ACQUISITION OF UHPLC-MS/MS INSTRUMENTATION TO STRENGTHEN INTERDISCIPLINARY RESEARCH ON BIOMOLECULES AND EMERGING CONTAMINANTS IN AGROECOSYSTEMS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: TERMINATED
• Funding Source: OTHER GRANTS
• Reporting Frequency: Annual
• Accession No.: 1026942
• Grant No.: 2021-70410-35298
• Project No.: NH.W-2021-05013
• Proposal No.: 2021-05013
• Program Code: EGP
• Project Start Date: Jul 1, 2021
• Project End Date: Jun 30, 2023
• Grant Year: 2021

Project Director
Grandy, A. S.

Recipient Organization
UNIVERSITY OF NEW HAMPSHIRE
51 COLLEGE RD SERVICE BLDG 107
DURHAM,NH 03824

Performing Department
NREN

Non Technical Summary
Our proposal requests an ultra-high-performance high resolution liquid chromatography-tandem mass spectrometry instrument (UHPLC-MS/MS), bringing together multiple University of New Hampshire (UNH) research programs in a collaborative effort to understand the dynamics of natural soil organic matter (SOM) and anthropogenic organic contaminants in agroecosystems. SOM is inextricably tied to soil and ecosystem health but rapidly evolving concepts of SOM formation and the pressing need to track anthropogenic 'compounds ofemerging concern' (CECs) demand state-of-the-art chemical analysis. The proposed UHPLC-MS/MS instrumentation will alleviate UNH's current inability to quantify specific organic biomolecules and CECs, and will allow us to develop integrated applications tocharacterize their interactions, track them into plant-soil systems and water resources, gauge their availability to microbial communities, and understand the forces governing their persistence in soils. Our proposed research addresses the USDA Science Blueprint's roadmap for 2020 to 2025 Program Themes 1) Agricultural Intensification and 2) Agricultural Climate Adaptation by enablingnew SOM insights that become the foundation for high-yielding and resilient agroecosystems. We also contribute to Program Themes 3) Food and Nutrition Translation by enabling us to determine the extent to which CECs may threaten food safety, and 5) Agriculture Science Policy Leadership by generating data that regulatory agencies can use to shape applied research and new policies. Our discoveries about the source and cycling of natural SOM and anthropogenic CECs, their underlying microbial processes, and their ecosystem consequences will have real-world significance for industry and regional agricultural stakeholders. ?We will purchase a high-performance high resolution liquid chromatography-tandem mass spectrometry instrument (UHPLC-MS/MS), bringing together multiple University of New Hampshire (UNH) research programs in a collaborative effort to understand the dynamics of natural soil organic matter (SOM) and anthropogenic organic contaminants in agroecosystems. SOM is inextricably tied to soil and ecosystem health but rapidly evolving concepts of SOM formation and the pressing need to track anthropogenic 'compounds of emerging concern' (CECs) demand state-of-the-art chemical analysis. The proposed UHPLC-MS/MS instrumentation will alleviate UNH's current inability to quantify specific organic biomolecules and CECs, and will allow us to develop integrated applications to characterize their interactions, track them into plant-soil systems and water resources, gauge their availability to microbial communities, and understand the forces governing their persistence in soils. Our proposed research addresses the USDA Science Blueprint's roadmap for 2020 to 2025 Program Themes 1) Agricultural Intensification and 2) Agricultural Climate Adaptation by enabling new SOM insights that become the foundation for high-yielding and resilient agroecosystems. We also contribute to Program Themes 3) Food and Nutrition Translation by enabling us to determine the extent to which CECs may threaten food safety, and 5) Agriculture Science Policy Leadership by generating data that regulatory agencies can use to shape applied research and new policies. Our discoveries about the source and cycling of natural SOM and anthropogenic CECs, their underlying microbial processes, and their ecosystem consequences will have real-world significance for industry and regional agricultural stakeholders.

