Isotope Ratio Infrared Spectroscopy Research Articles

Organic contamination detection for isotopic analysis of water by laser spectroscopy.

Hydrogen and oxygen stable isotope ratios (δ2 H, δ17 O, and δ18 O values) are commonly used tracers of water. These ratios can be measured by isotope ratio infrared spectroscopy (IRIS). However, IRIS approaches are prone to errors induced by organic compounds present in plant, soil, and natural water samples. A novel approach using 17 O-excess values has shown promise for flagging spectrally contaminated plant samples during IRIS analysis. A systematic assessment of this flagging system is needed to prove it useful. Errors induced by methanol and ethanol water mixtures on measured IRIS and isotope ratio mass spectrometry (IRMS) results were evaluated. For IRIS analyses both liquid- and vapour-mode (via direct vapour equilibration) methods are used. The δ2 H, δ17 O, and δ18 O values were measured and compared with known reference values to determine the errors induced by methanol and ethanol contamination. In addition, the 17 O-excess contamination detection approach was tested. This is a post-processing detection tool for both liquid and vapour IRIS triple-isotope analyses, utilizing calculated 17 O-excess values to flag contaminated samples. Organic contamination induced significant errors in IRIS results, not seen in IRMS results. Methanol caused larger errors than ethanol. Results from vapour-IRIS analyses had larger errors than those from liquid-IRIS analyses. The 17 O-excess approach identified methanol driven error in liquid- and vapour-mode IRIS samples at levels where isotope results became unacceptably erroneous. For ethanol contaminated samples, a mix of erroneous and correct flagging occurred with the 17 O-excess method. Our results indicate that methanol is the more problematic contaminant for data corruption. The 17 O-excess method was therefore useful for data quality control. Organic contamination caused significant errors in IRIS stable isotope results. These errors were larger during vapour analyses than during liquid IRIS analyses, and larger for methanol than ethanol contamination. The 17 O-excess method is highly sensitive for detecting narrowband (methanol) contamination error in vapour and liquid analysis modes in IRIS.

Isotope ratio infrared spectroscopy analysis of water samples without memory effects.

Since their introduction more than a decade ago, isotope ratio infrared spectroscopy (IRIS) systems have rapidly become the standard for oxygen (δ18 O) and hydrogen (δ2 H) isotope analysis of water samples. An important disadvantage of IRIS systems is the well-documented sample-to-sample memory effect, which requires each sample to be analyzed multiple times before the desired accuracy is reached, lengthening analysis times and driving up the costs of analyses. We present an adapted set-up and calculation protocol for fully automated analysis of water samples using a Picarro L2140-i cavity ring-down spectroscopy instrument. The adaptation removes memory effects by use of a continuously moisturized nitrogen carrier gas. Water samples of 0.5 μL are measured on top of the water vapor background, after which isotope ratios are calculated by subtraction of the background from the sample peaks. With this new technique, single injections of water samples have internal precisions (1σ) below 0.05‰ for δ18 O values and 0.1‰ for δ2 H values, regardless of the isotope ratio of the previous sample. Precision is worse, however, when the isotope difference between the sample and background water is too large (i.e., exceeding approximately 9‰ for δ18 O values and 70‰ for δ2 H values). Isotope ratios show negligible drift across the four weeks within which the experiments were performed. The single-injection 1σ precision for 17 O excess (Δ'17 O) determined with this method is 60 per meg. Our experiments demonstrate that by removing sample-to-sample memory effects with a moisturized carrier gas, the time for measurement of δ18 O and δ2 H values using an IRIS system can be reduced markedly without compromising the analytical precision and accuracy. Thorough replication is needed to achieve sufficiently low uncertainties for Δ'17 O.

The influence of memory, sample size effects, and filter paper material on online laser-based plant and soil water isotope measurements.

The recent development of isotope ratio infrared spectroscopy (IRIS) was quickly followed by the addition of online extraction and analysis systems, making it faster and easier to measure soil and plant water isotopes. However, memory and sample size effects limit the efficiency and accuracy of these new setups. In response, this study presents a scheme dedicated to estimating and eliminating these two effects. Memory effect was determined by injecting two standard waters alternately. Each standard was injected nine times in a row and analyzed using induction module cavity ring-down spectroscopy (IM-CRDS). Memory coefficients were calculated using a new "multistage jump" algorithm. Sample size effects were evaluated by injecting water volumes ranging from 1μL to 6μL. Finally, the influence of cellulose filter paper on the isotopic measurements, the memory, and the sample size effect was evaluated by comparing it with glass filter paper. Memory effects were detected for both δ18 O and δ2 H values, with the latter being stronger. Isotopic differences between replicates of the same plant or soil sample showed a clear decrease after memory correction. A small water volume effect was found only when the injected water volume was larger than 3μL. However, while the correction method performed well for laboratory-made samples, it did not for field samples, due to the heterogeneity of the isotopic composition of the samples. Stronger memory and water volume effects were found for cellulose filter paper. The memory coefficients and the water volume-isotope relationship improved the consistency and accuracy of both laboratory and field data. Our results indicate that cellulose filter paper may not be a suitable medium to measure standard waters and evaluate memory and water volume effects. Finally, a detailed correction and calibration protocol is suggested, along with notes on best practices to obtain good-quality IM-CRDS data. Copyright © 2017 John Wiley & Sons, Ltd.

