Detection by mass spectrometry
A mass spectrum of an organic compound will usually contain a small peak of one mass unit greater than the apparent molecular ion peak (M) of the whole molecule. This is known as the M+1 peak and comes from the few molecules that contain a 13C atom in place of a 12C. A molecule containing one carbon atom will be expected to have an M+1 peak of approximately 1.1% of the size of the M peak, as 1.1% of the molecules will have a 13C rather than a 12C. Similarly, a molecule containing two carbon atoms will be expected to have an M+1 peak of approximately 2.2% of the size of the M peak, as there is double the previous likelihood that any molecule will contain a 13C atom.
In the above, the mathematics and chemistry have been simplified, however it can be used effectively to give the number of carbon atoms for small- to medium-sized organic molecules. In the following formula the result should be rounded to the nearest integer:
where C = number of C atoms, X = amplitude of the M ion peak, and Y = amplitude of the M +1 ion peak.
13C-enriched compounds are used in the research of metabolic processes by means of mass spectrometry. Such compounds are safe because they are non-radioactive. In addition, 13C is used to quantify proteins (quantitative proteomics). One important application is in stable isotope labeling by amino acids in cell culture (SILAC). 13C-enriched compounds are used in medical diagnostic tests such as the urea breath test. Analysis in these tests is usually of the ratio of 13C to 12C by isotope ratio mass spectrometry.
The ratio of 13C to 12C is slightly higher in plants employing C4 carbon fixation than in plants employing C3 carbon fixation. Because the different isotope ratios for the two kinds of plants propagate through the food chain, it is possible to determine if the principal diet of a human or other animal consists primarily of C3 plants or C4 plants by measuring the isotopic signature of their collagen and other tissues.
Uses in science
Due to differential uptake in plants as well as marine carbonates of 13C, it is possible to use these isotopic signatures in earth science. Biological processes preferentially take up the lower mass isotope through kinetic fractionation. In aqueous geochemistry, by analyzing the δ13C value of carbonaceous material found in surface and ground waters, the source of the water can be identified. This is because atmospheric, carbonate, and plant derived δ13C values all differ. In biology, the ratio of carbon-13 and carbon-12 isotopes in plant tissues is different depending on the type of plant photosynthesis and this can be used, for example, to determine which types of plants were consumed by animals. Greater carbon-13 concentrations indicate stomatal limitations, which can provide information on plant behaviour during drought. [2] Tree ring analysis of carbon isotopes can be used to retrospectively understand forest photosynthesis and how it is impacted by drought. [3]
In geology, the 13C/12C ratio is used to identify the layer in sedimentary rock created at the time of the Permian extinction 252 Mya when the ratio changed abruptly by 1%. More information about usage of 13C/12C ratio in science can be found in the article about isotopic signatures.
Carbon-13 has a non-zero spin quantum number of ½, and hence allows the structure of carbon-containing substances to be investigated using carbon-13 nuclear magnetic resonance.
The carbon-13 urea breath test is a safe and highly accurate diagnostic tool to detect the presence of Helicobacter pylori infection in the stomach. [4] The urea breath test utilizing carbon-13 is preferred to carbon-14 for certain vulnerable populations due to its non-radioactive nature. [4]
The urea breath test is a rapid diagnostic procedure used to identify infections by Helicobacter pylori, a spiral bacterium implicated in gastritis, gastric ulcer, and peptic ulcer disease. It is based upon the ability of H. pylori to convert urea to ammonia and carbon dioxide. Urea breath tests are recommended in leading society guidelines as a preferred non-invasive choice for detecting H. pylori before and after treatment.
Carbon-14, C-14, 14
C or radiocarbon, is a radioactive isotope of carbon with an atomic nucleus containing 6 protons and 8 neutrons. Its presence in organic materials is the basis of the radiocarbon dating method pioneered by Willard Libby and colleagues (1949) to date archaeological, geological and hydrogeological samples. Carbon-14 was discovered on February 27, 1940, by Martin Kamen and Sam Ruben at the University of California Radiation Laboratory in Berkeley, California. Its existence had been suggested by Franz Kurie in 1934.
Ureases, functionally, belong to the superfamily of amidohydrolases and phosphotriesterases. Ureases are found in numerous bacteria, fungi, algae, plants, and some invertebrates, as well as in soils, as a soil enzyme. They are nickel-containing metalloenzymes of high molecular weight.
