for meeting the project⿿s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Progress was made in all objectives, and the project is progressing according to schedule, except for Objective 6 which will be delayed due to the resignation on 9/30/18 of the scientist leading investigations on antimicrobial resistance. A visiting scientist will begin on 9/1/19 for one year to contribute to Objective 6, and recruitment to fill the vacancy will commence when permitted. Otherwise, Objectives 1A, 2, 3, 4, and 5A are essentially complete. A method for >300 pesticides, veterinary drugs, and environmental contaminants has been validated in food animal tissues using FSIS standards, including qualitative identification involving blind analyses. Due to the purchase of new instrumentation, Objective 5B will use cold trapping ⿿ hydride generation ⿿ inductively-coupled plasma ⿿ mass spectrometry instead of or in addition to the originally planned graphite furnace - atomic fluorescence spectroscopy for analysis of inorganic arsenic. The food packaging study in Objective 1B has been initiated and new instrumentation (gas chromatograph ⿿ orbital ion trap mass spectrometer) has been obtained through a material transfer research agreement to conduct nontargeted analysis of food packaging components in addition to targeted analysis as originally planned. The validation and monitoring aspects of Objectives 1B and 1C will be completed as planned in the final years of the project. Accomplishments 01 Reduction of inorganic arsenic (iAs) concentration during cooking of rice. Rice is the staple food for half of the world⿿s population, but it also contains a much higher concentration of iAs, a Class-1 carcinogen, than other grains or vegetables. ARS researchers at Wyndmoor, Pennsylvania, developed an effective procedure to reduce iAs levels by first soaking 1-part rice at 80 degrees C in 10 parts water, which is discarded after 10 minutes, and then cooking the rice in 2 parts water as usual in East Asia. On average, a 40% reduction in iAs concentration was achieved by this method due to the higher solubility of iAs in the hot water that also contains the starchy gelatinous components from the rice. This pre-soaking method is easily implemented by the public to reduce chronic arsenic exposure using common cooking practices. Those who follow this simple approach will cut their risk of cancer, which could impact billions of people worldwide if this information is widely disseminated. 02 Development of hydride generation - cryogenic trapping - inductively- coupled plasma - mass spectrometry (HG-CT-ICP-MS) for analysis of inorganic arsenic (iAs). ICP-MS is the benchmark technique for elemental analysis due to its high sensitivity and wide dynamic range, but the toxic iAs species must first be separated from the less toxic metalorganic forms of arsenic prior to analysis of samples. Conventional methods of iAs separation are slow and use expensive equipment and large amounts of reagents. Cryogenic trapping advantageously uses temperature rather than chemical reagents to speciate arsenic due to boiling point differences. ARS researchers at Wyndmoor, Pennsylvania, developed HG-CT-ICP-MS for the first time in analytical chemistry to analyze iAs in different foods. This environmentally green system reduces costs by requiring less instrumentation and reagents, including argon gas for ICP-MS, and greatly increases sample throughput. The cryotrapping device for this approach has been patented by ARS and is available for possible commercialization and widespread implementation. 03 Development and validation of a method for analysis of >300 pesticides and environmental contaminants in catfish. Catfish is one of the most consumed freshwater fish in the USA, valued for its nutritional benefits, but it is a bottom feeder, which causes concerns about potential accumulation of chemical contaminants. ARS scientists at Wyndmoor, Pennsylvania, developed and validated a simple, fast and efficient analytical method for the analysis of 302 pesticides and environmental contaminants in catfish muscle. The method was successfully validated at and below U.S. regulatory levels of concern and applied to the analysis of fish from the market. The method is being implemented by the USDA Food Safety and Inspection Service (FSIS) for routine monitoring of contaminants in catfish in the U.S. National Residue Program. The new method provides high sample throughput, wide monitoring scope and reduced cost of analysis.

