Progress 05/01/12 to 04/30/16

Outputs
Target Audience:The target audience for this project are the scientists and regulators whom are interested in the protection of the environment and the food supply. During this period this audience was reached during presentations listed in the dissemination secton. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Dr. Brian Eitzer attended a seminar on the use of mass spectrometry in forensics at the State of Connecticut on January 20 How have the results been disseminated to communities of interest?Dr. Brian Eitzer presented a talk on the analysis of mycotoxins during the FDA FERN cCAP annual technical meeting in New Orleans on (March 28-31)(60 participants). Dr. BRIAN EITZER was an instructor for the class entitled "LB511 FDA/FERN Chemistry Training for LC/MS" which was held at the Food and Drug Administration's Forensic Chemistry Center in Cincinnati, OH from April 25-29 (12 students) What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? The use of HRMS to quantify pesticide residues in produce and environmental samples was validated as part of the laboratories ISO accreditation. This initial validation was conducted using standards spiked into a tomato matrix and then analyzed after a full QuEChERS based extraction and cleanup. All samples were then analyzed on the Thermo Exactive using a 3 scan function technique. Quantitation was done using the full scan HRMS while confirmation of the pesticide residue was performed using an all ion fragmentation scan. Two different fragmentation energies were used to increase the possible fragments to monitor. A pesticide had to have both a full scan HRMS parent and an HRMS fragment ion to be positively identified and confirmed. A total of 39 pesticides were found to be valid during this initial test, all with reporting limits below 10 PPB using this technique. This procedure is now part of our standard operating procedures for these analyses. We have also been using Tracefinder EFS as a screening tool for the analysis of pesticide residues in produce. This software can evaluate a much larger database of pesticides that have the potential to be found in produce samples. Pesticides that are identified using this technique will be added to the next round of method validation to determine if they can be reliably quantified at the levels necessary for reporting in produce. A second focus during this past year has been testing of an alternate HRMS instrument, a Thermo Q-Exactive. This instrument incorporates a quadrupole ion filter before the orbitrap. This additional filter allows more modes of analysis than our current Exactive. The older instrument can only do all ion fragmentation, while the newer version can isolate single ions, or ion ranges to perform true MS-MS. We explored the use of these alternate modes of analysis in a project in which we are analyzied a set of 12 mycotoxins in corn, wheat and peanut butter.

Publications

Progress 10/01/14 to 09/30/15

Outputs
Target Audience:The primary target audience for this project are regulators and other scientists whom have an interest in the protection of the environment and the food supply. During this period, the target audience reached were principally other scientists present at the presentations listed under dissemination. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Dr. Brian Eitzer attended the FDA FVM Science and Research Conference in Silver Springs, MD on (August 13-14). Dr. Brian Eitzer attended a workshop on the use of Tracefinder Software at the Thermo Instruments facility in Cambridge MA (September 14). Dr. Brian Eitzer attended training from Thermo Instruments on the use of the Q-Exactive at CAES on October 13-15. How have the results been disseminated to communities of interest?Dr. Brian Eitzer gave a presentation entitled "Determination of Pesticide Residues in Tobacco Plants" to the Tobacco Growers Association meeting in East Windsor, CT (February 24)(110 people). Dr. Brian Eitzer along with Dr. Jason White hosted a visit with Michael Rickenbach, Robert Lockwood, and Anuja Bharadwaj of the Chemistry Section of the State of CT - Department of Emergency Services & Public Protection to discuss high resolution liquid chromatography/mass spectrometry (March 26). Dr. Brian Eitzer was an instructor for the class entitled "LB511 FDA/FERN Chemistry Training for LC/MS" which was held at the Food and Drug Administration's Forensic Chemistry Center in Cincinnati, OH from June 21-26 (13 students) Dr. Brian Eitzer presented a poster on the use of Liquid Chromatography High Resolution Mass Spectrometry at 52nd Annual North American Chemical Residue Workshop in St. Petersburg Beach, FL (350 attendees at meeting) (July19 -July 23). Dr. Brian Eitzer presented a talk entitled "The Analysis of Pesticide Residues in Foods using Liquid Chromatography and High Resolution Mass Spectrometry" (25 attendees) at the Eastern Analytical Symposium in Somerset, NJ (November 16-18). What do you plan to do during the next reporting period to accomplish the goals?The primary goal during the next reporting period will be to complete the full validation of the multi-residue pesticide method which has been currently started. We plan to continue use of this instrumentation as our primary method of analyzing pesticide residues. Secondary goals are to continue to add contaminants to the list of compounds that can be screened for. A third goal is to evaluate the use of a new screening software program, ToxFinder which was recently received. Although similar to Tracefinder this program may simplify the initial screening process used in the identification of pesticides.

