Ratio Mass Spectrometry System Research Articles

Long-term performance of simultaneous measurement of stable isotopes of oxygen and carbon in cellulose with a high-temperature pyrolysis/gas chromatography/isotope ratio mass spectrometry system at the Institute for Geological and Geochemical Research

AbstractA high-temperature pyrolysis/gas chromatography/isotope ratio mass spectrometry system was established at the Institute for Geological and Geochemical Research in 2013. A dedicated field of application of the system is the simultaneous measurement of stable carbon and oxygen isotope ratios in the cellulose of modern, relict and subfossil plant tissues and sediments. The measurement protocol was fine-tuned during the first year of operation and documented in detail in this report. To quantify the long-term reproducibility of the simultaneous measurement of stable isotopes of oxygen and carbon in cellulose, a 2σ range inferred from repeated measurements of a Quality Assurance standard can be recommended: 0.16 and 0.20‰, for δ13C and δ18O, respectively. An extensive set of samples with known pyrolysis-based δ13C data was analyzed in combustion mode and the paired results were used to assess the necessity of adjustment of the pyrolysis-based δ13C measurements. The variances of the two datasets were not significantly different; the slope (intercept) of the regression was indistinguishable from unity (zero), suggesting that probably owing to the relatively frequent cleaning of the pyrolysis furnace, pyrolysis-based δ13C data neither suffer from a variance bias nor require a specific adjustment.

Evaluation of a quick one-step sample preparation method for the determination of the isotopic fingerprint of rapeseed (Brassica napus): Investigation of the influence of the use of 2,2-dimethoxypropane on compound-specific stable carbon and hydrogen isotope analyses by gas chromatography combustion/pyrolysis isotope ratio mass spectrometry.

Gas chromatographic analyses for vegetable oils require transesterification, which generally involves multiple steps, mainly to generate fatty acid methyl esters (FAMEs). A quick method based on acid-catalyzed transesterification using 2,2-dimethoxypropane (DMP) enables the conversion in one step, in a single reactor. For compound-specific stable carbon and hydrogen isotope analyses (C- and H-CSIA) of individual fatty acids (FAs) in oil, the verification of this one-step method has not yet been reported. In this study, we evaluated the feasibility of the one-step method for C- and H-CSIA of individual FAMEs in rapeseed samples. The focus was on the investigation of the influence of methanol, which was produced from the reactions of DMP with glycerol and water during transesterification, on the accuracy of isotope composition of FAMEs, consequently of the FAs. The reproducibility of the one-step method was assessed by the measurement of the FAMEs from rapeseed and rapeseed oil. For the C- and H-CSIA of individual FAMEs, a gas chromatography combustion/pyrolysis isotope ratio mass spectrometry system was used. Our results showed that no significant differences arise in the carbon and hydrogen isotope compositions of the selected main FAMEs produced with and without DMP except for the H-CSIA value of C18:3. The reproducibility of the one-step method for rapeseed was in the range of ±0.1 mUr to ± 0.3 mUr for C-CSIA and ±1 mUr to ±3 mUr for H-CSIA of the main FAMEs. DMP improves the transesterification efficiency without influencing the accuracy of the C- and H-CSIA of FAMEs. The performance of the one-step method for rapeseed samples for the determination of C- and H-CSIA values of FAMEs is satisfactory. Thus, the applicability of the one-step method for isotopic fingerprint analyses of FAs in oilseeds is reported for the first time.

Determination of carbon isotope ratios for honey samples by means of a liquid chromatography/isotope ratio mass spectrometry system coupled with a post-column pump.

