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At least 379 records · Page 21

Measuring residual accelerations in the Spacelab environment

A triglycine sulfate (TGS) crystal growth experiment carried out in Spacelab 3 flown onboard the Shuttle orbiter in 1985 is described. The TGS experiment was aimed at examining diffusion limited crystal growth by minimizing convection in the microgravity environment of the Spacelab. During the experiment tiny crystals were found floating free in the growth solution. Since the free crystals were in a viscous fluid, the 'noise' accelerations of the Shuttle were damped out. This made it possible to measure the constant gravitational acceleration by tracking the positions of these crystals. The velocities and accelerations obtained by these measurements are presented.

Witherow, William K.↗

Toward Guided Mutagenesis: Gaussian Process Regression Predicts MHC Class II Antigen Mutant Binding

Antigen-specific immunotherapies (ASI) require successful loading and presentation of antigen peptides into the major histocompatibility complex (MHC) binding cleft. One route of ASI design is to mutate native antigens for either stronger or weaker binding interaction to MHC. Exploring all possible mutations is costly both experimentally and computationally. To reduce experimental and computational expense, here we investigate the minimal amount of prior data required to accurately predict the relative binding affinity of point mutations for peptide-MHC class II (pMHCII) binding. Using data from different residue subsets, we interpolate pMHCII mutant binding affinities by Gaussian process (GP) regression of residue volume and hydrophobicity. We apply GP regression to an experimental data set from the Immune Epitope Database, and theoretical data sets from NetMHCIIpan and Free Energy Perturbation calculations. We find that GP regression can predict binding affinities of nine neutral residues from a six-residue subset with an average R 2 coefficient of determination value of 0.62 ± 0.04 (±95% CI), average error of 0.09 ± 0.01 kcal/mol (±95% CI), and with an receiver operating characteristic (ROC) AUC value of 0.92 for binary classification of enhanced or diminished binding affinity. Similarly, metrics increase to an R2 value of 0.69 ± 0.04, average error of 0.07 ± 0.01 kcal/mol, and an ROC AUC value of 0.94 for predicting seven neutral residues from an eight-residue subset. Our work finds that prediction is most accurate for neutral residues at anchor residue sites without register shift. This work holds relevance to predicting pMHCII binding and accelerating ASI design.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Low NOx heavy fuel combustor concept program. Phase 1: Combustion technology generation

The viability of low emission nitrogen oxide (NOx) gas turbine combustors for industrial and utility application. Thirteen different concepts were evolved and most were tested. Acceptable performance was demonstrated for four of the combustors using ERBS fuel and ultralow NOx emissions were obtained for lean catalytic combustion. Residual oil and coal derived liquids containing fuel bound nitrogen (FBN) were also used at test fuels, and it was shown that staged rich/lean combustion was effective in minimizing the conversion of FBN to NOx. The rich/lean concept was tested with both modular and integral combustors. While the ceramic lined modular configuration produced the best results, the advantages of the all metal integral burners make them candidates for future development. An example of scaling the laboratory sized combustor to a 100 MW size engine is included in the report as are recommendations for future work.

Lew, H. G.↗

Multigrid calculation of three-dimensional viscous cascade flows

A three-dimensional code for viscous cascade flow prediction has been developed. The space discretization uses a cell-centered scheme with eigenvalue scaling to weigh the artificial dissipation terms. Computational efficiency of a four-stage Runge-Kutta scheme is enhanced by using variable coefficients, implicit residual smoothing, and a full-multigrid method. The Baldwin-Lomax eddy-viscosity model is used for turbulence closure. A zonal, nonperiodic grid is used to minimize mesh distortion in and downstream of the throat region. Applications are presented for an annular vane with and without end wall contouring, and for a large-scale linear cascade. The calculation is validated by comparing with experiments and by studying grid dependency.

Arnone, A.↗

Enzymatic Laser-Induced Graphene Biosensor for Electrochemical Sensing of the Herbicide Glyphosate