Impacts
(N/A)

Publications

  • Perez Jr, J.J., Chen, C. 2018. Detection of acetyltransferase modification of kanamycin, an aminoglycoside antibiotic, in bacteria using ultra-high performance liquid chromatography tandem mass spectrometry. Journal of Rapid Communications in Mass Spectroscopy. 32:1549-1556.
  • Perez Jr, J.J., Chen, C. 2018. Rapid detection and quantification of aminoglycoside phosphorylation products using direct infusion high resolution and ultra-high performance liquid chromatography-mass spectrometry. Rapid Communications in Mass Spectrometry. 32:1822-1828.
  • Sapozhnikova, Y.V. 2018. Development and validation of a semi-automated high-throughput analytical method for 265 pesticides and environmental contaminants in meats and poultry. Journal of Chromatography A. 1572:203- 211.
  • Chen, G., Lai, B., Mei, N. 2018. Open-vessel digestion of fish muscle with minimal analyte loss in mercury speciation analysis. Talanta. 191:209-215.
  • Chen, T., Lu, J., Kang, B., Lin, M., Ding, L., Zhang, L., Chen, G., Chen, S., Lin, H. 2018. Antifungal activity and action mechanism of ginger oleoresin against Pestalotiopsis Microspora isolated from Chinese olive fruits. Frontiers in Microbiology.
  • Lehotay, S.J. 2018. Possibilities and limitations of isocratic fast liquid chromatography-tandem mass spectrometry analysis of pesticide residues in fruits and vegetables. Chromatographia. 82:235-250.
  • Lehotay, S.J., Han, L., Sapozhnikova, Y.V. 2018. Use of a quality control approach to assess measurement uncertainty in the comparison of sample processing techniques in the analysis of pesticide residues in fruits and vegetables. Analytical and Bioanalytical Chemistry. 1-15.
  • Chaney, R.L., Green, C.E., Lehotay, S.J. 2018. Inter-laboratory validation of an inexpensive streamlined method to measure inorganic arsenic in rice grain. Analytical and Bioanalytical Chemistry.
  • Lehotay, S.J., Chen, Y. 2018. Hits and misses in research trends to monitor contaminants in foods. Analytical and Bioanalytical Chemistry. 410:5331-5351.
  • Liu, M., Ding, L., Liu, J., Mao, X., Na, X., Chen, G., Qian, Y. 2019. Determination of arsenic in biological samples by slurry sampling hydride generation atomic fluorescence spectrometry using in-situ dielectric barrier discharge trap. Journal of Analytical Atomic Spectrometry.
  • Sapozhnikova, Y.V., Hoh, E. 2019. Suspect screening of chemicals in food packaging plastic film by comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry. LC GC North America. 37:52-65.
  • Nunez, A., Sapozhnikova, Y.V., Lehotay, S.J. 2018. Characterization of MS/ MS product ions for the differentiation of structural isomeric pesticides by high-resolution mass spectrometry. Toxics. 6(4):59. -.
  • Perez Jr, J.J., Chen, C. 2018. Implementation of normalized retention time (IRT) for bottom-up proteomic analysis of the aminoglycoside phosphotransferase enzyme facilitating method distribution. Analytical and Bioanalytical Chemistry. 411:4701-4708.