Liquid chromatography/isotope ratio mass spectrometry (LC/IRMS) has been used to authenticate and trace products such as honey, wine, and lemon juice, and compounds such as caffeine and pesticides. However, LC/IRMS has several disadvantages, including the high cost of the CO2 membrane and blocking by solidified sodium persulfate. Here, we developed an improved system for determining carbon isotope ratios using LC/IRMS. The main improvement was the use of a post-column pump. Using the improved system, we determined δ13 C values for glucose with high accuracy and precision (0.1‰ and 0.1‰, respectively; n=3). The glucose, fructose, disaccharide, trisaccharide, and organic acid constituents of honey samples were analyzed using LC/IRMS. The δ13 C values for glucose, fructose, disaccharides, trisaccharides, and organic acids ranged from -27.0 to -24.2‰, -26.8 to -24.0‰, -28.8 to -24.0‰, -27.8 to -22.8‰, and-30.6 to -27.4‰, respectively. The analysis time was a third to a half of that required for analysis by previously reported methods. The column flow rate could be arbitrarily adjusted with the post-column pump. We applied the improved method to 26 commercial honey samples. Our results can be expected to be useful for other researchers who use LC/IRMS.

Position-specific 13 C/12 C analysis of amino acid carboxyl groups - automated flow-injection analysis based on reaction with ninhydrin.

RationaleThe fundamental level of stable isotopic knowledge lies at specific atomic positions within molecules but existing methods of analysis require lengthy off‐line preparation to reveal this information. An automated position‐specific isotope analysis (PSIA) method is presented to determine the stable carbon isotopic compositions of the carboxyl groups of amino acids (δ 13CCARBOXYL values). This automation makes PSIA measurements easier and routine.MethodsAn existing high‐performance liquid chromatography (HPLC) gas handling interface/stable isotope ratio mass spectrometry system was modified by the addition of a post‐column derivatisation unit between the HPLC system and the interface. The post‐column reaction was optimised to yield CO2 from the carboxyl groups of amino acids by reaction with ninhydrin.ResultsThe methodology described produced δ 13CCARBOXYL values with typical standard deviations below ±0.1 ‰ and consistent differences (Δ 13CCARBOXYL values) between amino acids over a 1‐year period. First estimates are presented for the δ 13CCARBOXYL values of a number of internationally available amino acid reference materials.ConclusionsThe PSIA methodology described provides a further dimension to the stable isotopic characterisation of amino acids at a more detailed level than the bulk or averaged whole‐molecule level. When combined with on‐line chromatographic separation or off‐line fraction collection of protein hydrolysates the technique will offer an automated and routine way to study position‐specific carboxyl carbon isotope information for amino acids, enabling more refined isotopic studies of carbon uptake and metabolism.

Performance of the wet oxidation unit of the HPLC isotope ratio mass spectrometry system for halogenated compounds.

The performance of liquid chromatography-isotope ratio mass spectrometry (LC-IRMS) for polar halogenated compounds was evaluated. Oxidation capacity of the system was tested with halogenated acetic acids and halogenated aromatic compounds. Acetic acid (AA) was selected as a reference compound for complete oxidation and compared on the molar basis to the oxidation of other analytes. The isotope values were proofed with calibrated δ(13)C values obtained with an elemental analyzer (EA). Correct isotope values were obtained for mono- and dichlorinated, fluorinated, and tribrominated acetic acids and also for aniline, phenol, benzene, bromobenzene, chlorobenzene, 1,2-dichlorobenzene, 2,4,6-trichlorophenol, pentafluorophenol, and nitrobenzene. Incomplete oxidation of trichloroacetic acid (TCA) and trifluoroacetic acid (TFA) resulted in lower recovery compared to AA (37% and 24%, respectively) and in isotopic shift compared to values obtained with EA (TCA Δδ(13)C(EA/LC-IRMS) = 8.8‰, TFA Δδ(13)C(EA/LC-IRMS) = 6.0‰). Improvement of oxidation by longer reaction time in the reactor and increase in the concentration of sulfate radicals did not lead to complete combustion of TCA and TFA needed for δ(13)C analysis. To the best of our knowledge, this is the first time such highly chlorinated compounds were studied with the LC-IRMS system. This work provides information for method development of LC-IRMS methods for halogenated contaminants that are known as potential threats to public health and the environment.

