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DOE OSTI2023

Alkaline-SDS cell lysis of microbes with acetone protein precipitation for proteomic sample preparation in 96-well plate format

Plate-based proteomic sample preparation offers a solution to the large sample throughput demands in the biotechnology field where hundreds or thousands of engineered microbes are constructed for testing is routine. Meanwhile, sample preparation methods that work efficiently on broader microbial groups are desirable for new applications of proteomics in other fields, such as microbial communities. Here, we detail a step-by-step protocol that consists of cell lysis in an alkaline chemical buffer (NaOH/SDS) followed by protein precipitation with high-ionic strength acetone in 96-well format. The protocol works for a broad range of microbes ( e . g ., Gram-negative bacteria, Gram-positive bacteria, non-filamentous fungi) and the resulting proteins are ready for tryptic digestion for bottom-up quantitative proteomic analysis without the need for desalting column cleanup. The yield of protein using this protocol increases linearly with respect to the amount of starting biomass from 0.5–2.0 OD*mL of cells. By using a bench-top automated liquid dispenser, a cost-effective and environmentally-friendly option to eliminating pipette tips and reducing reagent waste, the protocol takes approximately 30 minutes to extract protein from 96 samples. Tests on mock mixtures showed expected results that the biomass composition structure is in close agreement with the experimental design. Lastly, we applied the protocol for the composition analysis of a synthetic community of environmental isolates grown on two different media. This protocol has been developed to facilitate rapid, low-variance sample preparation of hundreds of samples and allow flexibility for future protocol development.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2014

Raw neutron scattering data for strain measurement of hydraulically loaded granite and marble samples in triaxial stress state

This entry contains raw data files from experiments performed on the Vulcan beamline at the Spallation Neutron Source at Oak Ridge National Laboratory using a pressure cell. Cylindrical granite and marble samples were subjected to confining pressures of either 0 psi or approximately 2500 psi and internal pressures of either 0 psi, 1500 psi or 2500 psi through a blind axial hole at the center of one end of the sample. The sample diameters were 1.5" and the sample lengths were 6". The blind hole was 0.25" in diameter and 3" deep. One set of experiments measured strains at points located circumferentially around the center of the sample with identical radii to determine if there was strain variability (this would not be expected for a homogeneous material based on the symmetry of loading). Another set of experiments measured load variation across the radius of the sample at a fixed axial and circumferential location. Raw neutron diffraction intensity files and experimental parameter descriptions are included.

15 GEOTHERMAL ENERGY↗

DOE OSTI2021

G-LiHT Campaign Leaf Sample details & photos, March 2017: Puerto Rico

This data package includes details of leaves sampled for leaf spectra and chemistry from 5 sites in Puerto Rico, in March of 2017. Sunlit canopy and shaded leaves of 66 species were collected. Data for each sample includes species, leaf age, type of analysis (spectroscopy, gas exchange, chemistry), sample number and sample photographs. The data package includes a spreadsheet with sample information and a zip file of photographs (1.6 GB). This data was collected as part of the 2017 BNL–G-LiHT leaf spectra campaign. See related datasets for leaf spectral reflectance and transmittance, leaf mass area (LMA), and leaf chemistry. Note that leaf sample details are also included in related datasets.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2012

A General Investigation of Optimized Atmospheric Sample Duration

ABSTRACT The International Monitoring System (IMS) consists of up to 80 aerosol and xenon monitoring systems spaced around the world that have collection systems sensitive enough to detect nuclear releases from underground nuclear tests at great distances (CTBT 1996; CTBTO 2011). Although a few of the IMS radionuclide stations are closer together than 1,000 km (such as the stations in Kuwait and Iran), many of them are 2,000 km or more apart. In the absence of a scientific basis for optimizing the duration of atmospheric sampling, historically scientists used a integration times from 24 hours to 14 days for radionuclides (Thomas et al. 1977). This was entirely adequate in the past because the sources of signals were far away and large, meaning that they were smeared over many days by the time they had travelled 10,000 km. The Fukushima event pointed out the unacceptable delay time (72 hours) between the start of sample acquisition and final data being shipped. A scientific basis for selecting a sample duration time is needed. This report considers plume migration of a nondecaying tracer using archived atmospheric data for 2011 in the HYSPLIT (Draxler and Hess 1998; HYSPLIT 2011) transport model. We present two related results: the temporal duration of the majority of the plume as a function of distance and the behavior of the maximum plume concentration as a function of sample collection duration and distance. The modeled plume behavior can then be combined with external information about sampler design to optimize sample durations in a sampling network.

