Isotope Shifts Research Articles

Configurational entropy of ice XIX and its isotope effect

Ice XIX is a partly hydrogen-ordered polymorph related to disordered ice VI, similar to ice XV. We here investigate the order–order–disorder sequence ice XIX→ice XV→ice VI based on calorimetry at ambient pressure both for D2O and H2O-ice XIX. From these data we extract configurational entropy differences between ice XIX, ice XV and ice VI. This task is complex because, unlike for all other ices, the order–disorder transition from ice XIX to ice VI takes place in two steps via ice XV. Even more challenging, these two steps take place in an overlapping manner, so that careful separation of slow kinetics is necessary. This is evidenced best by changing the heating rate in calorimetry experiments: For fast heating experiments the second step, disordering of ice XV, is suppressed because the first step, formation of ice XV from ice XIX, is too slow. The transient state ice VI‡ that is initially produced upon ice XIX decay then does not have enough time to convert to ice XV, but remains disordered all along. In order to tackle the challenge to determine the entropy difference between ice XIX and VI as well as the entropy difference between ice XV and VI we employ two different approaches that allow assessing the impact of kinetics on the entropy change. “Single peak integration” defines a kinetically limited result, but “combined peak integration” allows estimation of the true thermodynamic values. Our best estimate for the true value shows ice XIX to be much more ordered than ice XV (25 ± 3% vs 9 ± 4% of the Pauling entropy). For D2Oice XIX samples we obtain 28% of order, but only when a small number of fast H-isotope defects are used. In the second part we use these results to estimate the location of the ice XIX phase boundary both for protiated and deuterated ice XIX. The initial Clapeyron slope at ambient pressure is determined from the combination of neutron powder diffraction volume differences and calorimetry entropy differences data to be 21 K GPa−1 with an order–disorder transition temperature To-d(0.0 GPa) = 103 ± 1 K. An in situ bracketing experiment at 1.8 GPa yields To-d(1.8 GPa) = 116 ± 3 K, i.e., the phase boundary slope flattens at higher pressures. These data allow us to determine the region of thermodynamic stability of ice XIX in the phase diagram and to explain the surprising isotope shift reversal at 1.6 GPa compared to 0.0 GPa, i.e., why D2O-ice XIX disorders at lower temperatures than H2O-ice XIX at 1.6 GPa, but at higher temperatures at ambient pressures.

Quartz Crystal Microbalance as a Holistic Detector for Quantifying Complex Organic Matrices during Liquid Chromatography: 2. Compound-Specific Isotope Analysis.

In carbon-compound-specific isotope analysis (carbon CSIA) of environmental micropollutants, purification of samples is often required to guarantee accurate measurements of a target compound. A companion paper has brought forward an innovative approach to couple a quartz crystal microbalance (QCM) with high-performance liquid chromatography (HPLC) for the online quantification of matrices during a gradient HPLC purification. This work investigates the benefit for isotope analysis of polar micropollutants typically present in environmental samples. Here, we studied the impact of the natural organic matter (NOM) on the isotopic integrity of model analytes and the suitability of the NOM-to-analyte ratio as a proxy for the sample purity. We further investigated limitations and enhancement of HPLC purification using QCM on C18 and C8 phases for single and multiple targets. Strong isotopic shifts of up to 3.3% toward the isotopic signature of NOM were observed for samples with an NOM-to-analyte ratio ≥10. Thanks to QCM, optimization of matrix removal of up to 99.8% of NOM was possible for late-eluting compounds. The efficiency of HPLC purification deteriorated when aiming for simultaneous purification of two or three compounds, leading to up to 2.5% less NOM removal. Our results suggest that one optimized HPLC purification can be achieved through systematic screening of 3 to 5 different gradients, thereby leading to a shift of the boundaries of accurate carbon CSIA by up to 2 orders of magnitude toward lower micropollutant concentrations.