Online oxygen isotope analysis of sub‐milligram quantities of biogenic opal using the inductive high‐temperature carbon reduction method coupled with continuous‐flow isotope ratio mass spectrometry

ABSTRACTWe present a method for determining sub‐milligram quantities of biogenic opal. The method employs the inductive high‐temperature carbon reduction method for dehydration of opal and reduction of silica, and a continuous‐flow isotope ratio mass spectrometry system for direct analysis of the oxygen isotope ratio in the evolved carbon monoxide. The accuracy and precision of the online analytical system were evaluated by isotopic analysis of various quantities of standard SiO2 in the range 40–538 µg. The time required to analyse a single sample was relatively short (50 min); hence, our method is suitable for routine analysis for paleoenvironmental studies that require large amounts of time‐series data. The method was validated for samples in the sub‐milligram range and can be applied to oxygen isotope analysis of various types of biogenic opal that have not been analysed because of their small amounts in natural samples. We successfully applied our method to (i) the first oxygen isotope analysis of monospecific radiolarian skeletons and (ii) high‐resolution oxygen isotope analysis of Holocene diatom frustules from the Southern Ocean.

Otolith δ18O of Pacific bluefin tuna Thunnus orientalis as an indicator of ambient water temperature

To detect the relationship between ambient temperature and otolith stable oxygen isotope (δ18O), Pacific bluefin tuna Thunnus orientalis larvae were reared at 6 different temperatures (between 23 and 28°C at 35 psu; mean δ18O value, +0.31‰) for 2 to 8 d after hatching. For the first time, otolith δ18O was measured using a continuous-flow isotope ratio mass spectrometry system. The linear relationship between otolith δ18O and temperature was determined as follows: δotolith – δwater = –2.70 (T °C) + 5.193 (r2 = 0.806, p < 0.01). The temperature-dependent fractionation of otolith δ18O was close to that reported for inorganic aragonite, indicating that the vital effects of isotopes are small. The otolith δ18O values of juveniles reared at a mean of 26.5°C were not significantly different from those of larvae reared at 26°C. With regard to the 2 pairs of otoliths within an individual fish, the differences in δ18O values between the left and right otoliths were subtle. The otolith δ18O values of larvae reared in δ18O-depleted seawater (24 to 26°C at 32 psu; δ18O, −0.17‰) were lower, but were not significantly different from those of larvae reared in +0.31‰ water, which is the δ18O value of the water in which the spawners were raised. These results suggest that the sample size for the 32 psu experiment was too small and that the rearing durations were too short to affect the otoliths completely. Our results demonstrate that otolith δ18O values of Pacific bluefin tuna larvae can provide precise and accurate estimates of their ambient temperature experience.

Long‐term performance of the Gasbench isotope ratio mass spectrometry system for the stable isotope analysis of carbonate microsamples

Long‐term performance of the Gasbench isotope ratio mass spectrometry system for the stable isotope analysis of carbonate microsamples

Offline oxygen isotope analysis of organic compounds with high N:O

Although the advantages of online δ(18)O analysis of organic compounds make its broad application desirable, researchers have encountered NO(+) isobaric interference with CO(+) at m/z 30 (e.g. (14)N(16)O(+), (12)C(18)O(+)) when analyzing nitrogenous substrates. If the δ(18)O value of inter-laboratory standards for substrates with high N:O value could be confirmed offline, these materials could be analyzed periodically and used to evaluate δ(18)O data produced online for nitrogenous unknowns. To this end, we present an offline method based on modifications of the methods of Schimmelmann and Deniro (Anal. Chem. 1985; 57: 2644) and Sauer and Sternberg (Anal. Chem. 1994; 66: 2409), whereby all the N(2) from the gas products of a chlorinated pyrolysis was eliminated, resulting in purified CO(2) for analysis via a dual-inlet isotope ratio mass spectrometry system. We evaluated our method by comparing observed δ(18)O values with previously published or inter-laboratory calibrated δ(18)O values for five nitrogen-free working reference materials; finding isotopic agreement to within ±0.2‰ for SIGMA® cellulose, IAEA-CH3 cellulose (C(6)H(10)O(5)) and IAEA-CH6 sucrose (C(12)H(22)O(11)), and within ±1.8‰ for IAEA-601 and IAEA-602 benzoic acids (C(7)H(6)O(2)). We also compared the δ(18)O values of IAEA-CH3 cellulose and IAEA-CH6 sucrose that was nitrogen-'doped' with adenine (C(5)H(5)N(5)), imidazole (C(3)H(4)N(2)) and 2-aminopyrimidine (C(4)H(5)N(3)) with the undoped δ(18)O values for the same substrates; yielding isotopic agreement to within ±0.7‰. Finally, we provide an independent analysis of the δ(18)O value of IAEA-600 caffeine (C(8)H(10)N(4)O(2)), previously characterized using online systems exclusively, and discuss the reasons for an average 1.4‰ enrichment in δ(18)O observed offline relative to the consensus online δ(18)O value.