Progress 10/01/17 to 09/30/18

Outputs
Progress Report Objectives (from AD-416): 1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Approach (from AD-416): The specific approaches for meeting the project�s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Progress was made in all objectives, and the project is progressing according to schedule, and even ahead of schedule in objectives 2, 3, and 4. Objectives 1A and 3A are complete, and for 3B, cryogenic sample processing equipment and techniques were evaluated and compared with room temperature approaches for different food commodities and test portion sizes. The room temperature method and devices are sufficient to meet regulatory needs with smaller sample sizes than commonly used now, which enables further reduction in reagent costs and substantially higher sample throughput. With respect to Objective 5, novel analysis to speciate mercury in foods was achieved by designing a method using a quartz-coil reactor to perform differential photochemical vapor generation following by atomic fluorescence spectroscopy. For analysis of inorganic arsenic, a new approach using a thermoelectric cryogenic trap was successfully validated using rice flour certified reference material. On Objective 6, multiple methods for rapid determination of an aminoglycoside antibiotic modification by resistant bacteria were developed. In addition, a bottom-up proteomics approach combined with computational prediction technologies for targeted antibiotic resistance proteins was developed. Lastly, alternative commercial proteases were explored using antimicrobial resistant bacteria to validate cleavage efficiency. Accomplishments 01 Method for analysis of pesticides and environmental contaminants in meats and poultry. The USDA Food Safety and Inspection Service (FSIS) is responsible for routine monitoring of pesticide residues in meat and poultry products to assure that regulatory tolerances are not exceeded. ARS scientists at Wyndmoor, Pennsylvania developed and validated a fast, efficient high-throughput analytical method for FSIS-priority pesticides and environmental contaminants in meat (swine, cattle) and poultry (chicken) muscle. Satisfactory validation results were achieved for 219 contaminants. The method was transferred to FSIS laboratories, and is currently being implemented for routine monitoring of contaminants. The implementation of the new method is expected to improve regulatory monitoring, providing reduced cost per sample analysis, increased sample throughput, and reliable data for more contaminants of concern. 02 Reduction of inorganic arsenic content in cooked rice. Rice is a staple food for half the world population, but it contains higher levels of toxic inorganic arsenic (iAs) than other common crops. Chronic exposure to iAs through dietary pathway is a worldwide concern for rice consumers. ARS researchers at Wyndmoor, Pennsylvania developed an effective protocol to reduce iAs in cooked rice by presoaking the rice in hot water for 10 minutes, then discarding the water before cooking until the rice becomes dry using a rice cooker. Previous efforts to reduce iAs in cooked rice took longer and/or were less effective. This more rapid protocol achieved similar or better iAs reduction by raising soaking temperature above the gelatinization temperature of rice starch, which causes higher diffusion kinetics to reduce iAs levels. Implementation of this protocol in daily practice would cut cancer risk of rice consumers and improve their long-term health prospects. 03 Proteomic analysis of computationally predicted peptides from antimicrobial resistant bacteria genomes. As antimicrobial resistance continues to become a global issue, better and faster analytical methods need to be devised to slow its progression. Proteomics could be a viable option, but little research has been conducted to fulfill this purpose because large scale proteomics experiments are costly. Instead, open source software platforms are available to streamline proteomics studies without need for extensive empirical measurements. ARS scientists at Wyndmoor, Pennsylvania tested this concept for targeted analysis of enzymes associated with antibiotic resistance using only its genome to predict the tryptic digest peptides generated. This technique facilitates subsequent method development using liquid chromatography - mass spectrometry for implementation. In a feasibility study, comparable digestion efficiencies for two proteases determined by this approach suggests that the more cost-effective method can be used for large scale proteomic investigations in the future. 04 Streamlined extraction for analysis of mercury in seafoods. Traditional open-vessel extraction of mercury from foods for analysis suffers from analyte loss via volatilization, but ARS researchers at Wyndmoor, Pennsylvania designed a better reflux digestion system that consisted of a block heater and multiple Pyrex culture tubes to provide improved extraction efficiency. Unlike closed-vessel systems, this open- vessel setup allows easy dissipation of nitrous oxides and nitrites that normally interfere in the analysis. This protocol was validated using a certified reference material, achieving quantitative recovery with <5% relative errors. This approach only costs $1,000 for materials compared to $35,000 for a traditional closed-vessel microwave-assisted extraction system, including the vessels, and provides equivalent results and potential greater sample throughput. 05 A new analytical method for organophosphate ester plasticizers and flame retardants in meats and fish. ARS scientists at Wyndmoor, Pennsylvania, developed a new method for analysis of 14 organophosphate ester (OPE) plasticizers and flame retardants in meats and fish. OPEs are widely used as additives to plastics, electronics, furniture and textiles, and easily migrate from materials to the environment. Some OPEs are persistent, bioaccumulative, and toxic. The developed method was based on the QuEChERS approach and automated robotic cleanup of the extracts. The final extracts were split and analyzed in parallel by gas and liquid chromatography coupled to tandem mass spectrometry to provide an additional degree of confidence in the results. The new method was successfully applied to the analysis of real-world meat and fish samples form the market, further demonstrating the utility of the method for implementation in regulatory and commercial laboratories. 06 Rapid determination of antimicrobial resistance of aminoglycoside antibiotics. Microbial resistance to antibiotic drugs, such as aminoglycosides, is a major health concern worldwide, and more efficient and effective methods of analysis are needed to monitor and study resistance. ARS researchers in Wyndmoor, Pennsylvania have developed an analytical method capable of determining antibiotic resistance via acetylation and phosphorylation in less than 3 hours following overnight enrichment. This is important because aminoglycosides and their enzyme-modified form are challenging to analyze using conventional HPLC instrumentation. These methods allow their detection/quantification and provides the framework for an all- inclusive antibiotic modification evaluation method. In-house developed software was then used for rapid quantification of the biomarkers. 07 Identification of food packaging chemicals extracted from plastic stretch film. Food packaging is important in protecting food, extending its shelf-life, and providing consumers with food handling convenience. However, during storage and handling, some chemicals from food packaging materials may potentially migrate into packaged food. ARS scientists at Wyndmoor, Pennsylvania, in collaboration with San Diego State University scientists, identified chemicals extracted from food packaging plastic stretch film using comprehensive two-dimensional gas chromatography coupled to time-of-flight mass spectrometry (GC�GC/ TOF-MS). Common plasticizers, polymer and plastic additives, UV filters, fragrances, flavoring agents, among others, were tentatively identified. These data offer an insight on identity of potential food packaging contaminants, and pave the way for future studies on migration of food packaging chemicals into real foods needed for risk assessment to protect consumer�s health. 08 Determination of contaminants in mussels and oysters from Hong Kong. Aquaculture operations are growing worldwide and will continue to increase to meet global food demands. ARS scientists at Wyndmoor, Pennsylvania, in collaboration with researchers from Baylor University, Texas, and Hong Kong Baptist University, Hong Kong, examined bioaccumulation of contaminants by marine bivalves located near sewage and landfill wastewater discharges in Hong Kong, the fourth most densely populated country in the world. Multiple classes of pharmaceutical residues, pesticides, polycyclic aromatic hydrocarbons (PAHs), and flame retardants were detected at low ng/g levels. Initial estimates of acceptable servings per week indicated low consumption risks for measured contaminants with developed U.S. EPA reference dose. These data shed light on aquaculture product safety, particularly, the products from rapidly urbanizing regions of developing countries with limited infrastructure.