Glyphosate is a globally applied herbicide yet it has been relatively undetectable in-field samples outside of gold-standard techniques. Its presumed nontoxicity toward humans has been contested by the International Agency for Research on Cancer, while it has been detected in farmers’ urine, surface waters and crop residues. Rapid, on-site detection of glyphosate is hindered by lack of field-deployable and easy-to-use sensors that circumvent sample transportation to limited laboratories that possess the equipment needed for detection. Herein, the flavoenzyme, glycine oxidase, immobilized on platinum-decorated laser-induced graphene (LIG) is used for selective detection of glyphosate as it is a substrate for GlyOx. The LIG platform provides a scaffold for enzyme attachment while maintaining the electronic and surface properties of graphene. The sensor exhibits a linear range of 10–260 µm, detection limit of 3.03 µm, and sensitivity of 0.991 nA µm -1 . The sensor shows minimal interference from the commonly used herbicides and insecticides: atrazine, 2,4-dichlorophenoxyacetic acid, dicamba, parathion-methyl, paraoxon-methyl, malathion, chlorpyrifos, thiamethoxam, clothianidin, and imidacloprid. Sensor function is further tested in complex river water and crop residue fluids, which validate this platform as a scalable, direct-write, and selective method of glyphosate detection for herbicide mapping and food analysis.

47 OTHER INSTRUMENTATION↗

Evaluation of the TOPEX/POSEIDON altimeter system over the Great Lakes

The TOPEX/POSEIDON altimeter measurment system is evaluated for the first 46 repeat cycles (September 23, 1992-December 23, 1992) using tracks over the Great Lakes. The temporal variations in lake level are removed from the altimeter measurements using in-situ lake level measurements, thus permitting the performance of the altimeter system to be assessed. For the NASA altimeter, the root-mean-square (RMS) scatter of the residuals is 3.95 cm using all the tracks over the lakes. However, some of the scatter in this result is probably due to lake tides or seiche, which can amount to a few centimeters amplitude near the ends of the lakes. When the seven best tracks are used, which cross the center of the lakes where tides/seiche effects are minimal, the RMS error is included to either 2.9 or 3.0 cm, depending on whether the Centre National d'Etudes Spatiales (CNES) or NASA orbit is used. This places an upper limit on the error budget of the altimeter system, excluding ocean tides and inverse barometer effect. There are several short-period variations in the residuals. The most pronounced is a 55-day period, with a 1-cm amplitude, which we believe is (at least in part) due to orbit error. When the model-derived wet tropospheric correction is substituted for the TOPEX microwave radiometer correction, the RMS error increases significantly, possibly resulting in an annual cycle of a few centimeters. Evaluation of the ionospheric correction indicates that the dual-frequency correction provides an average improvement of 0.85 cm over the Doppler orbitography and radiopositioning integrated by satellite (DORIS) correction. Although there are insufficient data to directly assess the CNES altimeter, the relative bias between the altimeters is estimated to be either -14.3 or -15.6 cm (NASA altimeter measuring short), depending on whether the DORIS or dual-frequency ionospheric correction is applied to the NASA altimeter.

Morris, Charles S.↗

Molecular Dynamics Simulation of Atomic Interactions in the Vancomycin Binding Site

Vancomycin is a glycopeptide antibiotic produced by Amycolaptopsis orientalis used to treat serious infections by Grampositive pathogens including methicillin-resistant Staphylococcus aureus. Vancomycin inhibits cell wall biosynthesis by targeting lipid II, which is the membrane-bound peptidoglycan precursor. The heptapeptide aglycon structure of vancomycin binds to the D-Ala-D-Ala of the pentapeptide stem structure in lipid II. The third residue of vancomycin aglycon is asparagine, which is not directly involved in the dipeptide binding. Nonetheless, asparagine plays a crucial role in substrate recognition, as the vancomycin analogue with asparagine substituted by aspartic acid (V D ) shows a reduction in antibacterial activities. To characterize the function of asparagine, binding of vancomycin and its aspartic-acid-substituted analogue V D to L-Lys-D-Ala-D-Ala and L-Lys-D-Ala-D-Lac was investigated using molecular dynamic simulations. Binding interactions were analyzed using root-mean-square deviation (RMSD), two-dimensional (2D) contour plots, hydrogen bond analysis, and free energy calculations of the complexes. The analysis shows that the aspartate substitution introduced a negative charge to the binding cleft of V D , which altered the aglycon conformation that minimized the repulsive lone pair interaction in the binding of a depsipeptide. Our findings provide new insight for the development of novel glycopeptide antibiotics against the emerging vancomycin-resistant pathogens by chemical modification at the third residue in vancomycin to improve its binding affinity to the D-Ala-D-Lac-terminated peptidoglycan in lipid II found in vancomycin-resistant enterococci and vancomycinresistant S. aureus.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Trellis complexity bounds for decoding linear block codes

We consider the problem of finding a trellis for a linear block code that minimizes one or more measures of trellis complexity. The domain of optimization may be different permutations of the same code or different codes with the same parameters. Constraints on trellises, including relationships between the minimal trellis of a code and that of the dual code, are used to derive bounds on complexity. We define a partial ordering on trellises: If a trellis is optimum with respect to this partial ordering, if has the desirable property that it simultaneously minimizes all of the complexity measures examined. We examine properties of such optimal trellises and give examples of optimal permutations of codes, most notably the (48,24,12) quadratic residue code.