A gas chromatography/pyrolysis/isotope ratio mass spectrometry system for high‐precision δD measurements of atmospheric methane extracted from ice cores

Air enclosures in polar ice cores represent the only direct paleoatmospheric archive. Analysis of the entrapped air provides clues to the climate system of the past in decadal to centennial resolution. A wealth of information has been gained from measurements of concentrations of greenhouse gases; however, little is known about their isotopic composition. In particular, stable isotopologues (deltaD and delta(13)C) of methane (CH(4)) record valuable information on its global cycle as the different sources exhibit distinct carbon and hydrogen isotopic composition. However, CH(4) isotope analysis is limited by the large sample size required and the demanding analysis as high precision is required. Here we present a highly automated, high-precision online gas chromatography/pyrolysis/isotope ratio monitoring mass spectrometry (GC/P/irmMS) technique for the analysis of deltaD(CH(4)). It includes gas extraction from ice, preconcentration, gas chromatographic separation and pyrolysis of CH(4) from roughly 500 g of ice with CH(4) concentrations as low as 350 ppbv. Ice samples with approximately 40 mL air and only approximately 1 nmol CH(4) can be measured with a precision of 3.4 per thousand. The precision for 65 mL air samples with recent atmospheric concentration is 1.5 per thousand. The CH(4) concentration can be obtained along with isotope data which is crucial for reporting ice core data on matched time scales and enables us to detect flaws in the measurement procedure. Custom-made script-based processing of MS raw and peak data enhance the system's performance with respect to stability, peak size dependency, hence precision and accuracy and last but not least time requirement.

A gas chromatography/combustion/isotope ratio mass spectrometry system for high‐precision δ13C measurements of atmospheric methane extracted from ice core samples

Past atmospheric composition can be reconstructed by the analysis of air enclosures in polar ice cores which archive ancient air in decadal to centennial resolution. Due to the different carbon isotopic signatures of different methane sources high-precision measurements of delta13CH4 in ice cores provide clues about the global methane cycle in the past. We developed a highly automated (continuous-flow) gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS) technique for ice core samples of approximately 200 g. The methane is melt-extracted using a purge-and-trap method, then separated from the main air constituents, combusted and measured as CO2 by a conventional isotope ratio mass spectrometer. One CO2 working standard, one CH4 and two air reference gases are used to identify potential sources of isotope fractionation within the entire sample preparation process and to enhance the stability, reproducibility and accuracy of the measurement. After correction for gravitational fractionation, pre-industrial air samples from Greenland ice (1831 +/- 40 years) show a delta13C(VPDB) of -49.54 +/- 0.13 per thousand and Antarctic samples (1530 +/- 25 years) show a delta13C(VPDB) of -48.00 +/- 0.12 per thousand in good agreement with published data.

A new continuous flow isotope ratio mass spectrometry system for the analysis of delta2H, delta17O and delta18O of small (120 microg) water samples in atmospheric applications.

A new continuous-flow system for the analysis of the complete stable isotopic composition of water vapor has been developed. The sample size is reduced to only 120 microg (identical with 120 nL of liquid substance) of water, yielding precisions of about 0.7, 1.3 and 7 per thousand for delta17O, delta18O and delta2H, respectively. The total time for the analysis of a sample is about 150 min including purging times. Oxidized steel surfaces can be a source of memory effects which can be corrected for. The system is predestined for atmospheric applications in the tropopause region, as the sample can be directly introduced into the system from a cryogenic trap.

High-precision position-specific isotope analysis of 13C/12C in leucine and methionine analogues.

