Quantitation isotope dilution

Colby, B. N., Colby, M. E., and Rosecrance, A. E. (1981). Measurement parameter selection for quantitative isotope dilution gas chromatography/mass spectrometry. Anal. Chem. 53, 1907-1911. 

Three common quantitative applications of radiochemical methods of analysis are considered in this section the direct analysis of radioactive isotopes by measuring their rate of disintegration, neutron activation, and the use of radioactive isotopes as tracers in isotope dilution. 

Isotope Dilution Another important quantitative radiochemical method is isotope dilution. In this method of analysis a sample of analyte, called a tracer, is prepared in a radioactive form with a known activity. Ax, for its radioactive decay. A measured mass of the tracer, Wf, is added to a sample containing an unknown mass, w, of a nonradioactive analyte, and the material is homogenized. The sample is then processed to isolate wa grams of purified analyte, containing both radioactive and nonradioactive materials. The activity of the isolated sample, A, is measured. If all the analyte, both radioactive and nonradioactive, is recovered, then A and Ax will be equal. Normally, some of the analyte is lost during isolation and purification. In this case A is less than Ax, and... 

Radiochemical methods of analysis take advantage of the decay of radioactive isotopes. A direct measurement of the rate at which a radioactive isotope decays may be used to determine its concentration in a sample. For analytes that are not naturally radioactive, neutron activation often can be used to induce radioactivity. Isotope dilution, in which a radioactively labeled form of an analyte is spiked into the sample, can be used as an internal standard for quantitative work. 

A major example of isotope-dilution analysis lies in the procedure itself, which does not require any quantitative isolation of the elements being investigated. The relation between the abundance of the element under investigation and the spike is such that, once the spike has been intimately mixed with the sample, any losses of sample have no effect on the result (Figure 48.14). 

ICPMS is uniquely able to borrow a quantitation technique from molecular mass spectrometry. Use of the isotope dilution technique involves the addition of a spike having a different isotope ratio to the sample, which has a known isotope ratio. This is usefiil for determining the concentration of an element in a sample that must undergo some preparation before analysis, or for measuring an element with high precision and accuracy. ... 

Since the carrier effect is not general for all analytes and all additives, quantitative studies using the particle-beam interface should only be carried out after a very careful choice of experimental conditions and standard(s) to be used, with isotopic-dilution methodology being advocated for the most accurate results. 

The particle-beam interface has been developed primarily to provide El spectra from HPLC eluates but may be combined with other ionization techniques such as CL If quantitative studies are being undertaken, a detailed study of experimental conditions should be undertaken. Isotope-dilution methodology is advocated for the most accurate results. 

Dizdaroglu, M. (1993). Quantitative determination of oxidative base damage in DNA by stable isotope-dilution mass spectrometry. FEBS Lett. 315, 1-6. 

Quantitative analysis using FAB is not straightforward, as with all ionisation techniques that use a direct insertion probe. While the goal of the exercise is to determine the bulk concentration of the analyte in the FAB matrix, FAB is instead measuring the concentration of the analyte in the surface of the matrix. The analyte surface concentration is not only a function of bulk analyte concentration, but is also affected by such factors as temperature, pressure, ionic strength, pH, FAB matrix, and sample matrix. With FAB and FTB/LSIMS the sample signal often dies away when the matrix, rather than the sample, is consumed therefore, one cannot be sure that the ion signal obtained represents the entire sample. External standard FAB quantitation methods are of questionable accuracy, and even simple internal standard methods can be trusted only where the analyte is found in a well-controlled sample matrix or is separated from its sample matrix prior to FAB analysis. Therefore, labelled internal standards and isotope dilution methods have become the norm for FAB quantitation. 

The main advantages of plasma-source mass spectrometry (PS-MS) over other analytical techniques, such as PS-AES and ETAAS, are the possibilities of quantitative isotope determination and isotope dilution analysis the rapid spectral scanning capability of the mass spectrometer and semiquantitative determinations to within a factor of two or three. Several labelling methods are used for the quantification of analytes present in complex mixtures. In these methods, the sample is spiked... 

Table 8.62 shows the main characteristics of ICP-MS, which is widely used in routine analytical applications. The ICP ion source has several unique advantages the samples are introduced at atmospheric pressure the degree of ionisation is relatively uniform for all elements and singly charged ions are the principal ion product. Theoretically, 54 elements can be ionised in an ICP with an efficiency of 90 % or more. Even some elements that do not show ionic emission lines should be ionised with reasonable efficiency (namely, As, 52 % and P, 33%) [381]. This is one of the advantages of ICP-MS over ICP-AES. Other features of ICP-MS that make it more attractive than ICP-AES are much lower detection limits ability to provide isotopic ratio information and to offer isotope dilution capabilities for quantitative analysis and clean and simple spectra. The... 