Impacts
What was accomplished under these goals? During the past year we have made the switch to HRMS on our older Thermo Exactive as the primary instrument used to quantify pesticide residues in produce and environmental samples. Produce samples are analyzed after a full QuEChERS based extraction and cleanup, vegetation and soils are typically analyzed after the initial salt out phase of the QuEChERS, and water samples are analyzed after a solid phase extraction. The pesticides are tentatively identified using Tracefinder EFS in the screening mode. In the quantitation mode of this software compounds are confirmed by examination of the integrated peaks for both quantitative and confirmation ions both of which must be present for a compound to be reported. The analysis of pesticide residues in produce is undergoing a full validation as part of our ISO accreditation. The initial analysis of this validation indicates that the full uncertainty of the method (extraction plus instrumental analysis) will be on the order of 30% for all samples throughout the entire calibration range. A second focus during this past year has been on initial testing of a newly received HRMS instrument, a Thermo Q-Exactive. This instrument incorporates a quadrupole ion filter before the orbitrap. This additional filter allows more modes of analysis than our current Exactive. The older instrument can only do all ion fragmentation, while the newer version can isolate single ions, or ion ranges to perform true MS-MS. We have been exploring these alternate modes of analysis in a project in which we are analyzing a set of 12 mycotoxins in corn, wheat and peanut butter.

Publications

Progress 10/01/13 to 09/30/14

Outputs
Target Audience: The primary target audience for this project are regulators and other scientists whom have an interest in the protection of the environment and the food supply. During this period, the target audience reached were principally other scientists present at the presentations listed under dissemination. Changes/Problems: A new high resolution mass spectrometer was recently received by the Connecticut Agricultural Experiment Station. This instrument is called a Thermo Q-Exactive and is an upgrade on the current orbitrap system used for this project. This newinstrument is both more sensitive and selective than the current instrument. The added selectivity comes from having a quadrupole installed in the area before the orbitrap. This quadrupole will allow a specific ion to be isolated so that in addition to the all ion MS/MS spectra produced by the current Exactive, a specific ion MS/MS spectrum can be obtained unequivocally linking a specific parent and product ion. The MS/MS spectra produced by this instrument should be similar to those produced on the current HRMS system, but this will be confirmed. A major goal for the next reporting period will be to learn the use of this instrument and the software associated with it so that we can develop test methods and compare results produced by this instrument with the previous instrument. What opportunities for training and professional development has the project provided? Dr. Brian Eitzer attended a seminar conducted by AB Sciex on new developments in forensic toxicology using LC/MS/MS at Yale Medical School How have the results been disseminated to communities of interest? Jan. 10-11 Dr. Eitzer gave a talk entitled “Analysis of Pesticides in Planter Exhaust Dust and Dosimeters Surrounding Fields During Planting” at the American Bee Research Conference in San Antonio, TX Jan 10-11 (50 people) June 5 Dr. Eitzer gave a webinar on the analysis of aflatoxin M1 in milk using a Thermo Velos to the FERN cCAP Mycotoxin Working Group on (20 attendees) June 20 Dr. Eitzer gave a presentation on the detection of aflatoxins in milk at the monthly Toxi-Rounds at the Department of Health Laboratory in Rocky Hill (15 attendees) June 30-July 3 Dr. Eitzer presented a poster on the analysis of pesticides in produce and teas at the European Pesticide Residue Workshop in Dublin Ireland (500 attendees) Sept. 7-10 Dr. Eitzer attended and presented a poster on the analysis of aflatoxins in milk at the AOAC International Meeting in Boca Raton FL (900 attendees) Oct. 23 Dr. Eitzer gave a webinar on “Analysis of Pesticide Residues in Tea Leaves” to the Northeast FERN group on (15 people) What do you plan to do during the next reporting period to accomplish the goals? We monitor the pesticides and pesticide metabolites available to us through the EPA repository. As the availability of pesticides that we have not tested is announced, we obtain these materials and test whether or not the can be determined by liquid chromatography/mass spectrometry. We will continue to do this work. A new effort during the next reporting period will be to learn the use of new second high resolution mass spectrometric instrumentat that we have recently obtained. This information is provided in the changes section below.