Low Mo mobility during the laterization of ultramafic bedrock: Evidence from the East Sulawesi Ophiolite, Indonesia

Mo (isotope) cycling during the chemical weathering of ultramafic bedrock remains poorly quantified, mainly as a result of the analytical challenges caused by low Mo concentration and complex matrix effects in these rock types. Here, we utilize an improved chemical separation protocol that enables the extraction of Mo while reducing Ru and Fe matrix effects. We apply this method to lateritic weathering profiles developed over ultramafic bedrock in a high-intensity tropical weathering regime. The Mo concentrations in the laterite samples are higher (0.022 to 0.58 μg·g−1) than those of the peridotite bedrock (0.006 to 0.021 μg·g−1). The concentration-weighted average δ98Mo of the laterite profiles is −0.05‰ (n = 17), which is slightly higher but very close to the average δ98Mo of the peridotite bedrock (0.17 ± 0.21‰ (2SD; n = 5). Weakly-laterized samples show somewhat low δ98Mo with minimum of −1.03‰ and a Δ98Molaterite-bedrock up to −0.86‰, possibly as a result of preferential adsorption of liberated light Mo onto Fe (oxyhydr)oxides. In contrast, strongly-laterized samples show an overall Mo concentration gain and a slight isotopic shift towards higher bulk δ98Mo, with a maximum δ98Mo of +0.12‰ and Δ98Molaterite-bedrock up to +0.42‰. This likely reflects the re-scavenging of Mo released from weakly-laterized horizons to the ubiquitous Fe (oxyhydr)oxides, with potential superimposition of additional heavy Mo from atmospheric and/or groundwater input. Overall, this suggests a small contribution of dissolved Mo derived from ultramafic bedrock weathering in tropical settings to the aquatic environment.

Where are the feeder channels for platinum reefs in the Bushveld Complex?

AbstractThe Bushveld Complex in South Africa hosts the lion’s share of the world’s noble metal resources in platinum reefs – thin layers of silicate/chromite rocks containing platinum-rich sulphides. The reefs are widely attributed to multiple replenishments by ore-forming magmas that have been entering the evolving Bushveld chamber through numerous feeder conduits. The replenishment events are marked by regional and local disconformities/unconformities, significant isotopic shifts, and notable reversals in the whole-rock and mineral compositions. Surprisingly, however, so far no single feeder conduit for platinum reefs has been found despite extensive surface and underground mining for over a century. Feeder conduits appear entirely absent from the Bushveld Complex. This paradox has long been known but has never been specifically addressed. Here, we suggest that the absence of feeder channels is a natural consequence of the magma chamber replenishment through a cumulate pile. The fossilization of the feeder channels in the cumulate pile is likely impeded by two principal factors: (a) a cumulate pile is too hot to enable efficient cooling and crystallization of magma flowing through the channels, and (b) the channels are closed by an adjacent elastically deformable pile immediately after cessation of the magma emplacement. The feeding dykes are thus absent because there is little chance for the conduits to get preserved in a hot and deformable cumulate pile of layered intrusions.

Spectroscopic study of the 4f76s2 8 S7/2∘ – 4f7(8S°) 6s6p(1P°) 8P5/2,7/2 transitions in neutral europium-151 and europium-153: absolute frequency and hyperfine structure

We report on spectroscopic measurements on the 4f76s28S7/2∘−4f7(8S∘)6s6p(1P∘)8P5/2,7/2 transitions at 466.32 nm and 462.85 nm, respectively, in neutral europium-151 and europium-153. The center of gravity frequencies for the 151 and 153 isotopes for both transitions are reported for the first time using saturated absorption spectroscopy. For the 6s6p(1P∘)8P5/2 state, the center of gravity frequencies were found to be 642,894,493.3(4) MHz and 642,891,693.3(9) MHz for the 151 and 153 isotopes, respectively. The hyperfine constants for the upper state were found to be A(151)=−157.01(3)MHz, B(151)=74.5(4)MHz and A(153)=−69.43(14)MHz, B(153)=191.0(26)MHz. These hyperfine values are all consistent with previously published results except for B(151) that has a small discrepancy. The isotope shift was found to be 2799.54(20) MHz, a small discrepancy with previously published results. For the 6s6p(1P∘)8P7/2 state, the center of gravity frequencies were found to be 647,708,930.6(6) MHz and 647,705,958.4(26) MHz for the 151 and 153 isotopes, respectively. The hyperfine constants for the upper state were found to be A(151)=−218.66(4)MHz, B(151)=−293.4(8)MHz and A(153)=−97.15(13)MHz, B(153)=−750(3)MHz. These values are all consistent with previously published results except for A(151) that has a small discrepancy. The isotope shift was found to be 2972.8(5) MHz, a small discrepancy with previously measured results.