Intercomparison of two cavity ring-down spectroscopy analyzers for atmospheric <sup>13</sup>CO<sub>2</sub> ∕ <sup>12</sup>CO<sub>2</sub> measurement

Abstract. Isotope ratio infrared spectroscopy (IRIS) permits continuous in situ measurement of CO2 isotopic composition under ambient conditions. Previous studies have mainly focused on single IRIS instrument performance; few studies have considered the comparability among different IRIS instruments. In this study, we carried out laboratory and ambient measurements using two Picarro CO2δ13C analyzers (G1101-i and G2201-i (newer version)) and evaluated their performance and comparability. The best precision was 0.08–0.15 ‰ for G1101-i and 0.01–0.04 ‰ for G2201-i. The dependence of δ13C on CO2 concentration was 0.46 ‰ per 100 ppm and 0.09 ‰ per 100 ppm, the instrument drift ranged from 0.92–1.09 ‰ and 0.19–0.37 ‰, and the sensitivity of δ13C to the water vapor mixing ratio was 1.01 ‰ ∕ % H2O and 0.09 ‰ ∕ % H2O for G1101-i and G2201-i, respectively. The accuracy after correction by the two-point mixing ratio gain and offset calibration method ranged from −0.04–0.09 ‰ for G1101-i and −0.13–0.03 ‰ for G2201-i. The sensitivity of δ13C to the water vapor mixing ratio improved from 1.01 ‰ ∕ % H2O before the upgrade of G1101-i (G1101-i-original) to 0.15 ‰ ∕ % H2O after the upgrade of G1101-i (G1101-i-upgraded). Atmospheric δ13C measured by G1101-i and G2201-i captured the rapid changes in atmospheric δ13C signals on hourly to diurnal cycle scales, with a difference of 0.07 ± 0.24 ‰ between G1101-i-original and G2201-i and 0.05 ± 0.30 ‰ between G1101-i-upgraded and G2201-i. A significant linear correlation was observed between the δ13C difference of G1101-i-original and G2201-i and the water vapor concentration, but there was no significant correlation between the δ13C difference of G1101-i-upgraded and G2201-i and the water vapor concentration. The difference in the Keeling intercept values decreased from 1.24 ‰ between G1101-i-original and G2201-i to 0.36 ‰ between G1101-i-upgraded and G2201-i, which indicates the importance of consistency among different IRIS instruments.

Improved removal of volatile organic compounds for laser-based spectroscopy of water isotopes.

Volatile organic compounds (VOCs) such as methanol and ethanol in water extracted from plants cause spectral interference in isotope ratio infrared spectroscopy (IRIS). This contamination degrades the accuracy of measurements, limiting the use of IRIS. In response, this study presents a new decontamination method of VOCs for enhanced IRIS measurements. The isotopic compositions of water from laboratory-made and field-collected plant samples pre- and post-treatment were analyzed using IRIS. Traditional treatment methods of activated charcoal and commercial pre-combustion systems (MCM) were compared with our new treatment method that implements solid-phase extraction (SPE). The absolute concentrations of contaminants pre- and post-treatment were determined using (1)H and (13)C nuclear magnetic resonance to assess the effectiveness of the different treatments. SPE removes an average of 86.7% and 78.8% ethanol and methanol, respectively, significantly reducing spectral interference. SPE reduces errors to within instrumental noise for both ethanol and methanol at concentrations found in nature (<3.0% and 0.08%, respectively). Activated charcoal minimally affected alcohol concentrations. MCM significantly worsened ethanol-contaminated water isotope measurements by producing primary alcohol oxidation products such as formic acid, another compound that interferes with IRIS absorption. SPE is an effective, low-cost method for eliminating errors in ethanol-contaminated samples. For samples where methanol is prevalent, combining SPE and MCM is more effective than the use of SPE alone. Hence, SPE treatment alone or in conjunction with MCM is recommended as an effective pre-analysis purification method for water extracted from plants.