Kiely, A. B.↗

Trellis Decoding Complexity of Linear Block Codes

We consider the problem of finding a trellis for a linear block code that minimizes one or more measures of trellis complexity. The domain of optimization may be different permutations of the same code, or different codes with the same parameters. Constraints on trellises, including relationships between the minimal trellis of a code and that of the dual code, are used to derive bounds on complexity. We define a partial ordering on trellises: if a trellis is optimum with respect to this partial ordering, it has the desirable property that it simultaneously minimizes all of the complexity measures examined. We examine properties of such optimal trellises and give examples of optimal permutations of codes, most notably the (48,24,12) quadratic residue code.

trellis decoding↗

Island scanning pattern optimization for residual deformation mitigation in laser powder bed fusion via sequential inherent strain method and sensitivity analysis

Laser powder bed fusion (L-PBF) has emerged as one of the mainstream additive manufacturing approaches for fabricating metal parts with complex geometries and intricate internal structures. However, large deformation associated with rapid heating and cooling can lead to build failure and requires post-processing which may increase manufacturing cost and prolong the production period. Here in this work, an island scanning pattern design method is proposed to optimize the scanning direction of each island in order to reduce part deformation after cutting off the build platform. The objective of this optimization is to minimize the upward bending of the part after sectioning, which allows the part deformation to satisfy the tolerance requirement or reduce the post heat treatment time. Inherent strain method is employed in the sequential finite element analysis consisting of layer-by-layer activations and sectioning for fast residual distortion prediction. Full sequential sensitivity analysis for the formulated optimization is provided to update the island scanning directions. To show the feasibility and effectiveness of the proposed method, the scanning patterns of a block structure and a connecting rod were designed and parts were fabricated using an open architecture L-PBF machine. The fabrication experiments demonstrated that the residual deformation of both parts fabricated by optimized scanning pattern can be reduced by over 50% compared to the initial scanning patterns, which demonstrate the effectiveness of the proposed method.

36 MATERIALS SCIENCE↗

Improved Method Being Developed for Surface Enhancement of Metallic Materials

Surface enhancement methods induce a layer of beneficial residual compressive stress to improve the impact (FOD) resistance and fatigue life of metallic materials. A traditional method of surface enhancement often used is shot peening, in which small steel spheres are repeatedly impinged on metallic surfaces. Shot peening is inexpensive and widely used, but the plastic deformation of 20 to 40 percent imparted by the impacts can be harmful. This plastic deformation can damage the microstructure, severely limiting the ductility and durability of the material near the surface. It has also been shown to promote accelerated relaxation of the beneficial compressive residual stresses at elevated temperatures. Low-plasticity burnishing (LPB) is being developed as an improved method for the surface enhancement of metallic materials. LPB is being investigated as a rapid, inexpensive surface enhancement method under NASA Small Business Innovation Research contracts NAS3-98034 and NAS3-99116, with supporting characterization work at NASA. Previously, roller burnishing had been employed to refine surface finish. This concept was adopted and then optimized as a means of producing a layer of compressive stress of high magnitude and depth, with minimal plastic deformation (ref. 1). A simplified diagram of the developed process is given in the following figure. A single pass of a smooth, free-rolling spherical ball under a normal force deforms the surface of the material in tension, creating a compressive layer of residual stress. The ball is supported in a fluid with sufficient pressure to lift the ball off the surface of the retaining spherical socket. The ball is only in mechanical contact with the surface of the material being burnished and is free to roll on the surface. This apparatus is designed to be mounted in the conventional lathes and vertical mills currently used to machine parts. The process has been successfully applied to nickel-base superalloys by a team from the NASA Glenn Research Center, Lambda Research, and METCUT Research, as supported by the NASA Small Business Innovation Research Phase I and II programs, the Ultra Safe program, and the Ultra- Efficient Engine Technology (UEET) Program.