CTBT↗

DOE OSTI2013

Analytical Results for MOX Colemanite Concrete Samples Received on January 15, 2013

The Mixed Oxide Fuel Fabrication Facility (MFFF) will use colemanite bearing concrete neutron absorber panels credited with attenuating neutron flux in the criticality design analyses and shielding operators from radiation. The Savannah River National Laboratory (SRNL) is tasked with measuring the total density, partial hydrogen density, and partial boron density of the colemanite concrete. SRNL received twelve samples of colemanite concrete for analysis on January 15, 2013. The average total density of each of the samples measured by the ASTM method C 642, the average partial hydrogen density was measured using method ASTM E 1311, and the average partial boron density of each sample was measured according to ASTM C 1301. The lower limits and measured values for the total density, hydrogen partial density, and boron partial density are presented. For all the samples tested, the total density and the hydrogen partial density met or exceeded the specified limit. All of the samples met or exceeded the boron partial density lower bound with the exception of samples G3-M11-2000-H, G3-M11-3000-M, and G5-M1-3000-H which are below the limit of 1.65E-01 g/cm 3 .

36 MATERIALS SCIENCE↗

DOE OSTI2013

Results Of Routine Strip Effluent Hold Tank, Decontaminated Salt Solution Hold Tank, Caustic Wash Tank And Caustic Storage Tank Samples From Modular Caustic-Side Solvent Extraction Unit During Macrobatch 6 Operations

Strip Effluent Hold Tank (SEHT), Decontaminated Salt Solution Hold Tank (DSSHT), Caustic Wash Tank (CWT) and Caustic Storage Tank (CST) samples from several of the “microbatches” of Integrated Salt Disposition Project (ISDP) Salt Batch (“Macrobatch”) 6 have been analyzed for 238 Pu, 90 Sr, 137 Cs, and by Inductively Coupled Plasma Emission Spectroscopy (ICPES). The results from the current microbatch samples are similar to those from comparable samples in Macrobatch 5. From a bulk chemical point of view, the ICPES results do not vary considerably between this and the previous macrobatch. The titanium results in the DSSHT samples continue to indicate the presence of Ti, when the feed material does not have detectable levels. This most likely indicates that leaching of Ti from MST in ARP continues to occur. Both the CST and CWT samples indicate that the target Free OH value of 0.03 has been surpassed. While at this time there is no indication that this has caused an operational problem, the CST should be adjusted into specification. The 137 Cs results from the SRNL as well as F/H lab data indicate a potential decline in cesium decontamination factor. Further samples will be carefully monitored to investigate this.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2013

A margin based approach to determining sample sizes via tolerance bounds.

This paper proposes a tolerance bound approach for determining sample sizes. With this new methodology we begin to think of sample size in the context of uncertainty exceeding margin. As the sample size decreases the uncertainty in the estimate of margin increases. This can be problematic when the margin is small and only a few units are available for testing. In this case there may be a true underlying positive margin to requirements but the uncertainty may be too large to conclude we have sufficient margin to those requirements with a high level of statistical confidence. Therefore, we provide a methodology for choosing a sample size large enough such that an estimated QMU uncertainty based on the tolerance bound approach will be smaller than the estimated margin (assuming there is positive margin). This ensures that the estimated tolerance bound will be within performance requirements and the tolerance ratio will be greater than one, supporting a conclusion that we have sufficient margin to the performance requirements. In addition, this paper explores the relationship between margin, uncertainty, and sample size and provides an approach and recommendations for quantifying risk when sample sizes are limited.