Kink of the nuclear charge radius isotope shift and overlaps of the neutron and proton orbitals in lead

Within the relativistic mean field approximation, we analyse the kink effect (KE) in the evolution of the charge radius isotope shift of lead isotopes as a function of the neutron number N. We show that if the interactions between neutron and proton states responsible for the KE are assumed to be proportional either to the overlaps of their corresponding wave functions or to those of their corresponding probability density distributions, it is not possible, by themselves, to explain the KE. However, we find that the small component of the single-particle Dirac spinors plays a relevant role in the kink formation. By considering the contribution of the N−126 valence neutrons to the proton central potential, we can explain the generation of the KE and why neutrons in the 1i11/2 orbital are more kinky than when they are in the 2g9/2 orbital.

Isolating the Contributions from Moments of Inertia in Isotopic Shifts Measured by High-Resolution Cyclic Ion Mobility Separations.

The unexpected finding that isotopomers (i.e., isotopic isomers) can be separated with high-resolution ion mobility spectrometry-mass spectrometry (IMS-MS) has raised new structural considerations affecting an ion's mobility, namely its center of mass (CoM) and moments of inertia (MoI). Unfortunately, thus far, no studies have attempted to experimentally isolate either CoM or MoI, as they are intrinsically linked by their definitions, where MoI is calculated in relation to CoM. In this study, we designed and synthesized four isotopically labeled tetrapropylammonium (TAA3) ions, each with a unique mass distribution. Three of the synthesized TAA3 ions were labeled symmetrically, thus having identical CoM but differing MoI, which we verified using density functional theory (DFT) calculations. Consequently, we were able to isolate the effect of MoI changes in high-resolution IMS-MS separations. Cyclic ion mobility spectrometry-mass spectrometry (cIMS-MS) separations of the isotopically labeled TAA3 variants revealed isotopic mobility shifts attributable solely to changes in MoI. A 60-m cIMS-MS separation demonstrated that two nominally isobaric TAA3 pseudoisotopomers could be partially resolved, showcasing potential feasibility for isotopomer separations on commercially available IMS-MS platforms. With our previously established collision cross section (CCS) calibration protocol, we also quantified the relationship between MoI and CCS. Our results represent the first demonstration of IMS-MS separations based solely on MoI differences. We believe these findings will contribute important evidence to the growing body of literature on the physical nature of isotopic shifts in IMS-MS separations and work toward more accurate CCS predictions.

Spectroscopy of the 5s5p3P0→5s5d3D1 transition of strontium using laser cooled atoms

This article presents spectroscopy results of the 5s5p3P0→5s5d3D1 transition in all isotopes of laser cooled Sr atoms and the utility of this transition for repumping application. By employing the 5s5p3P0→5s5d3D1 (483 nm) transition in combination with the excitation of 5s5p3P2→5s6s3S1 (707 nm) transition, we observe a significant increase (∼13 fold) in the steady state number of atoms in the magneto-optic trap. This enhancement is attributed to the efficient repumping of Sr atoms that have decayed into the dark 5s5p3P2 state by returning them to the ground state 5s21S0 without any loss into the other states. The absolute transition frequencies were measured with an absolute accuracy of 30 MHz. To support our measurements, we performed Fock-space relativistic coupled-cluster calculations of the relevant parameters in Sr To further increase the accuracy of the calculated properties, corrections from the Breit, quantum electrodynamical and perturbative triples were also included. The calculated branching ratio for the repumping state confirms the significantly increased population in the 3P1 state. Thereby, leading to an increase of population of atoms trapped due to the enhanced repumping. Our calculated hyperfine-splitting energies are in excellent agreement with the measured values. Moreover, our calculated isotope shifts in the transition frequencies are in good agreement with our measured values.

The Toarcian Posidonia Shale at Salem (North Alpine Foreland Basin; South Germany): hydrocarbon potential and paleogeography

AbstractThe Posidonia Shale in the basement of the North Alpine Foreland Basin of southwestern Germany represents an important archive for environmental changes during the Toarcian oceanic anoxic event and the associated carbon isotope excursion (T-CIE). It is also an important hydrocarbon source rock. In the Salem borehole, the Posidonia Shale is ~ 10 m thick. The lower 7.5 m (1763.5–1756.0 m) of the Posidonian Shale and the uppermost part of the underlying Amaltheenton Formation were cored and studied using a total of 62 samples. Rock–Eval, palynological, maceral, biomarker and carbon isotope data were collected to assess variations in environmental conditions and to quantify the source rock potential. In contrast to most other Toarcian sections in southwest Germany, TOC contents are high in sediments deposited during the T-CIE, but reach a peak in post-CIE sediments. Biomarker ratios suggest that this reflects strong oxygen-depletion during the T-CIE (elegantulum to lower elegans subzones), but also during the falciferum Subzone, which is also reflected by a prolonged dinoflagellate cyst blackout. While sediments of the tenuicostatum Zone to the elegans Subzone are thinner than in neighbouring sections (e.g., Dotternhausen), sediments of the falciferum Subzone are unusually thick, suggesting that increased subsidence might have contributed to anoxia. The T-CIE interval is very thin (0.75 m). δ13C values of n-alkanes show that the maximum negative isotope shift predates the strongest basin restriction during the T-CIE and that the carbon isotope shift is recorded earlier for aquatic than for terrigenous organisms. In Salem, the Posidonia Shale is thermally mature and highly oil-prone. The residual source petroleum potential is about 0.8 tHC/m2. Graphical Abstract