Isotope analysis is the identification of isotopic signature, abundance of certain stable isotopes of chemical elements within organic and inorganic compounds. Isotopic analysis can be used to understand the flow of energy through a food web, to reconstruct past environmental and climatic conditions, to investigate human and animal diets, for food authentification, and a variety of other physical, geological, palaeontological and chemical processes. Stable isotope ratios are measured using mass spectrometry, which separates the different isotopes of an element on the basis of their mass-to-charge ratio.
A radioactive tracer, radiotracer, or radioactive label is a synthetic derivative of a natural compound in which one or more atoms have been replaced by a radionuclide. By virtue of its radioactive decay, it can be used to explore the mechanism of chemical reactions by tracing the path that the radioisotope follows from reactants to products. Radiolabeling or radiotracing is thus the radioactive form of isotopic labeling. In biological contexts, experiments that use radioisotope tracers are sometimes called radioisotope feeding experiments.
Isotope geochemistry is an aspect of geology based upon the study of natural variations in the relative abundances of isotopes of various elements. Variations in isotopic abundance are measured by isotope-ratio mass spectrometry, and can reveal information about the ages and origins of rock, air or water bodies, or processes of mixing between them.
Isotopic labeling is a technique used to track the passage of an isotope through chemical reaction, metabolic pathway, or a biological cell. The reactant is 'labeled' by replacing one or more specific atoms with their isotopes. The reactant is then allowed to undergo the reaction. The position of the isotopes in the products is measured to determine the sequence the isotopic atom followed in the reaction or the cell's metabolic pathway. The nuclides used in isotopic labeling may be stable nuclides or radionuclides. In the latter case, the labeling is called radiolabeling.
Isotopomers or isotopic isomers are isomers which differ by isotopic substitution, and which have the same number of atoms of each isotope but in a different arrangement. For example, CH3OD and CH2DOH are two isotopomers of monodeuterated methanol.
Carbon (6C) has 14 known isotopes, from 8
C
to 20
C
as well as 22
C
, of which 12
C
and 13
C
are stable. The longest-lived radioisotope is 14
C
, with a half-life of 5.70(3)×103 years. This is also the only carbon radioisotope found in nature, as trace quantities are formed cosmogenically by the reaction 14
N
+
n
→ 14
C
+ 1
H
. The most stable artificial radioisotope is 11
C
, which has a half-life of 20.3402(53) min. All other radioisotopes have half-lives under 20 seconds, most less than 200 milliseconds. The least stable isotope is 8
C
, with a half-life of 3.5(1.4)×10−21 s. Light isotopes tend to decay into isotopes of boron and heavy ones tend to decay into isotopes of nitrogen.
An isotopic signature is a ratio of non-radiogenic 'stable isotopes', stable radiogenic isotopes, or unstable radioactive isotopes of particular elements in an investigated material. The ratios of isotopes in a sample material are measured by isotope-ratio mass spectrometry against an isotopic reference material. This process is called isotope analysis.
Carbon-13 (C13) nuclear magnetic resonance is the application of nuclear magnetic resonance (NMR) spectroscopy to carbon. It is analogous to proton NMR and allows the identification of carbon atoms in an organic molecule just as proton NMR identifies hydrogen atoms. 13C NMR detects only the 13
C
isotope. The main carbon isotope, 12
C
does not produce an NMR signal. Although ca. 1 mln. times less sensitive than 1H NMR spectroscopy, 13C NMR spectroscopy is widely used for characterizing organic and organometallic compounds, primarily because 1H-decoupled 13C-NMR spectra are more simple, have a greater sensitivity to differences in the chemical structure, and, thus, are better suited for identifying molecules in complex mixtures. At the same time, such spectra lack quantitative information about the atomic ratios of different types of carbon nuclei, because nuclear Overhauser effect used in 1H-decoupled 13C-NMR spectroscopy enhances the signals from carbon atoms with a larger number of hydrogen atoms attached to them more than from carbon atoms with a smaller number of H's, and because full relaxation of 13C nuclei is usually not attained, and the nuclei with shorter relaxation times produce more intense signals.
The mass recorded by a mass spectrometer can refer to different physical quantities depending on the characteristics of the instrument and the manner in which the mass spectrum is displayed.