Newcomer, Justin T.↗

DOE OSTI2013

COMPARISON OF RESULTS FOR QUARTER 4 SURFACE WATER SPLIT SAMPLES COLLECTED AT THE NUCLEAR FUELS SERVICES SITE, ERWIN, TN

Oak Ridge Associated Universities (ORAU), under the Oak Ridge Institute for Science and Education (ORISE) contract, collected split surface water samples with Nuclear Fuel Services (NFS) representatives on June 12, 2013. Representatives from the U.S. Nuclear Regulatory Commission (NRC) and the Tennessee Department of Environment and Conservation were also in attendance. Samples were collected at four surface water stations, as required in the approved Request for Technical Assistance number 11-018. These stations included Nolichucky River upstream (NRU), Nolichucky River downstream (NRD), Martin Creek upstream (MCU), and Martin Creek downstream (MCD). Both ORAU and NFS performed gross alpha and gross beta analyses, and Table 1 presents the comparison of results using the duplicate error ratio (DER), also known as the normalized absolute difference. A DER ≤ 3 indicates at a 99% confidence interval that split sample results do not differ significantly when compared to their respective one standard deviation (sigma) uncertainty (ANSI N42.22). The NFS split sample report specifies 95% confidence level of reported uncertainties (NFS 2013). Therefore, standard two sigma reporting values were divided by 1.96. In conclusion, most DER values were less than 3 and results are consistent with low (e.g., background) concentrations. The gross beta result for sample 5198W0014 was the exception. The ORAU gross beta result of 6.30 &plusmn 0.65 pCi/L from location NRD is well above NFS's non-detected result of 1.56 &plusmn 0.59 pCi/L. NFS's data package includes no detected result for any radionuclide at location NRD. At NRC's request, ORAU performed gamma spectroscopic analysis of sample 5198W0014 to identify analytes contributing to the relatively elevated gross beta results. This analysis identified detected amounts of naturally-occurring constituents, most notably Ac-228 from the thorium decay series, and does not suggest the presence of site-related contamination.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2012

Gasbuggy, New Mexico, Natural Gas and Produced Water Sampling Results for 2012

The U.S. Department of Energy (DOE) Office of Legacy Management conducted annual natural gas sampling for the Gasbuggy, New Mexico, Site on June 20 and 21, 2012. This long-term monitoring of natural gas includes samples of produced water from gas production wells that are located near the site. Water samples from gas production wells were analyzed for gamma-emitting radionuclides, gross alpha, gross beta, and tritium. Natural gas samples were analyzed for tritium and carbon-14. ALS Laboratory Group in Fort Collins, Colorado, analyzed water samples. Isotech Laboratories in Champaign, Illinois, analyzed natural gas samples.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2014

Results Of Routine Strip Effluent Hold Tank, Decontaminated Salt Solution Hold Tank, Caustic Wash Tank And Caustic Storage Tank Samples From Modular Caustic-Side Solvent Extraction Unit During Macrobatch 6 Operations

Strip Effluent Hold Tank (SEHT), Decontaminated Salt Solution Hold Tank (DSSHT), Caustic Wash Tank (CWT) and Caustic Storage Tank (CST) samples from the Interim Salt Disposition Project (ISDP) Salt Batch (“Macrobatch”) 6 have been analyzed for 238 Pu, 90 Sr, 137 Cs, and by Inductively Coupled Plasma Emission Spectroscopy (ICPES). The Pu, Sr, and Cs results from the current Macrobatch 6 samples are similar to those from comparable samples in previous Macrobatch 5. In addition the SEHT and DSSHT heel samples (i.e. ‘preliminary’) have been analyzed and reported to meet NGS Demonstration Plan requirements. From a bulk chemical point of view, the ICPES results do not vary considerably between this and the previous samples. The titanium results in the DSSHT samples continue to indicate the presence of Ti, when the feed material does not have detectable levels. This most likely indicates that leaching of Ti from MST has increased in ARP at the higher free hydroxide concentrations in the current feed.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2014

