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Independent Review of Non-Destructive Assay for the K-25/K-27 D&D Project: November 30-December 3, 2004, Oak Ridge, TN

The DOE with concurrence from the D&D contractor has chartered an assessment of the methods and procedures used to obtain the original NDA results of holdup deposits at the K-25 and K-27 uranium gaseous diffusion facilities in Oak Ridge. The assessment has been performed by an external review team with the participation of the D&D contractor, DOE, and DOE/OR. This document is the final report for the assessment. The DOE provided the charter for this review. The review team spent a week in Oak Ridge attending meetings and participating in discussions with expert contractor staff. Substantial additional input for the review came from numerous written materials, unpublished and published. This final report of observations, findings, and recommendations in areas defined by the charter is based on information from the meetings, discussions, and documents. The assessment of the Review Team is that the DOE/OR and BJC approach in using historical NDA data for the D&D of K-25 and K-27 is appropriate and generally acceptable. Resolving issues, addressing findings, and implementing the recommendations documented in this report will reconcile specific technical concerns.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Development and implementation of high-throughput proteomic and metabolomics assays by using advanced chromatographic and mass spectrometric systems (CRADA Final Report)

The mission of this CRADA with Agilent was to couple powerful MS platforms (QQQ, IM-QTOFMS) with Agilent’s novel Ultra-High-Performance Liquid Chromatography (UHPLC) fast metabolomic workflows and perform ABF Machine Learning (ML) to generated datasets. Agilent transferred UHPLC methods to PNNL and LBNL and methods were implemented and demonstrated in both labs, achieving total acquisition times of < 10 min. Metabolites analyzed using Agilent’s shared methods included metabolites from central carbon metabolism, common across hosts, and metabolites unique to engineered strains. Standards were acquired in an UHPLC-Drift Tube Ion Mobility Mass Spectrometer (DTIMS) system for the first time within the context of ABF and methods were optimized based on Agilent’s protocols. Samples from ABF hosts Pseudomonas putida, Aspergillus pseudoterreus, Aspergillus niger and Rhodosporidium toruloides were analyzed using the UHPLC-DTIMS platform for a total of 276 runs. A data analysis workflow compatible with the Experimental Data Depot (EDD) and completely shareable was developed for the acquired UHPLC-DTIMS data. Samples were analyzed using a Data Independent Acquisition Approach (DIA), which for most of the standards provided more transitions therefore increasing detection confidence. Using the data acquired by PNNL, LBNL, and Agilent’s specifications from previous ML projects, SNL applied an ensemble ML strategy to pick the best performing model for automated LC-method selection. Finally, with the contribution of the participant labs and Agilent, SNL developed an Automated Method Selection (AMS) software tool to predict the best liquid chromatography method for analysis of any new molecules of interest. Samples with novel pathways and new metabolite targets of interest are generated at a high pace in the ABF. Overall, the project advanced rapid metabolomics by combining liquid chromatography, ion mobility spectrometry, and data-independent mass spectrometry with machine learning. This multidimensional approach uses retention time, collision cross-section, precursor mass, and fragment-ion information to distinguish chemically similar metabolites that can be difficult to resolve using conventional liquid- or gas-chromatography methods. The resulting workflow also provided automated metabolite-identification error estimates, addressing a recognized need for statistical confidence measures in metabolomics.

Petzold, Christopher [Lawrence Berkeley National L↗