Determination of (13)CO(2)/(12)CO(2) Ratio by IRMS and NDIRS

Abstract Breath tests using (13)C-labelled substrates require the measurement of (13)CO(2)/(12)CO(2) ratio in breath gas samples. Next to isotope ratio mass spectrometry (IRMS), which is very sensitive but also complex and expensive, alternatively isotope selective nondispersive infrared spectrometry (NDIRS) can be used to determine the (13)CO(2)/(12)CO(2) ratio in expired breath. In this study we compared NDIRS- with IRMS-results to investigate whether the less expensive and very simply to operate NDIRS works as reliable as IRMS. For this purpose we applicated 1-(13)C-Phenylalanine to patients with advanced liver cirrhosis and healthy volunteers and took duplicated breath samples for IRMS and NDIRS at selected time points. Our data show a good correlation between these two methods for a small number of samples as required for simple breath tests. Longer series, where repeated measurements are required on the NDIRS instrument lead to a decreasing correlation. This indicates the superiority of IRMS concerning (13)CO(2)-kinetics over longer time periods.     

A Sensitive Isotope Selective Nondispersive Infrared Spectrometer for 13CO2 and 12CO2 Concentration Measurements in Breath Samples

We present a nondispersive infrared spectrometer (NDIRS) for the measurement of the ¹³CO₂/¹²CO₂-ratio in breath samples. A commercial NDIR spectrometer for CO₂ concentration measurements in industrial process control was modified using two separate optical channels for the ¹³CO₂ and ¹²CO₂ detection. Cross interference due to overlapping absorption lines of both isotopic gases was successfully eliminated. The sensitivity of this device is ± 0.4‰ of the ¹³CO₂/¹²CO₂-ratio in a range of 2.5 to 5% of total CO₂. This is sufficient for biomedical applications. Our spectrometer is small in size, cheap and simple to operate and thus a true alternative to isotope ratio mass spectrometers (IRMS). Several biomedical applications with breath samples were demonstrated and were compared in very good agreement with IRMS.     

Biomedical Application of an Isotope Selective Nondispersive Infrared Spectrometer for 13CO2 and 12CO2 Concentration Measurements in Breath Samples

A newly developed isotope selective nondispersive infrared (NDIR) spectrometer for the measurement of ¹³CO₂ and ¹²CO₂ concentrations in breath samples was applied as a low cost and very simple to operate alternative to isotope ratio mass spectrometry (IRMS). We used this device for several biomedical applications ([¹³C]urea breath test, [¹³C]leucine metabolism, [¹³C]methacetin catabolism of rats) and found that the results agree very well with IRMS.     

Determination of 13C/12C-Ratios in Rumen Produced Methane and CO2 of Cows,Sheep and Camels

Abstract

Naturally produced methane shows different δ¹³C-values with respect to its origin, e.g., geological or biological. Methane-production of ruminants is considered to be the dominant source from the animal kingdom. Isotopic values of rumen methane—given in literature—range between ?80‰ and -50‰ and are related to feed composition and also sampling techniques. Keeping cows, camels and sheep under identical feed conditions and sampling rumen gases via implanted fistulae we compared δ_PDB ¹³C-values of methane and CO₂ between the species. Referring to mean values obtained from 4 or 5 samples at different times of 11 animals (n = 47) we calculated δ_PDB ¹³C-medians resulting in small but not significant differences within and significant differences between the species for CO₂ and methane. The δ_PDB ¹³C-differences between methane and CO₂ were statistically equal within and also between the species. Therefore a linear regression of methane values on CO₂ is appropriate and leads to: δ_PDB ¹³C(methane)‰ = 1,57 * δ_PDB ¹³C(CO₂)‰-47‰ with a correlation coefficient of r = 0,87.     

CO2及其碳同位素比值高精度检测研究

改进了一种基于傅里叶变换红外光谱法测量CO₂气体的装置, 改进后的装置能够提高CO₂检测精度, 并能同时测量CO₂碳同位素比值. 研究了温度和压力对CO₂浓度值和CO₂碳同位素比值测量的影响规律. 利用该装置连续测量了标准CO₂气体和环境大气, 对标准CO₂气体测量得到的CO₂浓度值及其碳同位素比值进行温度和压力影响修正, 获得了较好的精度和准确度. 关键词： 光谱学 同位素比值 傅里叶变换红外光谱 二氧化碳     

Carbon Isotope Analysis in Urea at High 13C-Abundances Using the 13/12CO2-Breath Test Device Fanci2

Abstract

The increasing application of ¹³C-labelled urea in medicine requires simple and reasonable methods for measuring highly enriched ¹³C in urea. The combination: ultimate organic analysis—mass spectrometry so far prescribed is complicated and expensive. For medical diagnosis, however, isotope selective nondispersive infrared spectrometers (NDIRS) have been available for many years. One of these tools is FANci2 which is very reasonable and easily to be operated. By means of such devices also urea highly enriched in ¹³C can be analysed, provided that the samples are first diluted with a defined amount of urea of natural isotopic composition and then transformed into carbon dioxide by means of urease. The relative abundance of ¹³C in this carbon dioxide, measured by nondispersive infrared spectrometry, is then a measure of the ¹³C abundance in the initial urea sample. Comparison of results of such measurements with those attained by mass spectrometry proves that this procedure is feasible and yields precise results.     