Isotope-ratio infrared spectroscopy: a reliable tool for the investigation of plant-water sources?

Stable isotopes are extensively used as tracers for the study of plant-water sources. Isotope-ratio infrared spectroscopy (IRIS) offers a cheaper alternative to isotope-ratio mass spectroscopy (IRMS), but its use in studying plant and soil water is limited by the spectral interference caused by organic contaminants. Here, we examine two approaches to cope with contaminated samples in IRIS: on-line oxidation of organic compounds (MCM) and post-processing correction. We assessed these methods compared to IRMS across 136 samples of xylem and soil water, and a set of ethanol- and methanol-water mixtures. A post-processing correction significantly improved IRIS accuracy in both natural samples and alcohol dilutions, being effective with concentrations up to 8% of ethanol and 0.4% of methanol. MCM outperformed the post-processing correction in removing methanol interference, but did not effectively remove interference for high concentrations of ethanol. By using both approaches, IRIS can overcome with reasonable accuracy the analytical uncertainties associated with most organic contaminants found in soil and xylem water. We recommend the post-processing correction as the first choice for analysis of samples of unknown contamination. Nevertheless, MCM can be more effective for evaluating samples containing contaminants responsible for strong spectral interferences at low concentrations, such as methanol.

Evaluating calibration strategies for isotope ratio infrared spectroscopy for atmospheric &amp;lt;sup&amp;gt;13&amp;lt;/sup&amp;gt;CO&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; / &amp;lt;sup&amp;gt;12&amp;lt;/sup&amp;gt;CO&amp;lt;sub&amp;gt;2&amp;lt;/sub&amp;gt; measurement

Abstract. Isotope ratio infrared spectroscopy (IRIS) provides an in situ technique for measuring δ13C in atmospheric CO2. A number of methods have been proposed for calibrating the IRIS measurements, but few studies have systematically evaluated their accuracy for atmospheric applications. In this study, we carried out laboratory and ambient measurements with two commercial IRIS analyzers and compared the accuracy of four calibration strategies. We found that calibration based on the 12C and 13C mixing ratios (Bowling et al., 2003) and on linear interpolation of the measured delta using the mixing ratio of the major isotopologue (Lee et al., 2005) yielded accuracy better than 0.06‰. Over a 7-day atmospheric measurement in Beijing, the two analyzers agreed to within −0.02 ± 0.18‰ after proper calibration. However, even after calibration the difference between the two analyzers showed a slight correlation with concentration, and this concentration dependence propagated through the Keeling analysis, resulting in a much larger difference of 2.44‰ for the Keeling intercept. The high sensitivity of the Keeling analysis to the concentration dependence underscores the challenge of IRIS for atmospheric research.

Isotopic composition of transpiration and rates of change in leaf water isotopologue storage in response to environmental variables

During daylight hours, the isotope composition of leaf water generally approximates steady-state leaf water isotope enrichment model predictions. However, until very recently there was little direct confirmation that isotopic steady-state (ISS) transpiration in fact exists. Using isotope ratio infrared spectroscopy (IRIS) and leaf gas exchange systems we evaluated the isotope composition of transpiration and the rate of change in leaf water isotopologue storage (isostorage) when leaves were exposed to variable environments. In doing so, we developed a method for controlling the absolute humidity entering the gas exchange cuvette for a wide range of concentrations without changing the isotope composition of water vapour. The measurement system allowed estimation of (18)O enrichment both at the evaporation site and for bulk leaf water, in the steady state and the non-steady state. We show that non-steady-state effects dominate the transpiration isoflux even when leaves are at physiological steady state. Our results suggest that a variable environment likely prevents ISS transpiration from being achieved and that this effect may be exacerbated by lengthy leaf water turnover times due to high leaf water contents.

Optimization of instrument setup and post‐run corrections for oxygen and hydrogen stable isotope measurements of water by isotope ratio infrared spectroscopy (IRIS)

Light stable isotope analyses of hydrogen (2H/1H) and oxygen (18O/16O) of water are used in many terrestrial and marine aquatic studies. The advantage of using stable isotope ratios is that water molecules serve as ubiquitous and already present natural tracers. Within recent years, these analyses have been revolutionized by the development of new isotope ratio laser spectroscopy (IRIS) systems that are cheaper, more robust, and mobile compared with traditional isotope ratio mass spectrometry (IRMS). Although easier to operate, laser systems also need thorough calibration with international reference materials, and raw data need correction for analytical effects (i.e., memory and drift). This study presents modifications to the hardware for liquid water injection, an optimized sequence layout and a simple post‐run correction procedure. These protocols will maximize precision, accuracy, and sample throughput via an efficient memory correction. The number of injections per unknown sample can be reduced to 4 or less. This procedure meets the demands of faster throughput with reduced costs per analysis. Procedures presented here are based on real analyses. They were also verified by an international proficiency test and traditional IRMS techniques.