Gabb, Timothy P.↗

CHARMM-GUI Drude prepper for molecular dynamics simulation using the classical Drude polarizable force field

Explicit treatment of electronic polarizability in empirical force fields (FFs) represents an extension over a traditional additive or pairwise FF and provides a more realistic model of the variations in electronic structure in condensed phase, macromolecular simulations. To facilitate utilization of the polarizable FF based on the classical Drude oscillator model, Drude Prepper has been developed in CHARMM-GUI. Drude Prepper ingests additive CHARMM protein structures file (PSF) and pre-equilibrated coordinates in CHARMM, PDB, or NAMD format, from which the molecular components of the system are identified. These include all residues and patches connecting those residues along with water, ions, and other solute molecules. This information is then used to construct the Drude FF-based PSF using molecular generation capabilities in CHARMM, followed by minimization and equilibration. In addition, inputs are generated for molecular dynamics (MD) simulations using CHARMM, GROMACS, NAMD, and OpenMM. Validation of the Drude Prepper protocol and inputs is performed through conversion and MD simulations of various heterogeneous systems that include proteins, nucleic acids, lipids, polysaccharides, and atomic ions using the aforementioned simulation packages. Stable simulations are obtained in all studied systems, including 5 μs simulation of ubiquitin, verifying the integrity of the generated Drude PSFs. Additionally, the ability of the Drude FF to model variations in electronic structure is shown through dipole moment analysis in selected systems. Finally, the capabilities and availability of Drude Prepper in CHARMM-GUI is anticipated to greatly facilitate the application of the Drude FF to a range of condensed phase, macromolecular systems.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Analysis of semivolatile organics in liquid radioactive residue using sorptive stir bar and solvent back-extraction

Current methods for semivolatiles analysis in radioactive samples can produce large volumes of radioactive solvent residue. A method utilizing stir-bar sorptive extraction has been explored in this work for its applicability to radioactive waste samples. This low solvent analytical method may accelerate remediation, minimize hazardous solvent waste, and reduce exposure risk to workers. Organic compounds (polyaromatic hydrocarbons, chlorinated aromatics, and phenolic compounds) were chosen as surrogates for common Liquid Waste System (LWS) contaminants at Savannah River Site (Aiken, SC). Stir-bar extraction parameters (extraction time, matrix modification, and effective pH range) and solvent back extraction parameters (solvent type, volume, and extraction time) were optimized experimentally for the chosen compounds. Affinity of the stir-bar extraction polymer to radionuclides Cs-137 and Am-241 was observed to determine radionuclide concentration effects. The stir-bar method achieved mean recovery of 100 ± 0.7% (1σ), relative to 114 ± 7% using solvent extraction, while reducing weekly method hands-on time by 93.4% and solvent volume consumption by 99.3%. Sensitivity was improved by 378% in simulated tank waste and 278% in real-world LWS matrix, relative to solvent extraction. This work has produced a safe and optimized method for the low solvent analysis of organics in legacy radioactive tank waste by stir-bar sorptive extraction.

GC-MS↗

Proteolytic dissection of Zab, the Z-DNA-binding domain of human ADAR1

Zalpha is a peptide motif that binds to Z-DNA with high affinity. This motif binds to alternating dC-dG sequences stabilized in the Z-conformation by means of bromination or supercoiling, but not to B-DNA. Zalpha is part of the N-terminal region of double-stranded RNA adenosine deaminase (ADAR1), a candidate enzyme for nuclear pre-mRNA editing in mammals. Zalpha is conserved in ADAR1 from many species; in each case, there is a second similar motif, Zbeta, separated from Zalpha by a more divergent linker. To investigate the structure-function relationship of Zalpha, its domain structure was studied by limited proteolysis. Proteolytic profiles indicated that Zalpha is part of a domain, Zab, of 229 amino acids (residues 133-361 in human ADAR1). This domain contains both Zalpha and Zbeta as well as a tandem repeat of a 49-amino acid linker module. Prolonged proteolysis revealed a minimal core domain of 77 amino acids (positions 133-209), containing only Zalpha, which is sufficient to bind left-handed Z-DNA; however, the substrate binding is strikingly different from that of Zab. The second motif, Zbeta, retains its structural integrity only in the context of Zab and does not bind Z-DNA as a separate entity. These results suggest that Zalpha and Zbeta act as a single bipartite domain. In the presence of substrate DNA, Zab becomes more resistant to proteases, suggesting that it adopts a more rigid structure when bound to its substrate, possibly with conformational changes in parts of the protein.