Macroscopic Isotope Separation of 13C by a CO2 Laser

By photochemical dissociation of the rare carbon isotope component of CHClF₂ by means of a CO₂ laser with an average power of 150 W, Q-switched at 10 kHz, we have demonstrated the separation of more than 1 mol of ¹³C, enriched to 50% (2 mol of total carbon). It is contained in about 1 mol (101 g) of the product C₂F₄. The total throughput of the starting material was 29 kg. The experiment was run day and night for 2 weeks, almost only controlled by a computer. We obtained production rates of 5 mmol/h, corresponding to about 0.5 kg ¹³C per year.     

Microcombustion of ng Amounts of Carbon in Non-Volatile Materials for isotope Ratio Evaluation

Abstract

A novel microcombustion technique for carbon isotopic analysis of nanogram amounts of carbon in non-volatile materials based on isotope ratio monitoring (irm) mass spectrometry is described. Liquid or solid samples placed in a quartz sleeve are combusted at 1000°C in a continuous stream of helium and oxygen. CO₂ removed from the carrier gas stream by cryogenic trapping is transferred onto a GC column. Following GC separation, the CO₂ is transferred via an open split to the ion source of a gas isotope ratio mass spectrometer. Reproducibility for samples >25 nmol carbon is <1‰. Problems associated with blanks from various sources and with reproducible deposition of small sample amounts led to variable accuracy, which was dependent on the compound class being analysed. Minimum sample size was in the range from 5 to 10 nmol carbon. Measurements of dissolved organic carbon (DOC) of groundwater from Germany yielded consistent values of δ¹³C = -28.8‰.     

Analytics/Methods

An overview is presented of biomedical applications of stable isotopes in general, but mainly focused on the activities of the Center for Liver, Digestive and Metabolic Diseases of the University Medical Center Groningen. The aims of metabolic studies in the areas of glucose, fat, cholesterol and protein metabolism are briefly explained, as well as the principle of breath testing and the techniques to study body composition and energy expenditure. Much attention is paid to the analytical considerations based upon metabolite concentrations, sample size restrictions, the availability of stable isotope labelled substrates and dose requirements in relation to compound-specific isotope analysis. The instrumental advantages and limitations of the generally used techniques gas chromatography/reaction/isotope ratio mass spectrometry and gas chromatography/mass spectrometry are described as well as the novelties of the recently commercialised liquid chromatography/combustion/isotope ratio mass spectrometry. The present use and future perspective of infrared (IR) spectrometry for clinical and biomedical stable isotope applications are reviewed. In this respect, the analytical demands on IR spectrometry are discussed to enable replacement of isotope ratio mass spectrometry by IR spectrometry, in particular, for the purpose of compound-specific isotope ratio analysis in biological matrices.     

Use of the optogalvanic effect (OGE) for isotope ratio spectrometry of 13CO2 and 14CO2

The use of isotopic carbon dioxide lasers for determination of carbon (and oxygen) isotope ratios was first demonstrated in 1994. Since then a commercial device called LARA^?, has been manufactured and used for Helicobacter pylori breath tests using ¹³C-labelled urea. The major advantages of the optogalvanic effect compared with other infrared absorption isotope ratio measurement techniques are its lack of optical background and its high sensitivity resulting from a signal gain proportional to laser power. Continuous normalisation using two cells, a standard and sample, lead to high accuracy as well as precision. Recent advances in continuous flow measurement of ¹³C/¹²C ratios of CO₂ in air and extensions of the technique to ¹⁴C, which can be analysed as a stable isotope, are described.     

13C Basal Abundance of Expired CO2 -Definition of Pre-Requisites for Kinetic Breath Tests

Abstract

A sufficiently stable rate of ¹³CO₂ exhalation is necessary when the diagnostic ¹³CO₂ breath tests are performed in healthy subjects and patients. The aim of the research was to define prerequisite conditions for kinetic breath tests in order to ensure a stable ¹³CO₂ background. A 3-part protocol was developed. Part I: a study of the one-day variation of ¹³CO₂ abundance in expired CO₂ confirmed that shifts of the basal ¹³C abundance in breath are inherent in nature. Part II: a study of the variations of ¹³C enrichment after the ingestion of different meals and beverages showed that ingestion of food items containing C₄ plant sugars, such as maize, induces a significant increase in isotopic abundance. Part III: a new test breakfast containing rice grain cereal, milk and orange juice was tested. This test meal induces no significant change on the basal ¹³CO₂ abundance in healthy subjects. This new finding allows to avoid the fasting period normally required prior to a breath test which is sometimes difficult for children and pregnant women.     