Intercomparison of Four Commercial Analyzers for Water Vapor Isotope Measurement

Abstract The δ18O and δD of atmospheric water vapor are important tracers in hydrological and ecological studies. Isotope ratio infrared spectroscopy (IRIS) provides an in situ technology for measuring δ18O and δD in ambient conditions. An intercomparison experiment was carried out with four commercial IRIS analyzers to characterize their performance and transferability of calibration methods. Over a 15-day atmospheric measurement, during which the water vapor concentration ranged from 14 to 27 mol mol−1 and the isotopic ratios spanned about 90‰ and 13‰ for δD and δ18O, respectively, these analyzers tracked the natural variability in ambient conditions very well and achieved an average difference between one another within 2‰ for δD and within 0.1‰ for δ18O after calibration at appropriate frequencies. Two of the calibration methods (discrete liquid water injection and continuous dripping) agreed with each other within the tolerance thresholds of 2‰ for δD and 0.1‰ for δ18O. The Rayleigh distillation technique appeared to be acceptable as a calibration standard for δD but not for δ18O. The δD measurements were less prone to concentration dependence errors than the δ18O measurements. The concentration dependence underscores the importance of using a calibration procedure at multiple mixing ratios to bracket the range of natural variability.

Identification and correction of spectral contamination in 2H/1H and 18O/16O measured in leaf, stem, and soil water

Plant water extracts typically contain organic materials that may cause spectral interference when using isotope ratio infrared spectroscopy (IRIS), resulting in errors in the measured isotope ratios. Manufacturers of IRIS instruments have developed post-processing software to identify the degree of contamination in water samples, and potentially correct the isotope ratios of water with known contaminants. Here, the correction method proposed by an IRIS manufacturer, Los Gatos Research, Inc., was employed and the results were compared with those obtained from isotope ratio mass spectrometry (IRMS). Deionized water was spiked with methanol and ethanol to create correction curves for δ(18)O and δ(2)H. The contamination effects of different sample types (leaf, stem, soil) and different species from agricultural fields, grasslands, and forests were compared. The average corrections in leaf samples ranged from 0.35 to 15.73‰ for δ(2)H and 0.28 to 9.27‰ for δ(18)O. The average corrections in stem samples ranged from 1.17 to 13.70‰ for δ(2)H and 0.47 to 7.97‰ for δ(18)O. There was no contamination observed in soil water. Cleaning plant samples with activated charcoal had minimal effects on the degree of spectral contamination, reducing the corrections, by on average, 0.44‰ for δ(2)H and 0.25‰ for δ(18)O. The correction method eliminated the discrepancies between IRMS and IRIS for δ(18)O, and greatly reduced the discrepancies for δ(2)H. The mean differences in isotope ratios between IRMS and the corrected IRIS method were 0.18‰ for δ(18)O, and -3.39‰ for δ(2)H. The inability to create an ethanol correction curve for δ(2)H probably caused the larger discrepancies. We conclude that ethanol and methanol are the primary compounds causing interference in IRIS analyzers, and that each individual analyzer will probably require customized correction curves.

Detailed assessment of isotope ratio infrared spectroscopy and isotope ratio mass spectrometry for the stable isotope analysis of plant and soil waters

As an alternative to isotope ratio mass spectrometry (IRMS), the isotope ratio infrared spectroscopy (IRIS) approach has the advantage of low cost, continuous measurement and the capacity for field-based application for the analysis of the stable isotopes of water. Recent studies have indicated that there are potential issues of organic contamination of the spectral signal in the IRIS method, resulting in incorrect results for leaf samples. To gain a more thorough understanding of the effects of sample type (e.g., leaf, root, stem and soil), sample species, sampling time and climatic condition (dry vs. wet) on water isotope estimates using IRIS, we collected soil samples and plant components from a number of major species at a fine temporal resolution (every 2 h for 24-48 h) across three locations with different climatic conditions in the Heihe River Basin, China. The hydrogen and oxygen isotopic compositions of the extracted water from these samples were measured using both an IRMS and an IRIS instrument. The results show that the mean discrepancies between the IRMS and IRIS approaches for δ(18) O and δD, respectively, were: -5.6‰ and -75.7‰ for leaf water; -4.0‰ and -23.3‰ for stem water; -3.4‰ and -28.2‰ for root water; -0.5‰ and -6.7‰ for xylem water; -0.06‰ and -0.3‰ for xylem flow; and -0.1‰ and 0.3‰ for soil water. The order of the discrepancy was: leaf > stem ≈ root > xylem > xylem flow ≈ soil. In general, species of the same functional types (e.g., woody vs. herbaceous) within similar habitats showed similar deviations. For different functional types, the differences were large. Sampling at nighttime did not remove the observed deviations.