Impacts
(N/A)

Publications

  • Lehotay, S.J., Lightfield, A.R. 2018. Simultaneous analysis of aminoglycosides with many other classes of drug residues in bovine tissues by ultra high-performance liquid chromatography-tandem mass spectrometry using an ion-pairing reagent added to final extracts. Analytical and Bioanalytical Chemistry. 410:195-1109.
  • Han, L., Lehotay, S.J., Sapozhnikova, Y.V. 2017. Comparison of room temperature and cryogenic sample processing in the analysis of chemical contaminants in foods. Journal of Agricultural and Food Chemistry. 66:4986- 4996.
  • Song, S., Zhang, Z., Pan, C., Han, L., Sapozhnikova, Y.V. 2017. Determination of six parabens residues in fresh-cut vegetables using QuEChERS with multi-walled carbon nanotubes and high performance liquid chromatography-tandem mass spectrometry. Journal of Food Analytical Methods. 10:3972-3979.
  • Burket, S., Sapozhnikova, Y.V., Zheng, J., Chung, S., Brooks, B.W. 2018. At the intersection of urbanization, water and food security: bioaccumulation of select contaminants of emerging concern in mussels and oysters from Hong Kong. Journal of Agricultural and Food Chemistry. 66:5009-5017.
  • Song, S., Zhu, K., Han, L., Sapozhnikova, Y.V., Zhang, Z., Yao, W. 2018. Residue analysis of sixty pesticides in red swamp crayfish using QuEChERS with high-performance liquid chromatography-tandem mass spectrometry. Journal of Agricultural and Food Chemistry. 66:5031-5038.
  • Codling, E.E., Chen, G. 2017. Effects of compost amended lead-arsenate contaminated soils on total and inorganic arsenic concentration in rice. Journal of Plant Nutrition. 40(15):2146-2155.
  • Chen, G., Lai, B., Mei, N., Liu, J., Mao, X. 2017. Mercury speciation by differential photochemical vapor generation at UV-B vs. UV-C wavelength. Spectrochimica Acta B.
  • Chen, G., Lai, B. 2017. Determination of inorganic arsenic by hydride generation, thermoelectric cryogenic focusing, and atomic fluorescence spectrometry. Analytical Chemistry. 89:8678-8682.
  • Chen, G., Mei, N., Lai, B. 2017. Speciation of mercury in fish by photochemical vapor generation-atomic fluorescence spectrometry. Spectrochimica Acta B. 137:1-7.
  • Qi, Y., Mao, X., Liu, J., Na, X., Chen, G., Liu, M., Zheng, C., Qian, Y. 2018. An in-situ dielectric barrier discharge trap for ultrasensitive arsenic determination by atomic fluorescence spectrometry. Analytical Chemistry. 90:6332-6338.