Visual Sample Plan Version 7.0 User's Guide

User's guide for VSP 7.0 This user's guide describes Visual Sample Plan (VSP) Version 7.0 and provides instructions for using the software. VSP selects the appropriate number and location of environmental samples to ensure that the results of statistical tests performed to provide input to risk decisions have the required confidence and performance. VSP Version 7.0 provides sample-size equations or algorithms needed by specific statistical tests appropriate for specific environmental sampling objectives. It also provides data quality assessment and statistical analysis functions to support evaluation of the data and determine whether the data support decisions regarding sites suspected of contamination. The easy-to-use program is highly visual and graphic. VSP runs on personal computers with Microsoft Windows operating systems (XP, Vista, Windows 7, and Windows 8). Designed primarily for project managers and users without expertise in statistics, VSP is applicable to two- and three-dimensional populations to be sampled (e.g., rooms and buildings, surface soil, a defined layer of subsurface soil, water bodies, and other similar applications) for studies of environmental quality. VSP is also applicable for designing sampling plans for assessing chem/rad/bio threat and hazard identification within rooms and buildings, and for designing geophysical surveys for unexploded ordnance (UXO) identification.

97 MATHEMATICS AND COMPUTING↗

DOE OSTI2014

Analytical Results for MOX Colemanite Concrete Samples Received on January 15, 2013

The Mixed Oxide Fuel Fabrication Facility (MFFF) will use colemanite bearing concrete neutron absorber panels credited with attenuating neutron flux in the criticality design analyses and shielding operators from radiation. The Savannah River National Laboratory (SRNL) is tasked with measuring the total density, partial hydrogen density, and partial boron density of the colemanite concrete. SRNL received twelve samples of colemanite concrete for analysis on January 15, 2013. The average total density of each of the samples measured by the ASTM method C 642, the average partial hydrogen density was measured using method ASTM E 1131, and the average partial boron density of each sample was measured according to ASTM C 1301. The lower limits and measured values for the total density, hydrogen partial density, and boron partial density are presented. For all the samples tested, the total density and the hydrogen partial density met or exceeded the specified limit. All of the samples met or exceeded the boron partial density lower bound with the exception of samples G3-M11-2000-H, G3-M11-3000-M, and G5-M1-3000-H which are below the limit of 1.65E-01 g/cm 3 .

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

DOE OSTI2014

Assessment of the LV-S2 & LV-S3 Stack Sampling Probe Locations for Compliance with ANSI/HPS N13.1-1999

This document reports on a series of tests conducted to assess the proposed air sampling locations for the Hanford Tank Waste Treatment and Immobilization Plant (WTP) Group 1-2A exhaust stacks with respect to the applicable criteria regarding the placement of an air sampling probe. The LV-C2, LV-S2, and LV-S3 exhaust stacks were tested together as a group (Test Group 1-2A). This report only covers the results of LV-S2 and LV-S3; LV-C2 will be reported on separately. Federal regulations1 require that a sampling probe be located in the exhaust stack according to the criteria established by the American National Standards Institute/Health Physics Society (ANSI/HPS) N13.1-1999, Sampling and Monitoring Releases of Airborne Radioactive Substances from the Stack and Ducts of Nuclear Facilities. 2 These criteria address the capability of the sampling probe to extract a sample that represents the effluent stream.

Glissmeyer, John A.↗

DOE OSTI2015

Results of Hg Speciation Testing on 2Q15 Tank 50 and Tank 21 TCLP Extraction Fluid Samples and Tank 49 Material

The Savannah River National Laboratory (SRNL) was tasked with preparing and shipping samples for Hg speciation by Eurofins Frontier Global Sciences, Inc. in Seattle, WA on behalf of the Savannah River Remediation (SRR) Mercury Task Team. The sixth shipment of samples was designated to include SRNL Toxicity Characteristic Leaching Procedure (TCLP) extraction fluids from the 2Q15 Tank 50 and Tank 21 saltstone waste forms as well as Tank 49 material (HTF-49-15-45) collected on May 7, 2015. The TCLP extraction fluid samples were collected from duplicate extractions of two crushed saltstone fractions, one designated as “large” and one designated as “normal”. Both fractions of 2Q15 Tank 50 and Tank 21 saltstones (eight samples) were collected with the minimum possible free headspace and refrigerated prior to final dilution. The Tank 49 sample was left in its stainless steel dip bottle until it was opened and an aliquot diluted 1:100 with Eurofins deionized water on May 21, 2015. Following initial dilution in the Shielded Cells, the diluted Tank 49 sample was immediately moved to refrigeration and kept in the dark until prepared for shipment to Eurofins.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2016