Impacts
What was accomplished under these goals? During the past year we have continued to examine whether specific pesticides and pesticide metabolites can be observed with a liquid chromatography high resolution mass spectrometric system (HRMS). Using both positive and negative electrospray modes, there are currently over 750 different compounds included in the targeted lists we use. Over 150 samples of groundwaters, soils and vegetation from various investigations were screened for pesticides using high resolution mass spectrometry after either a solid phase extraction of the water samples or a QuEChERS-based acetonitrile extraction of the solid matrix. We also continue to screen for pesticides in bee related matrixes using this technique. We have been exploring the use of the HRMS as the primary quantitation instrument for the LC/MS pesticides reported in a market basket survey program in the State of Connecticut. One new effort in the past year has been the investigation of a software package called Tracefinder EFS. This package has some advantages in the processing of the data that allows easier access to quality assurance/quality control information within the raw data. The added QA/QC capabilities will be important as we plan on seeking ISO accreditation for this work

Publications

Progress 01/01/13 to 09/30/13

Outputs
Target Audience: The principal target audiences for this work are regulators and other scientists whom have an interest in the protection of the food supply. They have a need to know of improved methods for the detection of contaminants as well as the actual reports of contaminants in foods. The public also needs to be reassured in the safety of the food supply. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Nothing Reported How have the results been disseminated to communities of interest? The project director presented a talk entitled “The Connecticut Market Basket Survey for Pesticide Residues in Produce” at the 50th annual North American Chemical Residue Workshop in St. Petersburg Beach Fl, July 21-24. (150 attendees) The project director resented a talk entitled “Testing and Validation of T022 using High Resolution Mass Spectrometry” and at the FERN cCAP annual meeting in St. Paul, MN Sept. 17-19. (50 attendees) The project director resented a talk entitled “The determination of Pesticides and other Contaminants in Foods” to the Yale Occupational and Environmental Medicine Group” on Dec. 10 (15 attendees) What do you plan to do during the next reporting period to accomplish the goals? During the next reporting period we plan to continue adding compounds to the screening list. A second item we plan on working on during the next reporting period is an improvement in the process of reviewing and reporting the data produced. One drawback of the current system is the extensive amount of time spent needed to review data for over 700 compounds in each sample analyzed. The current screen uses ToxID software to help with this review, but, still requires an extensive manual data review. We recently acquired a new software program (TraceFinder) that can be used in the analysis of high resolution mass spectrometry data. We plan on evaluating this software to see if it will allow us to reduce the amount of time used for data review.