The Suess effect is a change in the ratio of the atmospheric concentrations of heavy isotopes of carbon (13C and 14C) by the admixture of large amounts of fossil-fuel derived CO2, which contains no 14CO2 and is depleted in 13CO2 relative to CO2 in the atmosphere and carbon in the upper ocean and the terrestrial biosphere. It was discovered by and is named for the Austrian chemist Hans Suess, who noted the influence of this effect on the accuracy of radiocarbon dating. More recently, the Suess effect has been used in studies of climate change. The term originally referred only to dilution of atmospheric 14CO2 relative to 12CO2. The concept was later extended to dilution of 13CO2 and to other reservoirs of carbon such as the oceans and soils, again relative to 12C.
In geochemistry, paleoclimatology, and paleoceanography δ13C is an isotopic signature, a measure of the ratio of the two stable isotopes of carbon—13C and 12C—reported in parts per thousand. The measure is also widely used in archaeology for the reconstruction of past diets, particularly to see if marine foods or certain types of plants were consumed.
The start of the Cambrian period is marked by "fluctuations" in a number of geochemical records, including Strontium, Sulfur and Carbon isotopic excursions. While these anomalies are difficult to interpret, a number of possibilities have been put forward. They probably represent changes on a global scale, and as such may help to constrain possible causes of the Cambrian explosion.
Isotopic analysis by nuclear magnetic resonance allows the user to quantify with great precision the differences of isotopic contents on each site of a molecule and thus to measure the specific natural isotope fractionation for each site of this molecule. The SNIF-NMR analytical method was developed to detect the (over) sugaring of wine and enrichment of grape musts, and is mainly used to check the authenticity of foodstuffs and to control the naturality of some aromatic molecules. The SNIF-NMR method has been adopted by the International Organisation of Vine and Wine (OIV) and the European Union as an official method for wine analysis. It is also an official method adopted by the Association Of Analytical Chemists (AOAC) for analysis of fruit juices, maple syrup, vanillin, and by the European Committee for Standardization (CEN) for vinegar.
Position-specific isotope analysis, also called site-specific isotope analysis, is a branch of isotope analysis aimed at determining the isotopic composition of a particular atom position in a molecule. Isotopes are elemental variants with different numbers of neutrons in their nuclei, thereby having different atomic masses. Isotopes are found in varying natural abundances depending on the element; their abundances in specific compounds can vary from random distributions due to environmental conditions that act on the mass variations differently. These differences in abundances are called "fractionations," which are characterized via stable isotope analysis.
Methane clumped isotopes are methane molecules that contain two or more rare isotopes. Methane (CH4) contains two elements, carbon and hydrogen, each of which has two stable isotopes. For carbon, 98.9% are in the form of carbon-12 (12C) and 1.1% are carbon-13 (13C); while for hydrogen, 99.99% are in the form of protium (1H) and 0.01% are deuterium (2H or D). Carbon-13 (13C) and deuterium (2H or D) are rare isotopes in methane molecules. The abundance of the clumped isotopes provides information independent from the traditional carbon or hydrogen isotope composition of methane molecules.
Photosynthesis converts carbon dioxide to carbohydrates via several metabolic pathways that provide energy to an organism and preferentially react with certain stable isotopes of carbon. The selective enrichment of one stable isotope over another creates distinct isotopic fractionations that can be measured and correlated among oxygenic phototrophs. The degree of carbon isotope fractionation is influenced by several factors, including the metabolism, anatomy, growth rate, and environmental conditions of the organism. Understanding these variations in carbon fractionation across species is useful for biogeochemical studies, including the reconstruction of paleoecology, plant evolution, and the characterization of food chains.
The stable isotope composition of amino acids refers to the abundance of heavy and light non-radioactive isotopes of carbon, nitrogen, and other elements within these molecules. Amino acids are the building blocks of proteins. They are synthesized from alpha-keto acid precursors that are in turn intermediates of several different pathways in central metabolism. Carbon skeletons from these diverse sources are further modified before transamination, the addition of an amino group that completes amino acid biosynthesis. Bonds to heavy isotopes are stronger than bonds to light isotopes, making reactions involving heavier isotopes proceed slightly slower in most cases. This phenomenon, known as a kinetic isotope effect, gives rise to isotopic differences between reactants and products that can be detected using isotope ratio mass spectrometry. Amino acids are synthesized via a variety of pathways with reactions containing different, unknown isotope effects. Because of this, the 13C content of amino acid carbon skeletons varies considerably between the amino acids. There is also an isotope effect associated with transamination, which is apparent from the abundance of 15N in some amino acids.