Animal Health Component

10%

Research Effort Categories

Basic

60%

Applied

20%

Developmental

20%

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
101	0110	1000	50%
133	0110	2000	50%

Knowledge Area
133 - Pollution Prevention and Mitigation; 101 - Appraisal of Soil Resources;

Subject Of Investigation
0110 - Soil;

Field Of Science
1000 - Biochemistry and biophysics; 2000 - Chemistry;

Keywords

mass spectrometry

pfas

contaminants

lipids

soil organic matter

Goals / Objectives
We will purchase and install an ultra-high-performance high resolution liquid chromatography-tandem mass spectrometry instrument (UHPLC-MS/MS), bringing together multiple University of New Hampshire (UNH) research programs in a collaborative effort to understand the dynamics of natural soil organic matter (SOM) and anthropogenic organic contaminants in agroecosystems. SOM is inextricably tied to soil and ecosystem health but rapidly evolving concepts of SOM formation and the pressing need to track anthropogenic 'compounds of emerging concern' (CECs) demand state-of-the-art chemical analysis. The proposed UHPLC-MS/MS instrumentation will alleviate UNH's current inability to quantify specific organic biomolecules and CECs, and will allow us to develop integrated applications to characterize their interactions, track them into plant-soil systems and water resources, gauge their availability to microbial communities, and understand the forces governing their persistence in soils.

Project Methods
The proposed instrument will be housed in our University Instrumentation Center (UIC) Mass Spectrometry Core Facility, an FDA registered facility that already has document control, training procedures, and calibration programs in place. We will fund ongoing operating costs--encompassing consumables, service contracts, labor, maintenance, repair, and user scheduling--through per-sample fees carefully considered to cover expenses while maximizing accessibility to our community of users. This model is in place and proven to fund the UIC MS Core without overhead to users.UNH has, and continues to demonstrate, a strong commitment to maintain and improve its research instrumentation. The UIC MS Core Facility will assist with instrument maintenance, repair, and scheduling. Annual expected operating costs for the instrumentation include mass spectrometry specific gases and LC consumables (columns, spray tips, solvents, standards, etc.). At this time, the annual operating costs, based on current pricing and excluding labor cost, will be ~ $90,000 (with service contract). It is anticipated that instrument usage will approach 85-90%, and we anticipate charging ~$150 per sample for UNH users to cover operating costs. For non-UNH users, the rate will be twice that of UNH user fees, in accordance with current UIC operating practices. The rate per sample will be re-evaluated periodically and adjusted to keep it at a minimum, in order to ensure maximum accessibility for researchers while recovering operating expenses. Since 2015, the UIC has helped generate and oversee invoices for MS analyses, without overhead to users. A dedicated account has been created for the MS Core Facility service. Without purchasing an MS service contract and paying service technician labor, the account has maintained proper balances for repair needs. Revenues were generated mainly from the PI and major users' samples.Throughout the project, data will be stored and archived by the Research Computing Center (RCC), a UNH Core Facility housed on the UNH campus. PI Grandy will work with the RCC Director, Patrick Messer, to ensure proper and efficient data management and long-term preservation. RCC support will include data storage on secure servers in the RCC data center, system and network administration and security, data backups, archiving, and remote data replication to the RCC storage system at the UNH IT Data Center. Team members on the project will be able to access the data via remote access set up by the RCC. Researchers can remotely access their data and results and securely download using standard open source technologies. Computational scientists at UNH RCC will maintain the advanced MS data processing and analysis software tools, as well as help train users in bioinformatics.