Analysis Of Condensate Samples In Support Of The Antifoam Degradation Study

The degradation of Antifoam 747 to form flammable decomposition products has resulted in declaration of a Potential Inadequacy in the Safety Analysis (PISA) for the Defense Waste Processing Facility (DWPF). Savannah River National Laboratory (SRNL) testing with simulants showed that hexamethyldisiloxane (HMDSO), trimethylsilanol (TMS), and 1-propanal are formed in the offgas from the decomposition of the antifoam. A total of ten DWPF condensate samples from Batch 735 and 736 were analyzed by SRNL for three degradation products and additional analytes. All of the samples were analyzed to determine the concentrations of HMDSO, TMS, and propanal. The results of the organic analysis found concentrations for propanal and HMDSO near or below the detection limits for the analysis. The TMS concentrations ranged from below detection to 11 mg/L. The samples from Batch 736 were also analyzed for formate and oxalate anions, total organic carbon, and aluminum, iron, manganese, and silicon. Most of the samples contained low levels of formate and therefore low levels of organic carbon. These two values for each sample show reasonable agreement in most cases. Low levels of all the metals (Al, Fe, Mn, and Si) were present in most of the samples.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2016

ANALYSIS OF CONDENSATE SAMPLES IN SUPPORT OF THE ANTIFOAM DEGRADATION STUDY

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

DOE OSTI2016

Radiological results for samples collected on paired glass- and cellulose-fiber filters at the Sandia complex, Tonopah Test Range, Nevada

Airborne particulates are collected at U.S. Department of Energy sites that exhibit radiological contamination on the soil surface to help assess the potential for wind to transport radionuclides from the contamination sites. Collecting these samples was originally accomplished by drawing air through a cellulose-fiber filter. These filters were replaced with glass-fiber filters in March 2011. Airborne particulates were collected side by side on the two filter materials between May 2013 and May 2014. Comparisons of the sample mass and the radioactivity determinations for the side-by-side samples were undertaken to determine if the change in the filter medium produced significant results. The differences in the results obtained using the two filter types were assessed visually by evaluating the time series and correlation plots and statistically by conducting a nonparametric matched-pair sign test. Generally, the glass-fiber filters collect larger samples of particulates and produce higher radioactivity values for the gross alpha, gross beta, and gamma spectroscopy analyses. However, the correlation between the radioanalytical results for the glass-fiber filters and the cellulose-fiber filters was not strong enough to generate a linear regression function to estimate the glass-fiber filter sample results from the cellulose-fiber filter sample results.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

DOE OSTI2015

2015 Long-Term Hydrologic Monitoring Program Sampling and Analysis Results at Rio Blanco, Colorado

The U.S. Department of Energy (DOE) Office of Legacy Management conducted annual sampling at the Rio Blanco, Colorado, Site for the Long-Term Hydrologic Monitoring Program (LTHMP) on May 20–21, 2015. This report documents the analytical results of the Rio Blanco annual monitoring event, the trip report, and the data validation package. The groundwater and surface water monitoring samples were shipped to the GEL Group Inc. laboratories for conventional analysis of tritium and analysis of gamma-emitting radionuclides by high-resolution gamma spectrometry. A subset of water samples collected from wells near the Rio Blanco site was also sent to GEL Group Inc. for enriched tritium analysis. All requested analyses were successfully completed. Samples were collected from a total of four onsite wells, including two that are privately owned. Samples were also collected from two additional private wells at nearby locations and from nine surface water locations. Samples were analyzed for gamma-emitting radionuclides by high-resolution gamma spectrometry, and they were analyzed for tritium using the conventional method with a detection limit on the order of 400 picocuries per liter (pCi/L). Four locations (one well and three surface locations) were analyzed using the enriched tritium method, which has a detection limit on the order of 3 pCi/L. The enriched locations included the well at the Brennan Windmill and surface locations at CER-1, CER-4, and Fawn Creek 500 feet upstream.

54 ENVIRONMENTAL SCIENCES↗