Brief and intensive volcanic emissions from Ontong Java Nui heralded Oceanic Anoxic Event 1a

The volcanic episode that formed Ontong Java Nui (OJN) in the western Pacific ~ 120 million-year-ago is thought to have triggered Oceanic Anoxic Event (OAE) 1a, yet the cause-effect relationship remains insufficiently understood. Here, we present a Pb-Os-C isotope dataset for tracking OJN volcanism across a sedimentary sequence containing OAE1a record in the central Pacific. Lead isotopic evidence strongly supports the deposition of OJN-sourced volcanic ash layers at this site after a paired seawater δ13Ccarb–187Os/188Os shift and before the onset of OAE1a. The δ13Ccarb–187Os/188Os shift is attributable to the signature of the initial submarine OJN eruption. The subsequent unradiogenic Pb isotope shifts could represent ash fallout from the brief, explosive subaerial/shallow-marine eruption phase accompanied by intensive volcanic degassing and emissions. The apparent time lag between explosive OJN volcanism and OAE1a is intriguing, which can be accounted for by the delayed increase in the productivity after gas emissions.

Isotopic variations of Sm, Gd, Er and Yb found in planetary materials caused by neutron-capture reactions in nature

The isotopic shifts of 149Sm–150Sm and 157Gd–158Gd have often been observed in meteorites and lunar surface materials, because they result from the neutron-capture reactions associated with secondary neutrons produced by cosmic-ray irradiation. While the Sm and Gd isotopic shifts can mainly be used for the estimation of thermal neutron fluences that of 167Er–168Er has recently been applied in the estimation of epithermal neutron fluences. The systematic isotopic dataset of Sm, Gd and Er helps us to consider the details of planetary materials’ cosmic-ray exposure conditions using the balance of the fluences between thermal and epithermal neutrons. This paper reviews a series of isotopic variations of Sm, Gd, and Er in association with neutron-capture reactions for the application of planetary sciences. As a new attempt and possibility for better understanding the neutron fluence and its energy distribution, the use of Yb isotopic variation is then discussed using two different data sources, namely lunar regolith and the Oklo natural reactors. Finally, the preliminary result for the precise isotopic measurement of Yb is presented from the viewpoint of chemical separation and instrumental improvement.

The infrared absorption spectrum of phenylacetylene and its deuterated isotopologue in the mid- to far-IR.

Anharmonicity strongly influences the absorption and emission spectra of polycyclic aromatic hydrocarbon (PAH) molecules. Here, IR-UV ion-dip spectroscopy experiments together with detailed anharmonic computations reveal the presence of fundamental, overtone, as well as 2- and 3-quanta combination band transitions in the far- and mid-infrared absorption spectra of phenylacetylene and its singly deuterated isotopologue. Strong absorption features in the 400-900 cm-1 range originate from CH(D) in-plane and out-of-plane wags and bends, as well as bending motions including the C≡C and CH bonds of the acetylene substituent and the aromatic ring. For phenylacetylene, every absorption feature is assigned either directly or indirectly to a single or multiple vibrational mode(s). The measured spectrum is dense, broad, and structureless in many regions but well characterized by computations. Upon deuteration, large isotopic shifts are observed. At frequencies above 1500 cm-1 for d1-phenylacetylene, a one-to-one match is seen when comparing computations and experiments with all features assigned to combination bands and overtones. The C≡C stretch observed in phenylacetylene is not observed in d1-phenylacetylene due to a computed 40-fold drop in intensity. Overall, a careful treatment of anharmonicity that includes 2- and 3-quanta modes is found to be crucial to understand the rich details of the infrared spectrum of phenylacetylene. Based on these results, it can be expected that such an all-inclusive anharmonic treatment will also be key for unraveling the infrared spectra of PAHs in general.