We report an automated method for high-precision position-specific isotope analysis (PSIA) of carbon in amino acid analogues. Carbon isotope ratios are measured for gas-phase pyrolysis fragments from multiple sources of 3-methylthiopropylamine (3MTP) and isoamylamine (IAA), the decarboxylated analogues of methionine and leucine, using a home-built gas chromatography (GC)-pyrolysis-GC preparation system coupled to a combustion-isotope ratio mass spectrometry system. Over a temperature range of 620-900 degrees C, the characteristic pyrolysis products for 3MTP were CH4, C2H6, HCN, and CH3CN and for IAA products were propylene, isobutylene, HCN, and CH3CN. Fragment origin was confirmed by 13C-labeling, and fragments used for isotope analysis were generated from unique moieties with > 95% structural fidelity. Isotope ratios for the fragments were determined with an average precision of SD(delta13C) < 0.3% per thousand, and relative isotope ratios of fragments from different sources were determined with an average precision of SD(delta(delta)13C) < 0.5% per thousand. Delta(delta)13C values of fragments were invariant over a range of pyrolysis temperatures. The delta(delta)13C of complementary fragments in IAA was within 0.8% per thousand of the delta(delta)13C of the parent compounds, indicating that pyrolysis-induced isotopic fractionation is effectively taken into account with this calibration procedure. Using delta(delta)13C values of fragments, delta(delta)13C values were determined for all four carbon positions of 3MTP and for C1, C2, and the propyl moiety of IAA, either directly or indirectly by mass balance. Large variations in position-specific isotope ratios were observed in samples from different commercial sources. Most dramatically, two 3MTP sources differed by 16.30% per thousand at C1, 48.33% per thousand at C2, 0.37% per thousand at C3, and 5.36% per thousand at C(methyl). These PSIA techniques are suitable for studying subtle changes in intramolecular isotope ratios due to natural processes.

Amount-dependent isotopic fractionation during compound-specific isotope analysis.

The performance of a gas chromatography-combustion-isotope ratio mass spectrometry system (GC-C-IRMS) with respect to the dependence of delta(13)C values on the amount of sample is presented. Particular attention is paid to the localization of the amount-dependent isotopic fractionation within the system. Injection experiments with varying amounts of gases (CO(2), n-hexane, and toluene) revealed that neither the detector unit nor the combustion reactor, but rather the conditions in the split/splitless injector, contributed to this effect. Although optimization of injector parameters was performed and a reduction of this adverse effect from 3 to 1 per thousand was achieved, it was not possible to eliminate isotopic fractionation completely. Consequently, additional injector parameters have to be considered and adjusted to achieve injection conditions free of fractionation. For routine analysis of the compound-specific delta(13)C analysis of different biomarkers in many environmental samples, perfect optimization may not always be reached. Therefore, in order to prevent systematic errors in the measured delta(13)C values due to different sample concentrations, it is suggested that correction for the remaining unknown amount-dependent fractionation can be made by means of co-analyzing standards of varying analyte concentrations and known delta(13)C values. Residual overall amount-dependent isotope-fractionation can thus be corrected mathematically.

Comparison of a dual‐inlet gas isotope ratio mass spectrometry system and an automated single‐inlet mass spectrometry system for δ13C analysis

Abstract A dual‐inlet isotope ratio mass spectrometer (VG‐SIRA 12, Isotech, Middlewich, England) using sealed‐glass combustion sample preparation and an automated single‐inlet isotope ratio mass spectrometry system (RoboPrep Sample Converter interfaced with a TracerMass Stable Isotope Detector, Europa Scientific, Crewe, England) were compared for δ13C analysis. Two hundred twenty four samples of plant and animal tissue were analyzed for S13C using the two systems. Delta 13C values obtained from the two instruments were highly correlated (r2=0.986, P=0.0001). The slope of the relationship was not different from one nor was the Y‐intercept different from zero (Y=‐0.2 + 1.0X; P=0.86 and P=0.28 for the slope and Y‐intercept, respectively). In addition, samples from two complete experiments were run on both mass spectrometery systems and statistical analyses performed. In both experiments, differences between treatments were detectable using the RoboPrep system that were not detectable using the SIRA 12 system...