Impacts
What was accomplished under these goals? This report covers the time period from January 1, 2013 - Dec 31, 2013. During this time period we have continued to add compounds to the list of chemicals in the high resolution mass spectrometry pesticidescreen. This list now numbers over 700 compounds, though, it should be noted that the list just incorporates any compound which was detected by the mass spectrometer in the initial high concentration screening. Thus, there are compounds on the list which respond poorly or do not extract by either the QuEChERS or solid phase extraction techniques typically used. This year the high resolution instrumentation was used to analyze 90 surface waters for organic contaminants. The water samples were prepared by solid phase extraction and then injected on the instrument in both positive and negative ion modes. It was found that the instrumentation was able to produce good quantitative data in both modes with detection limits for many compounds in the 0.01 ppb range. We are also comparing the data produced by the high resolution instrument with data produced by our unit resolution MS/MS system in the analysis of exhaust dust from corn planters, which may be toxic to bees,and dosimeters placed around the field during sampling. During these analyses the two instruments are producing similar data, though there are more compounds that could be potentially seen withthe high resolution instrument.

Publications

• Type: Journal Articles Status: Awaiting Publication Year Published: 2014 Citation: Eitzer, B.D.; Hammack, W.; Filigenzi, M. 2014. Interlaboratory Comparison of a general method to screen foods for pesticides using quechers extraction with high performance liquid chromatography and high resolution mass spectrometry. J Agr Food Chem. DOI 10.1021/jf405128y

Progress 01/01/12 to 12/31/12

Outputs
OUTPUTS: This new project is designed to improve the pesticide residue (and other organic contaminants) monitoring procedures of the Connecticut Agricultural Experiment Station by both increasing the number of individual residues that can be monitored as well as improving detection limits for other residues. Several mixed standards have been obtained, which contain hundreds of different pesticide residues. In addition, a mixture of 15 mycotoxins has also been obtained. These mixtures have been screened using high resolution liquid chromatography/high resolution mass spectrometry. We are evaluating operating conditions (resolution, MS/MS conditions, ionization mode etc.) that provide for the best identification and quantitation for a broad range of pesticides. We are currently analyzing all our pesticide residues in produce samples using both our older unit resolution MS/MS system and the new high resolution MS/MS system to compare the capabilities of the two systems. Events and Dissemination: July 16, "The Use of High Performance Liquid Chromatography/High Resolution Mass Spectrometry in Screening Produce for Pesticide Residues" at the Florida Pesticide Residue Workshop, St. Petersburg Beach FL. Sept 12, "QuEChERS Exactive Pesticide Screening Protocol" at the Food Emergency Response Networks cCAP meeting, Cincinnati, OH. Oct. 9 and Oct. 11 "The Use of High Performance Liquid Chromatography/High Resolution Mass Spectrometry in Screening Produce for Pesticide Residues" at the Thermo-Fisher Scientific LC/MS User Meetings in Somerset, NJ, and Cambridge, MA. PARTICIPANTS: Dr. Brian Eitzer is the principal scientist working on this project. Some of the samples to be analyzed are prepared by Dr. Walter Krol and Ms. Teri Arsenault. TARGET AUDIENCES: The primary target audiences for this Hatch project are the agencies, such as the Connecticut Department of Consumer Protection, the Connecticut Department of Energy and Environmental Protection and the U.S. Food and Drug Administration that use the pesticide residue data generated by the Connecticut Agricultural Experiment Station. Secondary target audiences include the public whom are interested in the safety of the food supply. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.

Impacts
The goal of this project is the enhancement of the pesticide residue monitoring program of the Connecticut Agricultural Experiment Station. During the past year, seven pesticide residues were found for the first time in our ongoing survey of pesticide residues in Connecticut produce. These residues were only identified because of the high resolution mass spectrometry procedures being introduced as part of this project. These residues are cadusaphos, carbosulfan, fenuron, kresoxim-methyl, spinosad, and spiridoclofen. Furthermore, three samples, which contained some of these residues, were reported to the Connecticut Department of Consumer Protection (DCP)as being violative; these were a turnip sample from Massachusetts which had 0.002 ppm fenuron, a basil sample from Columbia with 0.017 ppm cadusafos and a pepper sample from China with 0.089 ppm carbosulfan. In all three cases there was no tolerance for these residues on that commodity. As none of these commodities were from Connecticut the DCP passed the information on o the Food and Drug Administration for further action.

Publications

• No publications reported this period