Non-NASA Center↗

In-Situ Reaction Monitoring for Lunar Applications Utilizing a Single Quadrupole Residual Gas Analyzer

With a revived focus to create a permanent presence on the moon and in preparation for future Mars exploration, it is imperative that all resources are utilized to their fullest potential.[1] In-Situ Resource Utilization (ISRU) will be critical for future mission success as it would enable independent operation of missions while minimally relying on the complex supply chain created between the Earth, moon and Mars. One of the most critical resources that have been identified for ISRU is the creation of liquid oxygen(LO2) for not only breathable air, but also for rocket propellant. On the moon and potentially Mars, one of the most O2 rich resources is in the presence of regolith.[2],[3] Although there are numerous minerals within regolith and various processes to extract O2,this is not the scope of this paper. Since the O2 extraction method can vary, a critical standardized analytical method is needed to verify O2(or oxygenated precursor compounds) for extraction efficiency and purity. Our group utilizes a modified consumer off-the-shelf (COTS), Residual Gas Analyzer (RGA) Single Quadrupole Mass Spectrometer (QMS). Due to the current field mission requirements, the gas analysis module will need to rely only on mass-to-charge ratios (m/z) and peak intensities to differentiate and quantitate targeted gas-phase reactants or products, i.e. carbon monoxide(CO), carbon dioxide (CO2), methane (CH4) and hydrogen (H2). However, due to solar wind implanted volatiles and various other compounds present in lunar regolith, such as nitrogen (N2), helium (He), and deuterium (D2), deconvolution and quantification of isobaric compounds becomes quite difficult. Specifically, the presence of N2 makes quantification of CO, an oxygenated precursor, difficult due to both compounds having a peak at m/z 28.

Mass Spectrometry↗

In-Situ Reaction Monitoring for Lunar Applications Utilizing a Single Quadrupole Residual Gas Analyzer

With a revived focus to create a permeant presence on the moon and in preparation for future Mars exploration, it is imperative that all re-sources are utilized to their fullest potential. In-Situ Resource Utilization (ISRU) will be critical for future mission success as it would enable independent operation of missions while minimally relying on the complex supply chain created between the Earth, moon and Mars. One of the most critical resources that have been identified for ISRU is the creation of liquid oxygen (O2) for not only breathable air, but also for rocket fuel. On the moon and potentially Mars, one of the most O2 rich resources is in the presence of regolith. Although there are numerous minerals within regolith and various processes to extract O2, this is not the scope of this paper. Since the O2 extraction method can vary, a critical standardized analytical method is needed to verify O2 (or oxygenated precursor compounds) for extraction efficiency and purity. Our group utilizes a modified consumer off-the-shelf (COTS), Residual Gas Analyzer (RGA) Single Quadrupole Mass Spectrometer (QMS). Due to the current field mission requirements, the gas analysis module will need to rely only on mass-to-charge ratios (m/z) and peak intensities to differentiate and quantitate targeted gas-phase reactants or products, i.e. carbon monoxide (CO), carbon dioxide (CO2), methane (CH4) and hydrogen (H2). However, due to solar wind implanted volatiles and various other compounds present in lunar regolith, such as nitrogen (N2), helium (He), and deuterium (D2), deconvolution and quantification of isobaric compounds becomes quite difficult. Specifically, the presence of N2 makes quantification of CO, an oxygenated precursor, difficult due to both compounds having a peak at m/z 28. Approaches to differentiate isobaric components: Initially, the solution to overcome isobaric overlap was to use isotopic ratios, particularly be-tween carbon-13 and nitrogen-15. Although limited literature discuss isotopic composition and attempt to quantify the ratios, variation is particularly high and limited to a few lunar regolith samples. The following were identified as potential approaches to quantify isobaric species, N2 and CO, using our RGA. Threshold Ionization. CO and N2 both have peaks at m/z 28, but only N2 is capable of producing a peak at m/z 7. While a N2 peak at m/z 7 is generally rare, instrument parameters can be optimized in order to maximize the peak intensity. Typically a standard ionization energy of 70 eV is used to monitor a batch process and would produce reliable results for a majority of the reactants or products that we are interested in quantifying. However, to analyze the rare molecular fragment of N2 (N2+ at m/z 7), a different ionization energy may be beneficial. Threshold Ionization Mass Spectrometry (TIMS) will be explored to optimize the N2+ peak at m/z 7 by varying parameters such as ionization energy and pressure. Therefore, a quantitative value at m/z 7 will be used to determine the product gas concentration of N2, which will be subtracted from the total concentration determined at m/z 28 enabling us to approximate the amount of CO. Secondary Detector (infrared gas detector, IR). Another approach to quantitate and differentiate be-tween isobaric components is to leverage alternative modes of detection. Unlike mass spectrometry, which generally uses a separation step prior to analysis (i.e. gas chromatography), IR produces energy that is specifically absorbed depending on the bond and configuration of the molecule. This technique could be leveraged since it is generally only active on non-symmetric molecules, i.e. CO. Utilizing concentrations of CO determined from another detector could help correct concentrations of CO obtained from the QMS. However, the inclusion of another detector adds additional mission requirements, such as power, software changes and avionics updates that add additional cost and schedule requirements.