Progress 07/01/22 to 06/30/23

Outputs
Target Audience:The target audience for the activities in this grant related to the new instrument installation includes graduate students in analytical chemistry seeking to acquire cutting-edge skills and faculty members looking to broaden their research capabilities in lipidomics, PFAS analysis, and metabolomics. The new capabilities are also beginning to lead to collaborations between stakeholders interested in fostering public-private partnerships and university personnel. This diverse group is already benefitting from the enhanced research opportunities and educational advancements that the new instrument has created, leading to new collaborations and research in agricultural and environmental sciences. Changes/Problems:There were fairly typical universitychallenges encountered in ordering, locating, and setting up the new analytical instrument, as well as in establishing the necessary protocols. Given this, we were somewhat delayed in other areas, including generating robust external collaborations. We now have momentum in this area, as well as with expanding internal use of the instrument to include teaching and a range of researchers. What opportunities for training and professional development has the project provided?Our accomplishments indicate multiple users trained on the instrument, in addition to the following formal training. 3-Day Application and TraceFinder software training provided by Thermo. (UIC, Mouser and Grandy Groups in attendance) (03/2023) Thermo e-learning courses (UIC, on demand) LC-MS users are trained on LC-MS operation by UIC personnel. Training is based on the operator's level of experience with LC-MS and their project requirements. How have the results been disseminated to communities of interest?We established a website for external contacts and collaboration (Mass Spectrometer - Thermo Scientific Exploris 120 University Instrumentation Center (unh.edu)), containing the following, The Exploris 120 is a high-resolution heated electrospray mass spectrometer that operatesover a mass range of m/z 40 - 3,000 and is coupled witha Vanquish UHPLC system to perform LC-MS analysis. Using an orbitrap analyzer, the instrument is capable of up to 120,000 FWHM mass resolving power (at m/z 200), a < 1 ppm mass error, and offers sensitivities on the order of femtograms.The orbitrap detector also allows for a largedynamic range of up to >5,000, with the instrument electronics capable of high scan rates and polarity switching. In addition, the instrument is capable of different MS/MS experiments in order to elucidate molecular structures of small molecules. We have had multiple new user inquiries in the past month based on the website and word of mouth, including internal UNH users and potential external public and private users. What do you plan to do during the next reporting period to accomplish the goals?Given the challenges encountered in ordering, locating, and setting up the new analytical instrument, as well as in establishing the necessary protocols, efforts have now pivoted towards expanding internal and external collaborations. Current and future collaborations span a diverse range of areas, including metabolomics, soil lipids, and PFAS analysis. The audience for these collaborations will continue to include graduate students in analytical chemistry, faculty researchers, and partners interested in public-private collaborations. We will continue to leverage this state-of-the-art instrument to enhance research capabilities, educational opportunities, and interdisciplinary partnerships in critical agricultural and environmental fields.

Impacts
What was accomplished under these goals? After some delays, we purchased the ultra-high-performance high-resolution liquid chromatography-tandem mass spectrometry instrument (UHPLC-MS/MS). The shipping was delayed, but the instrument arrived and became fully operational spring-summer 2023. A newly hired pHD-level Research Scientist, a leader of the University Instrumentation Center Mass Spec Core, Philip Place, was crucial to setting up the instrument and establishing new protocols. Since its installation, there have been methods development and new training and collaborations involving internal and external partners and classroom teaching. Activities include the following. Paula Mouser and Stuart Grandy, Optimized methods for soil lipidomics based on collaboration with EMSL, run various soils and identified dozens of lipids. Initiated methods development for PFAS. Liz Harvey - UNH Biological Sciences (1 PostDoc) - Ocean Metabolomics, Started 04/2023, Preliminary project was successful by 08/2023, Started on full project Fall 2023 with samples ready by 02/2024 Anyin Li - UNH Chemistry (3 Graduate Students) - Femtoflow and Native ESI Development, Started 01/2024 Glen Miller- UNH Chemistry (1 Graduate Student) - Molecular Structure Verification, Started 07/2023 External Client - Chemical Purity Verification, Started 12/2023 Chemistry Course (Chem 763) - Teaching Lab Module developed for Spring semester 2024. Started Methods Transfer 12/2023, Use in teaching Labs planned for 03/2024 We developed anadvanced analytical protocol in conjunction with EMSL for soil lipid extraction based on a Modified Folch protocoland subsequent analysis on the ms, including thedevelopment of a lipids library.