二元比例X射线荧光光谱法测定BaFe12O19中Fe和Ba的含量

采用二元比例X射线荧光光谱法测定铁钡氧体BaFe12O19中铁(Fe)和钡(Ba)的含量,为铁钡氧体BaFe12O19的定量分析建立了一种新的分析方法.选取纯Fe粉(分析纯)及纯BaCO3粉(分析纯),采用粉末压片法自制标准样品,建立了定标曲线.选用Fe含量为75％、Ba含量为17.4％的标准样品作为验证样品,对定标曲线的准确度进行验证.经分析得到验证样品中Fe和Ba含量分别为75.68％和16.92％,相对标准偏差分别为0.317％和1.11％,可知定标曲线具有良好的线性关系,且准确度高,精密度好.实验测得BaFe12O19样品中Fe和Ba的含量分别为57.28％和13.42％,相对标准偏差为1.05％和0.73％.     

基于傅里叶变换红外光谱法CO2气体碳同位素比检测研究

介绍了基于傅里叶变换红外技术检测CO₂气体碳同位素比的新方法, 详细介绍了如何从HITRAN红外数据库中提取气体标准吸收截面; 介绍了基于非线性最小二乘法反演CO₂气体碳同位素比和整套实验装置的组成及实验步骤. 从理论和实验分析两方面讨论了温度和气压变化对δ¹³CO₂值的影响规律. 对于同一CO₂标准气体, 采用FTIR和同位素质谱法两种技术进行了δ¹³CO₂值对比检测, 两种测量技术的平均值差异仅为0.25%. 从实验结果可以看出, FTIR技术可以实现对CO₂气体碳同位素比的检测.     

用HI-13串列加速器质谱装置测量182Hf的研究

¹⁸²Hf的半衰期为(8.90±0.09)Ma, 是一个接近灭绝的放射性核素. 超新星爆炸是自然界中已知的惟一能产生¹⁸²Hf的途径. 因此¹⁸²Hf是研究近1亿年来在地球附近可能发生的超新星事件的理想核素. 另外, ¹⁸²Hf是核工程中特别感兴趣的一个长寿命放射性核素. 精确测量超痕量的¹⁸²Hf对反应堆的设计和核天体物理学以及其他研究领域都是非常重要的. 用加速器质谱有可能实现对超低含量¹⁸²Hf的测量. 在中国原子能科学研究院的HI-13加速器质谱装置上对¹⁸²Hf的测量方法以及样品的化学去钨方法进行了研究, 分别得到了空白样品以及系列标准样品的¹⁸²Hf和¹⁸³W的能量-飞行时间双维谱. ¹⁸²W对¹⁸²Hf计数的贡献是通过测量¹⁸³W的计数归一扣除的. 目前本工作对¹⁸²Hf的测量灵敏度为4.15±10^－11 (¹⁸²Hf/¹⁸⁰Hf比值).     

13C qNMR结合DFSS方法测定油品芳碳率

利用六西格玛设计（design for six Sigma,DFSS）方法,通过定量核磁共振碳谱（quantitative carbon nuclearmagnetic resonance spectroscopy,¹³C qNMR）对由11种已知烃类化合物组成的模型油品进行了芳香碳摩尔百分比C（ar）%,简称芳碳率]测定的系统研究,建立了C（ar）%与样品浓度Conc.、弛豫试剂浓度[Cr（acac）₃]和循环延迟D13个参数之间的数学模型,据此模型得到了优化的参数设置：Conc.为180 mg/mL,[Cr（acac）₃]为12 mg/mL,D1为10.5 s.按此设置进行了8组验证实验,其结果重现性较好,相对标准偏差（relative standard deviation,RSD）为0.61%;准确性较高,达到99.0%~100.6%.该方法也可用于其它分析测试方法的系统研究,且实验结果对现行油品芳碳率测定标准中相关参数,如[Cr（acac）₃]和D1的设置能够提出补充性建议.     

The13C/12C and 2H/1H Ratios of Trichloroethene,Tetrachloroethene and Their Metabolites

Abstract

Isotope ratios of technically and biologically produced halogenated hydrocarbons (HHC) are presented. Compared to the parent compounds, there is a significant enrichment of deuterium depending on the manner of production, whereas the carbon isotope ratio was only slightly influenced. Furthermore, there is nearly no alteration of the carbon isotope ratio due to biological degradation, so that the carbon fingerprint of the parent compounds is maintained in the metabolites. In contrast to this, the hydrogen isotope ratio of the metabolites reflects the ratio of the surrounding water. These results show that the isotopic fingerprint is a useful tool to distinguish between age, different charges of chemical dumping, and in addition to that, whether there is a contamination with technical products or residual metabolites.     