Quantum symmetrization transition in superconducting sulfur hydride from quantum Monte Carlo and path integral molecular dynamics

AbstractWe study the structural phase transition, originally associated with the highest superconducting critical temperature Tc measured in high-pressure sulfur hydride. A quantitative description of its pressure dependence has been elusive for any ab initio theory attempted so far, raising questions on the actual mechanism leading to the maximum of Tc. Here, we estimate the critical pressure of the hydrogen bond symmetrization in the Im$$\bar{3}$$ 3 ¯ m structure, by combining density functional theory and quantum Monte Carlo simulations for electrons with path integral molecular dynamics for quantum nuclei. We find that the Tc maximum corresponds to pressures where local dipole moments dynamically form on the hydrogen sites, as precursors of the ferroelectric Im$$\bar{3}$$ 3 ¯ m-R3m transition, happening at lower pressures. For comparison, we also apply the self-consistent harmonic approximation, whose ferroelectric critical pressure lies in between the ferroelectric transition estimated by path integral molecular dynamics and the local dipole formation. Nuclear quantum effects play a major role in a significant reduction (≈50 GPa) of the classical ferroelectric transition pressure at 200 K and in a large isotope shift (≈25 GPa) upon hydrogen-to-deuterium substitution of the local dipole formation pressure, in agreement with the corresponding change in the Tc maximum location.

Isotopic shifts in 3 P states of the carbon atom

ABSTRACT Isotope shifts in the transition energies between several lowest 3 P e states of the carbon atom are calculated at the nonrelativistic level of theory. We considered the 12 C, 13 C, and 14 C isotopes, for which we performed variational calculations by expanding the wave functions of the atomic states in terms of all-electron explicitly correlated Gaussian functions. The Born–Oppenheimer approximation was not assumed in the calculations. By combining the computed isotope shifts with the experimentally derived Ritz wavelengths for the natural mixture of the isotopes we make predictions of the positions of the spectral lines for specific carbon isotopes. These predictions may be useful for guiding future experimental measurements of these lines using high-resolution spectroscopy.

Designer spectrographs for applications in the advanced undergraduate instructional lab

In an advanced undergraduate instructional laboratory, it is often necessary to analyze the spectrum of light emitted from an experimental setup. There are numerous instruments that are used to accomplish this analysis, including spectrometers and spectrographs. In this report, we present 3D-printed, low-budget spectrographs (∼ US $200), which are specifically designed for different applications. For example, one can either observe a visible spectrum over a large range of wavelengths about a desired center wavelength or achieve more precise measurements by choosing a smaller part of the visible spectrum. Our generalized design approach is well within the knowledge base of advanced undergraduate physics majors and can be applied to a wide range of applications within the visible spectrum. To demonstrate the utility of these designer spectrographs, we provide examples of recording multiple doublets in the sodium spectrum (a determination of the fine structure spin-orbit splitting of the 3p energy level) as well as measuring the wavelength differences between the hydrogen and deuterium Balmer alpha lines (a measurement of an isotope shift).

Isotopic effect of oxygen on the Raman mapping of a polycrystalline uranium dioxide UO2

AbstractUranium dioxide (UO2) is a widely studied material due to its use as fuel in pressurised water reactors (PWR), and Raman spectroscopy is a technique of choice to characterise the evolution of its microstructure. UO2 crystallises in a fluorite CaF2 (space group Fm‐3m) structure that gives rise to a unique Raman signature, the T2g band. However, several other bands are often detected whose attribution remains unclear. The present study gives new insights on the Raman spectrum of UO2 thanks to the combination of isotopic labelling with 18O and Raman imaging. In addition to the expected T2g, U2 (LO), 2LO and U3 bands, we have detected a doublet at 885 and 925 cm−1, a U* band at 555 cm−1 in some specific areas and two bands located at 367 and 1196 cm−1. All Raman bands shift under the effect of the replacement of 16O by 18O, excepting for the U* band that could not be detected anymore. The isotopic shift ratio could be determined for 20% and 30% 18O labelling. No discrepancy in band position is observed between grains and grain boundaries, except for the U2(LO) band. We also evidence a difference between the U defect bands and the 885 and 925 cm−1 doublet bands evolution under labelling, although the latter also seems to be connected to the presence of defects in the material.