Publications

Progress 07/01/21 to 06/30/23

Outputs
Target Audience:The target audience for the activities in this grant related to the new instrument installation includes graduate students in analytical chemistry seeking to acquire cutting-edge skills and faculty members looking to broaden their research capabilities in lipidomics, PFAS analysis, and metabolomics. The new capabilities are also beginning to lead to collaborations between stakeholders interested in fostering public-private partnerships and university personnel. This diverse group is already benefitting from the enhanced research opportunities and educational advancements that the new instrument has created, leading to new collaborations and research in agricultural and environmental sciences. Changes/Problems:There were fairly typical university challenges encountered in ordering, locating, and setting up the new analytical instrument, as well as in establishing the necessary protocols. Given this, we were somewhat delayed in other areas, including generating robust external collaborations. We now have momentum in this area, as well as with expanding internal use of the instrument to include teaching and a range of researchers. What opportunities for training and professional development has the project provided?Our accomplishments indicate multiple users trained on the instrument, in addition to the following formal training. 3-Day Application and TraceFinder software training provided by Thermo. (UIC, Mouser and Grandy Groups in attendance) (03/2023) Thermo e-learning courses (UIC, on demand) LC-MS users are trained on LC-MS operation by UIC personnel. Training is based on the operator's level of experience with LC-MS and their project requirements. How have the results been disseminated to communities of interest?We established a website for external contacts and collaboration (Mass Spectrometer - Thermo Scientific Exploris 120 University Instrumentation Center (unh.edu)), containing the following, The Exploris 120 is a high-resolution heated electrospray mass spectrometer that operates over a mass range of m/z 40 - 3,000 and is coupled with a Vanquish UHPLC system to perform LC-MS analysis. Using an orbitrap analyzer, the instrument is capable of up to 120,000 FWHM mass resolving power (at m/z 200), a < 1 ppm mass error, and offers sensitivities on the order of femtograms. The orbitrap detector also allows for a large dynamic range of up to >5,000, with the instrument electronics capable of high scan rates and polarity switching. In addition, the instrument is capable of different MS/MS experiments in order to elucidate molecular structures of small molecules. We have had multiple new user inquiries in the past month based on the website and word of mouth, including internal UNHusers and potential external public and private users. What do you plan to do during the next reporting period to accomplish the goals?Given the challenges encountered in ordering, locating, and setting up the new analytical instrument, as well as in establishing the necessary protocols, efforts have now pivoted towards expanding internal and external collaborations. Current and future collaborations span a diverse range of areas, including metabolomics, soil lipids, and PFAS analysis. The audience for these collaborations will continue to include graduate students in analytical chemistry, faculty researchers, and partners interested in public-private collaborations. We will continue to leverage this state-of-the-art instrument to enhance research capabilities, educational opportunities, and interdisciplinary partnerships in critical agricultural and environmental fields.

Impacts
What was accomplished under these goals? After some delays, we purchased the ultra-high-performance high-resolution liquid chromatography-tandem mass spectrometry instrument (UHPLC-MS/MS). The shipping was delayed, but the instrument arrived and became fully operational spring-summer 2023. A newly hired pHD-level Research Scientist, a leader of the University Instrumentation Center Mass Spec Core, Philip Place, was crucial to setting up the instrument and establishing new protocols. Since its installation, there have been methods development and new training and collaborations involving internal and external partners and classroom teaching. Activities include the following. Paula Mouser and Stuart Grandy, Optimized methods for soil lipidomics based on collaboration with EMSL, run various soils and identified dozens of lipids. Initiated methods development for PFAS. Liz Harvey - UNH Biological Sciences (1 PostDoc) - Ocean Metabolomics, Started 04/2023, Preliminary project was successful by 08/2023, Started on full project Fall 2023 with samples ready by 02/2024 Anyin Li - UNH Chemistry (3 Graduate Students) - Femtoflow and Native ESI Development, Started 01/2024 Glen Miller- UNH Chemistry (1 Graduate Student) - Molecular Structure Verification, Started 07/2023 External Client - Chemical Purity Verification, Started 12/2023 Chemistry Course (Chem 763) - Teaching Lab Module developed for Spring semester 2024. Started Methods Transfer 12/2023, Use in teaching Labs planned for 03/2024 We developed an advanced analytical protocol in conjunction with EMSL for soil lipid extraction based on a Modified Folch protocol and subsequent analysis on the ms, including the development of a lipids library.

Publications

Progress 07/01/21 to 06/30/22

Outputs
Target Audience:At this point, the target audience are scientists and professional staff ordering the instrument, setting it up, and establishing protocols. Changes/Problems:We have had some delays. We had to identify a new room location that met all instrument specifications. We had to hire the scientist. There were multiple delays getting the instrument specification and pricing finalized with Thermo. The most substantial delay was with shipping due to pandemic-related back orders. But these have been overcome. What opportunities for training and professional development has the project provided?Once the instrument arrives we have paid Thermo Fisher for training. After this protocols will be established in 2023 and new participants engaged. How have the results been disseminated to communities of interest? Nothing Reported What do you plan to do during the next reporting period to accomplish the goals?In the following reporting period we will: install the instrument, receive instrument training, begin to establish SOM and PFAS protocols, and engage students in potential projects.

Impacts
What was accomplished under these goals? After some delays we purchased the ultra-high-performance high resolution liquid chromatography-tandem mass spectrometry instrument (UHPLC-MS/MS). The shipping was delayed but the instrument has arrived. The instrument will be arrived Fall, 2022. After that, protocols will be established.

Publications