The effect of physical back-diffusion of 13CO2 tracer on the coupling between photosynthesis and soil CO2 efflux in grassland

Pulse labelling experiments provide a common tool to study short-term processes in the plant–soil system and investigate below-ground carbon allocation as well as the coupling of soil CO₂ efflux to photosynthesis. During the first hours after pulse labelling, the measured isotopic signal of soil CO₂ efflux is a combination of both physical tracer diffusion into and out of the soil as well as biological tracer release via root and microbial respiration. Neglecting physical back-diffusion can lead to misinterpretation regarding time lags between photosynthesis and soil CO₂ efflux in grassland or any ecosystem type where the above-ground plant parts cannot be labelled in gas-tight chambers separated from the soil. We studied the effects of physical ¹³CO₂ tracer back-diffusion in pulse labelling experiments in grassland, focusing on the isotopic signature of soil CO₂ efflux. Having accounted for back-diffusion, the estimated time lag for first tracer appearance in soil CO₂ efflux changed from 0 to 1.81±0.56 h (mean±SD) and the time lag for maximum tracer appearance from 2.67±0.39 to 9.63±3.32 h (mean±SD). Thus, time lags were considerably longer when physical tracer diffusion was considered. Using these time lags after accounting for physical back-diffusion, high nocturnal soil CO₂ efflux rates could be related to daytime rates of gross primary productivity (R²=0.84). Moreover, pronounced diurnal patterns in the δ¹³C of soil CO₂ efflux were found during the decline of the tracer over 3 weeks. Possible mechanisms include diurnal changes in the relative contributions of autotrophic and heterotrophic soil respiration as well as their respective δ¹³C values. Thus, after accounting for physical back-diffusion, we were able to quantify biological time lags in the coupling of photosynthesis and soil CO₂ efflux in grassland at the diurnal time scale.     

Determination of Isotope Enrichment in 13C-or 2H-Labelled Branched-Chain l-Amino Acids From Physiological Fluids by Gas Chromatography-Mass Spectrometry: Use of l-Leucine Dehydrogenase for Specific Preparation of the Quinoxalinol Derivatives

A procedure is described which allows the specific and sensitive estimation of ¹³C- or ²H-label enrichment in branched-chain l-amino acids from physiological fluids: Amino acids are isolated from deproteinized samples by cation exchange chromatography. Specific conversion of branched-chain l-amino acids to their respective 2-oxo acids is achieved by treatment with l-leucine dehydrogenase from Bacillus sp. Reaction of the 2-oxo acids with o-phenylenediamin yields the branched-chain quinoxalinol derivatives which are purified by solid phase extraction. Isotopic label enrichment is then determined by gas chromatographic-mass spectrometric analysis of the O-trimethylsilyl quinoxalinol derivatives using an ammonia-chemical ionisation mode and selected ion monitoring of the quasi-molecular ions [MH]⁺ and [MH + 1]⁺. Applicability of the method is demonstrated by serum analysis in an oral loading test with l-[1-¹³C]leucine.     

Evaluation of Water Removal and Memory Effect in 13CO2 Breath Tests by Isotope Ratio Mass Spectrometry

The usefulness of different ways of water removal in off-line sample preparation of human breath samples for ¹³CO₂ breath tests was examined and compared. Cryogenic water trapping and water removal with common desiccants like silicagel blue, Mg(ClO₄)₂, and molecular sieves were checked for reliability and reproducibility. With silicagel blue and Mg(ClO₄)₂ memory effects for ¹³C content were observed. The use of molecular sieve 4 Å and 5 Å led to tremendous carbon isotope fractionation. Molecular sieve 3 Å was found to be an excellent alternative to the established use of Mg(ClO₄)₂ and of cryogenic water trapping.     

双波长比值光谱法测定复方乙酰水杨酸片中三组分含量

应用双波长比值光谱法测定了复方乙酰水杨酸片中三组分的含量。依据复方乙酰水杨酸片中三组分的比值光谱特征, 选择213,227,245和265 nm作为测定三个组分的波长。结果显示乙酰水杨酸在5～20 μg·mL-1, 非那西丁在2～10 μg·mL-1, 咖啡因在2～20 μg·mL-1浓度范围内线性关系良好,平均回收率分别为100.03%,100.23%和99.96%。样品测定结果与部颁标准方法一致(P＞0.05)。本法具有测定波长少、计算简单、光谱“分离”能力强以及能在低档分光光度计上实现, 易于推广等特点。