Spectral analysis software improves confidence in plant and soil water stable isotope analyses performed by isotope ratio infrared spectroscopy (IRIS)

Previous studies have demonstrated the potential for large errors to occur when analyzing waters containing organic contaminants using isotope ratio infrared spectroscopy (IRIS). In an attempt to address this problem, IRIS manufacturers now provide post-processing spectral analysis software capable of identifying samples with the types of spectral interference that compromises their stable isotope analysis. Here we report two independent tests of this post-processing spectral analysis software on two IRIS systems, OA-ICOS (Los Gatos Research Inc.) and WS-CRDS (Picarro Inc.). Following a similar methodology to a previous study, we cryogenically extracted plant leaf water and soil water and measured the δ(2)H and δ(18)O values of identical samples by isotope ratio mass spectrometry (IRMS) and IRIS. As an additional test, we analyzed plant stem waters and tap waters by IRMS and IRIS in an independent laboratory. For all tests we assumed that the IRMS value represented the "true" value against which we could compare the stable isotope results from the IRIS methods. Samples showing significant deviations from the IRMS value (>2σ) were considered to be contaminated and representative of spectral interference in the IRIS measurement. Over the two studies, 83% of plant species were considered contaminated on OA-ICOS and 58% on WS-CRDS. Post-analysis, spectra were analyzed using the manufacturer's spectral analysis software, in order to see if the software correctly identified contaminated samples. In our tests the software performed well, identifying all the samples with major errors. However, some false negatives indicate that user evaluation and testing of the software are necessary. Repeat sampling of plants showed considerable variation in the discrepancies between IRIS and IRMS. As such, we recommend that spectral analysis of IRIS data must be incorporated into standard post-processing routines. Furthermore, we suggest that the results from spectral analysis be included when reporting stable isotope data from IRIS.

Accuracy and precision of a laser-spectroscopy approach to the analysis of δ2H and δ18O in human urine

The isotope ratio analysis of body water often involves large sample numbers and lengthy sample processing. Here we demonstrate the ability of isotope ratio infrared spectroscopy (IRIS) to rapidly and accurately analyse the isotope ratios of water in urine. We analysed water extracted from human urine using traditional isotope ratio mass spectrometry (IRMS) and compared those values with IRIS-analysed extracted water and un-extracted urine. Regression analyses for δ2H and δ18O values between (1) extracted water analysed via IRMS and IRIS and (2) urine and extracted water analysed via IRIS were significant (R 2=0.99). These results indicate that cryogenic distillation of urine was not required for an accurate estimate of the isotopic composition of urine when using IRIS.

Discrepancies between isotope ratio infrared spectroscopy and isotope ratio mass spectrometry for the stable isotope analysis of plant and soil waters

The use of isotope ratio infrared spectroscopy (IRIS) for the stable hydrogen and oxygen isotope analysis of water is increasing. While IRIS has many advantages over traditional isotope ratio mass spectrometry (IRMS), it may also be prone to errors that do not impact upon IRMS analyses. Of particular concern is the potential for contaminants in the water sample to interfere with the spectroscopy, thus leading to erroneous stable isotope data. Water extracted from plant and soil samples may often contain organic contaminants. The extent to which contaminants may interfere with IRIS and thus impact upon data quality is presently unknown. We tested the performance of IRIS relative to IRMS for water extracted from 11 plant species and one organic soil horizon. IRIS deviated considerably from IRMS for over half of the samples tested, with deviations as large as 46 per thousand (delta(2)H) and 15.4 per thousand (delta(18)O) being measured. This effect was reduced somewhat by using activated charcoal to remove organics from the water; however, deviations as large as 35 per thousand (delta(2)H) and 11.8 per thousand (delta(18)O) were still measured for these cleaned samples. Interestingly, the use of activated charcoal to clean water samples had less effect than previously thought for IRMS analyses. Our data show that extreme caution is required when using IRIS to analyse water samples that may contain organic contaminants. We suggest that the development of new cleaning techniques for removing organic contaminants together with instrument-based software to flag potentially problematic samples are necessary to ensure accurate plant and soil water analyses using IRIS.