Isotopic Shifts of Vibrational-Rotational Lines of SO2

Isotopic Shifts of Vibrational-Rotational Lines of SO2

Estimating Riparian Water Movement in an Incipient Karst Landscape—Minnesota, USA

Understanding rain, ground, and surface water interactions in riparian zones is essential for hydrologic and environmental management. The novelty of this study lies in its revelation of isotopic shifts and consequent evaluation of hydrologic pathways and processes within the forested riparian zones of three distinct streams, valleys, and riparian systems in a carbonate-sandstone incipient karst landscape. We collected water samples from three southeastern Minnesota catchments: Trout Brook, Crystal Creek, and Bridge Creek. A Picarro L2130i was used to measure δ18O for oxygen, and δD for deuterium in units of ‰ (per mil). We estimated the global meteoric line for the study sites, built upon aquifer age dating, and explored aquifer transit time for the study sites using a simple seasonal amplitude model. The results showed small amplitudes for 2020 and 2021, suggesting that bedrock aquifer water was the primary source water with a mean core transit time greater than 10 years. All three catchments were different but had similar bedrock valley types and riparian sediment. The primary driving factor was the seasonal precipitation input mixing with existing water. In a normal to wetter year, the isotopic data showed larger amplitude shifts between seasons with trendlines that adjusted depending on the temperature of the new water additions. The proposed approach is valuable in revealing complex hydrologic processes and pathways and can contribute extensively to the planning and management of karst riparian systems.

Measuring Hydroxyl Exchange Rates in Glycans Using a Synergistic Combination of Saturation Transfer and Relaxation Dispersion NMR.

Molecular recognition events mediated by glycans play pivotal roles in controlling the fate of diverse biological processes such as cellular communication and the immune response. The affinity of glycans for their target receptors is governed primarily by the hydrogen bonds formed by hydroxyl groups decorating the glycan surface. Hydroxyl exchange rate constants are therefore vital parameters that report on glycan structure and dynamics. Here we present a strategy for characterizing hydroxyl hydrogen/deuterium (H/D) exchange in glycans that employs a synergistic combination of 13C chemical exchange saturation transfer (CEST) and Carr-Purcell-Meiboom-Gill relaxation dispersion (CPMG) NMR methods. We show that the combination of CEST and CPMG experiments facilitates the sensitive detection of the small (∼0.1 ppm) two-bond deuterium isotope shift on a 13C nucleus when the attached hydroxyl group fluctuates between protonated and deuterated states. This shift is leveraged for measuring site-specific kinetic H/D exchange rate constants as well as thermodynamic free energies of isotope fractionation. The CEST and CPMG modules are integrated with a selective J-cross-polarization scheme that provides the flexibility for rapid characterization of H/D exchange at a specific hydroxyl site. Moreover, our approach enables the precise isothermal measurement of hydroxyl exchange rate constants without the need for cumbersome isotope labeling. The H/D exchange rate constants of three different glycans assessed using this method highlight its potential for detecting transient intra- and intermolecular hydrogen bonds. In addition, the trends in H/D exchange rate constants establish site-specific steric accessibility as a key determinant of solvent exchange dynamics in glycans.

Metal-binding amino acid ligands commonly found in metalloproteins differentially fractionate copper isotopes

Copper (Cu) is a cofactor in numerous key proteins and, thus, an essential element for life. In biological systems, Cu isotope abundances shift with metabolic and homeostatic state. However, the mechanisms underpinning these isotopic shifts remain poorly understood, hampering use of Cu isotopes as biomarkers. Computational predictions suggest that isotope fractionation occurs when proteins bind Cu, with the magnitude of this effect dependent on the identity and arrangement of the coordinating amino acids. This study sought to constrain equilibrium isotope fractionation values for Cu bound by common amino acids at protein metal-binding sites. Free and bound metal ions were separated via Donnan dialysis using a cation-permeable membrane. Isotope ratios of pre- and post-dialysis solutions were measured by MC-ICP-MS following purification. Sulfur ligands (cysteine) preferentially bound the light isotope (63Cu) relative to water (Δ65Cucomplex-free = − 0.48 ± 0.18‰) while oxygen ligands favored the heavy isotope (65Cu; + 0.26 ± 0.04‰ for glutamate and + 0.16 ± 0.10‰ for aspartate). Binding by nitrogen ligands (histidine) imparted no isotope effect (− 0.01 ± 0.04‰). This experimental work unequivocally demonstrates that amino acids differentially fractionate Cu isotopes and supports the hypothesis that metalloprotein biosynthesis affects the distribution of transition metal isotopes in biological systems.