Progress 10/01/16 to 09/30/17

Outputs
Progress Report Objectives (from AD-416): 1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Approach (from AD-416): The specific approaches for meeting the project�s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Progress was made on all objectives, all of which fall under National Program 108 � Food Safety, Component I, Foodborne Contaminants. Progress on this project focuses on Problem Statement D - Chemical and Biological Contaminants: Detection and Characterization Methodology, Toxicology, and Toxinology. Progress was made in all objectives, and the project is progressing according to schedule, and even ahead of schedule on Objective 3. A Research Chemist was hired in Sept. 2016 to meet Objective 6, which is intended to meet needs for better methods of analysis for antimicrobial resistance research using mass spectrometry for detection. To help meet this objective, a new Orbitrap tandem hybrid high resolution mass spectrometer was purchased and installed in Dec. 2016 for use Center-wide. Also, an inductively-coupled plasma � mass spectrometer (ICP-MS) was purchased and installed in March, 2017 for use by one of the investigators to help meet Objective 5 pertaining to the analysis of toxic elements in foods. Additionally, a gas chromatograph � triple quadrupole mass spectrometry (GC-MS/MS) instrument was recently upgraded in the lab to achieve state-of-the-art sensitivity for the detection of pesticides and other contaminants in foods. Furthermore, a robotic liquid autosampler was installed with this same instrument, including new software, which enables high-throughput analysis of >200 targeted chemicals in <13 min for each sample including automated cleanup. These powerful analytical tools have been used to develop automated streamlined high-throughput analytical methods, which also entail reliable data processing for hundreds of pesticides, veterinary drugs, and environmental and other contaminants with minimal human review and manual corrections. Improved analytical methods developed by the ARS scientists have already been transferred to USDA Food Safety and Inspection Service (FSIS), and similar technology transfer activities of this research project will continue with FSIS and other stakeholders as the even more efficient, sensitive, and accurate methods with wider scope are being developed and validated. Accomplishments 01 Evaluation and use of more reliable and efficient data processing for chemical analysis by chromatography. As analytical technologies and techniques have improved, sample throughput in chromatography has become limited by data processing of large generated data sets. Human review and manual corrections of integrated chromatographic peaks has been standard practice for decades, but this is no longer possible in high-throughput applications involving many targeted chemicals. ARS researchers at Wyndmoor, Pennsylvania empirically applied and evaluated a simple automated summation chromatographic peak integration approach for pesticide residue monitoring, which obviates the need for time- consuming human review and manual reintegrations. Improved results were achieved using summation integration in a much more efficient and reliable approach, which is applicable to chromatographic analysis in many fields of investigation. This simple, reliable, and automatic data handling approach for chromatography has been transferred to monitoring labs in the USDA-Food Safety and Inspection Service, and it is expected to be more widely implemented in the future. 02 Validation and transfer of an analytical method for veterinary drug residues in liquid and powdered eggs. The USDA-Food Safety and Inspection Service (FSIS) is responsible for monitoring the safety of liquid and powdered egg products, including analysis of veterinary drug residues to assure that regulatory tolerances are not exceeded. However, FSIS lacked a validated analytical method that could be used to determine the presence of the veterinary drug residues. At the request of FSIS, ARS researchers at Wyndmoor, Pennsylvania extended and validated their analytical method for food animal tissues to egg products. The results met FSIS validation criteria for >150 drug residues in the samples, and the method was transferred to FSIS for routine use in their monitoring laboratories. 