mass spectrometry↗

Lunar Basins: New Evidence from Gravity for Impact-Formed Mascons

The prominent gravity highs (mascons) associated with uncompensated mass anomalies in lunar mare basins are a dramatic expression of the present-day rigidity of the lunar lithosphere. First discovered in Lunar Orbiter tracking data, these about 350-mGal gravity highs have been redetermined from the analysis of Clementine and historical tracking. These highs coincide with topographic lows, indicating nonisostatic support. One of the rediscoveries of this analysis is the encirclement of the highs by substantial negative anomalies over topographic highs. Recent gravity fields are providing the increased resolution necessary to determine the causes of this unique mascon signature. The compensation of the basin anomalies remains controversial. The mascon highs have long been interpreted as the result of mare loading, subsequent to the decay of residual stresses resulting from the impact. Substantially more mare fill is required to produce mascon highs than has been inferred on geological grounds, and the amount of near-surface mass deficit required to produce a gravity most exceeds bounds inferred from terrestrial examples. This problem is most acute for the youngest basin, Orientale. Recent gravity fields from Lunar Prospector have suggested mascon highs associated with nonmare basins such as Mendel-Rydberg, or minimally filled basins like Humboldtianum, further calling this explanation into question. We suggest that the mascon gravity signal is produced by a combination of crustal thickness changes, manifested by central mantle uplift, outward displacement of crust, and downward flexure of the lithosphere under mare loading. The mantle uplift is superisostatic, maintained by residual stresses resulting from the process of impact cratering and modification. In particular, the process of crater collapse and mantle rebound terminates abruptly, leaving the mantle plug in a non-equilibrium state, surrounded by a ring of thickened crust. Viscous relaxation over geological timescales has erased some but not all of the signature of the impact process. Mantle uplift inferred from gravity modeling is inversely correlated with age. While the oldest basins such as South Pole Aitken are mainly compensated isotatically, the younger basins appear to have been in a state of superisostatic loading prior to mare emplacement. If this is true, this places an important constraint on the impact process at basin scales. The idea that rebound of the transient crater via acoustic fluidization may freeze substantial stresses imcomplete to this day, may be tested by examining the gravity signatures of major basins on terrestrial bodies. The moon provides the clearest resolved examples to date, but uncertainty in gravity knowledge remains problematic.

Neumann, Gregory A.↗

Practical considerations for amino acid isotope analysis

Over the last few decades, isotopic analysis of amino acids at the compound- and position-specific levels has been rapidly advancing across diverse fields. As these techniques progress, evaluation of isotopic fractionation associated with sample workup is essential. This critical review of analytical methods through the lens of isotope geochemistry provides a benchmark for researchers across disciplines seeking to make compound- and position-specific amino acid isotope measurements. We focus on preparation, acid hydrolysis, clean-up, derivatization, separation, and C, H, N, and S isotope measurement. Despite substantial customizability across these steps, the following general recommendations should maximize recovery while minimizing isotopic fractionation. Samples should be freeze-dried and stored anoxically at ≤ –20 °C prior to conventional acid hydrolysis (6N HCl, 110 °C, 20–24 h, anoxic), which suffices for many residues. Both gas and liquid chromatographic (GC and LC, respectively) techniques are well-established and separate about 15 amino acids; LC bypasses the need for derivatization, while GC provides higher sensitivity. Furthermore, when derivatization is needed, n-acetyl and alkoxycarbonyl esters provide the most reproducible C isotope ratios. For compound-specific analyses, online GC–IRMS and LC–IRMS systems offer the easiest workflow, but EA–IRMS enables potential multi-element isotope analysis. Emerging techniques like high-resolution mass spectrometry are also promising for multi-element analysis and recover position-specific isotopic information. Looking forward to the next decade of innovation, isotope geochemists and ecologists can improve amino acid isotope analysis by focusing on streamlining multi-element analysis and standardizing calibration practices across laboratories.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