03 Development of a cold-finger digestion system for speciation analysis of mercury (Hg) in fish. Traditional open-vessel sample digestion for metals analysis suffers from loss of relatively volatile elements, such as Hg. Closed-vessel microwave-aided digestion (MAD) partially overcomes this problem, but the expensive Teflon vessels compatible for microwave ovens can only accommodate a limited number of samples. To avoid MAD and Teflon vessels altogether, ARS researchers at Wyndmoor, Pennsylvania designed a cold-finger digestion system using 16 mm diameter x 150 mm tall Pyrex culture tubes placed in block heater. A temperature gradient in the tube headspace enables solvent reflux and condensation, thus minimizing analyte loss. The new approach and protocol obviates the need for additional chemical separation prior to analysis, saves thousands of dollars in equipment and supplies, and increases sample throughput, while still maintaining good analytical performance. 04 Development of an analytical technique for rapid determination of antimicrobial resistance of aminoglycoside antibiotics. Microbial resistance to antibiotic drugs, such as aminoglycosides, is a major health concern worldwide, and more efficient and effective methods of analysis are needed to monitor and study resistance. ARS researchers at Wyndmoor, Pennsylvania initiated development of a new method of detection by introducing into host bacteria plasmid encoding enzymes that modify aminoglycosides through acetylation and phosphorylation. After exposing the bacteria to aqueous aminoglycoside solution, mass spectrometric analysis revealed biomarkers useful for rapid detection of the resistant strain. In-house devised software was then used for rapid quantification of the biomarkers. This type of instrument-based approach using mass spectrometry possesses advantages over traditional microbiological methods, including complementary value. 05 Optimization of sample processing techniques to determine minimum test sample size for analysis of contaminants in foods. The best way to improve the efficiency in chemical residue analysis of foods is to use the minimum amount of test sample portion that still yields accurate results for the bulk sample. ARS researchers at Wyndmoor, Pennsylvania conducted a study involving 10 food commodities and two types of modern commercial sample processing devices to minimize test portions for analysis of added and incurred pesticide residues in the bulk samples. The results indicated that 5 g test samples processed at room temperature from 1 kg typically meets needs for regulatory analysis. This is half the portion commonly analyzed currently, which means that half the amount of reagents may be used thereby saving costs in the future. 06 Survey for the presence of mercury (Hg) in 38 fish oil supplement samples. Fish consumption is the main pathway of human exposure to inorganic mercury (iHg) and methylmercury (MeHg), and ARS researchers at Wyndmoor, Pennsylvania analyzed for these toxic chemicals in fish oil using a novel differential photochemical vapor generation atomic fluorescence spectrometric (PVG-AFS) method. Average concentrations of iHg and MeHg in the 38 fish oil samples were 0.67 and 1.1 ng/mL, respectively, which is 2-3 orders of magnitude lower than in fish. This study helped provide the fish oil industry insights and guidance for quality control and assurance of their products, and should increase consumer confidence in consumption of fish oil supplements. 07 Evaluation of a new sorbent material for efficient cleanup of complex food samples for the analysis of pesticides and environmental contaminants. Chemical analysis of residual contaminants in highly complex pigmented and fatty foods requires efficient cleanup of the food extracts to remove interfering matrix components. ARS researchers at Wyndmoor, Pennsylvania evaluated a new commercial sorbent, called Verde, which combines fat and pigment removing properties, in fatty and/ or pigmented commodities (avocado, pork, salmon, and pork) for analysis of 117 pesticides and environmental contaminants. The new sorbent provided efficient removal of chlorophyll and lipids, which enabled acceptable analysis of nearly all of the selected contaminants at or below regulatory tolerance levels. The new method is fast, efficient, and can be implemented in any laboratory for residual analysis of contaminants in foods. 08 Development of an extraction method for inorganic arsenic in algae. Algae are rich in innocuous arsenosugars but some algae species contain high levels of extremely toxic inorganic arsenic (iAs). ARS researchers at Wyndmoor, Pennsylvania in collaboration with the Institute of Quality Standards and Technology for Agri-Products, Chinese Academy of Agricultural Science, Beijing, China developed an improved effective method of analysis of iAs in algae. The new method eliminated interferences from arsenosugars and other organic arsenicals, achieving quantitative recovery and low detection limit of 3 ng/g for iAs. The method was applied in the analysis of real samples.

Impacts
(N/A)

Publications

  • Zhang, W., Qi, Y., Qin, D., Liu, J., Mao, X., Chen, G., Wei, C., Qian, Y. 2017. Determination of inorganic arsenic in algae using bromine halogenation and on-line nonpolar solid phase extraction followed by hydride generation atomic flourescence spectrometry. Talanta. 170:152-157.
  • Anumol, T., Lehotay, S.J., Stevens, J., Zweigenbaum, J. 2017. Comparison of veterinary drug residue results in animal tissues by ultrahigh- performance liquid chromatography coupled to triple quadrupole ... use of a commercial lipid removal product. Analytical and Bioanalytical Chemistry. 409:2639-2653.
  • Perez Jr, J.J., Watson, D.A., Levis, R.J. 2016. The classification of gunshot residue using laser electrospray mass spectrometry and offline multivariate statistical analysis. Analytical Chemistry. doi: 10.1021/acs. analchem.6b01438.
  • Han, L., Sapozhnikova, Y.V., Lehotay, S.J. 2016. Method validation for 243 pesticides and environmental contaminants in meats and poultry by tandem mass spectrometry coupled to low-pressure gas chromatography and ultra high-performance liquid chromatography. Food Control. 66:270-282.
  • Han, L., Sapozhnikova, Y.V., Matarrita, J. 2016. Evaluation of a new carbon/zirconia-based sorbent for the cleanup of food extracts in multiclass analysis of pesticides and environmental contaminants. Journal of Separation Science. 39(23):4592-4602.
  • Lehotay, S.J. 2017. Utility of the summation chromatographic peak integration function to avoid manual reintegrations in the analysis of targeted analytes. LC GC North America. 35:391-402.
  • Zhang, Z., Feng, M., Zhu, K., Han, L., Sapozhnikova, Y.V., Lehotay, S.J. 2016. Multiresidue analysis of pesticides in straw roughage by liquid chromatography - tandem mass spectrometry. Journal of Agricultural and Food Chemistry. 64:6091-6099. doi: 10.1021/acs.jafc.5b05981.


Progress 10/01/15 to 09/30/16

Outputs
Progress Report Objectives (from AD-416): 1. Develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants. 1A. Simultaneous analysis method for diverse pesticides, legacy and emerging environmental contaminants in meats. 1B. Multiresidue method for food packaging (FP) contaminants in packaged foods. 1C. Conduct a survey of food samples for emerging environmental and FP contaminants. 2. Develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives. 3. Develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation. 3A. Simultaneous analysis method for diverse veterinary drugs residues, including aminoglycoside antibiotics, in meats. 3B. Evaluate cryogenic sample processing to achieve meaningful representative sample size of 100 mg in multiresidue analysis of pesticides and veterinary drug residues in foods. 4. Develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods. 5. Develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods. 5A. Develop novel analytical methods for mercury speciation and quantification in foods. 5B. Develop novel analytical methods for arsenic speciation and quantification in foods. 6. Develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. Approach (from AD-416): The specific approaches for meeting the project�s objectives and milestones are as follows: 1) develop, validate, and transfer multiclass, multiresidue methods for pesticides, environmental, and emerging contaminants in FSIS-regulated foods, and conduct a survey of food samples for the contaminants; 2) develop qualitative screening and identification criteria with automated data processing that meets regulatory needs to minimize/avoid false positives and negatives; 3) develop sample processing methods for regulatory analysis that improve the ability to detect combinations of veterinary drug residues in the same sample preparation; 4) develop automated high-throughput sample processing, preparation, and analysis methods using flow-injection and/or open probe techniques coupled with mass spectrometry to monitor >500 veterinary drugs, pesticides, and environmental contaminants in foods; 5) develop novel analytical methods for inorganic and organometallic heavy metals [for example forms of mercury (Hg) and arsenic (As)] in foods; and 6) develop and use bioanalytical methods (including mass spectrometry) to monitor for antibiotic resistant organisms and/or their biomarkers in conjunction with antibiotic residues in seafood and meats. This new project plan was recently certified through the ARS Office of Scientific Quality Review (OSQR). For further details on current work see the 2016 annual report for project 8072-42000-056-00D.

Impacts
(N/A)

Publications

  • Lehotay, S.J., Han, L., Sapozhnikova, Y.V. 2016. Automated mini-column solid-phase extraction cleanup for high-throughput analysis of chemical contaminants in foods by low-pressure gas chromatography � tandem mass spectrometry. Chromatographia. 79:1113-1130. doi: 10.1007/s10337-016-3116- y.