Isotope dilution (ID) is a technique for the quantitative determination of element concentrations in a sample, on the basis of isotope ratios [382]. An important prerequisite for isotope dilution is the availability of two stable isotopes, although in some cases the use of long-lived radionuclides allows the application range to be further extended [420]. 

Applications The application of the isotope dilution technique is especially useful in carrying out precise and accurate micro and trace analyses. The most accurate results in mass spectrometry are obtained if the isotope dilution technique is applied (RSDs better than 1 % in trace analysis). Therefore, application of IDMS is especially recommended for calibration of other analytical data, and for certification of standard reference materials. The technique also finds application in the field of isotope geology, and is used in the nuclear industry for quantitative isotope analysis. 

Isotope dilution techniques are attractive because they do not require quantitative recovery of the analyte. One must, however, be able to monitor specific isotopes which is possible by using mass spectrometry. 

In the analysis of seawater, isotope dilution mass spectrometry offers a more accurate and precise determination than is potentially available with other conventional techniques such as flameless AAS or ASV. Instead of using external standards measured in separate experiments, an internal standard, which is an isotopically enriched form of the same element, is added to the sample. Hence, only a ratio of the spike to the common element need be measured. The quantitative recovery necessary for the flameless atomic absorption and ASV techniques is not critical to the isotope dilution approach. This factor can become quite variable in the extraction of trace metals from the salt-laden matrix of seawater. Yield may be isotopically determined by the same experiment or by the addition of a second isotopic spike after the extraction has been completed. 

One of the advantages of the isotope dilution technique is that the quantitative recovery of the analytes is not required. Since it is only their isotope ratios that are being measured, it is necessary only to recover sufficient analyte to make an adequate measurement. Therefore, when this technique is used in conjunction with graphite furnace atomic absorption spectrometry, it is possible to determine the efficiency of the preconcentration step. This is particularly important in the analysis of seawater, where the recovery is very difficult to determine by other techniques, since the concentration of the unrecovered analyte is so low. In using this technique, one must assume that isotopic equilibrium has been achieved with the analyte, regardless of the species in which it may exist. 

Quantification is usually achieved by a standard addition method, use of labeled internal standards, and/or external calibration curves. In order to allow for matrix interferences the most reliable method for a correct quantitation of the analytes is the isotope dilution method, which takes into account intrinsic matrix responses, using a deuterated internal standard or carbon-13-labeled internal standard with the same chemistry as the pesticide being analyzed (i.e., d-5 atrazine for atrazine analysis). Quality analytical parameters are usually achieved by participation in interlaboratory exercises and/or the analysis of certified reference materials [21]. 

Isotope Dilution Mass Spectrometry (IDMS) A quantitative mass spectrometry technique in which an isotopically enriched compound is used as an internal standard. See Chapter 14 for a more detailed explanation. 

TI is a very precise and accurate method in stable isotope ratio measurements and quantification of inorganic elements, for example, by isotope dilution mass spectrometry [8]. Because TI is a continuous ion source, it could be coupled to any analyzer that is suitable for such sources. However, because the strength of TI lies in the quantitative precision and accuracy, sector analyzers are preferred to ensure maximum quality. 

Stable-isotope dilution analysis is an analytical technique in which a known quantity of a stable-labelled isotope is added to a sample prior to extraction, in order to quantitate a particular compound. The ratio of the naturally abundant and the stable-labelled isotope is a measure of the naturally abundant compound and can be determined only by gas chromatography-mass spectrometry since the naturally abundant and the stable-labelled isotope cannot be completely separated gas chromatographically. 

Isotope dilution analysis is the definitive method for the quantitative analysis of many compounds but is usually only carried out in specialist laboratories. 

Separation and Quantitation.—The specific radioactivity of [y-32P]ATP of very high activity (up to 240 Ci per millimole) may be measured by using it to phosphorylate [dT(pT)10] quantitatively, using polynucleotide kinase, isolating the labelled undecanucleotide, and measuring its activity.170 Isotope dilution is used to confirm the values. An alternative method of measuring specific radioactivities of ribo-nucleoside triphosphates utilizes a 3H-labelled nucleoside triphosphate (e.g. UTP) of unknown specific activity, a 14C-labelled nucleoside (e.g. ATP), a suitable primer in... 

Calder, A.G. Garden, K.E. Anderson, S.E. Lobley, G.E. Quantitation of Blood and Plasma Amino Acids Using Isotope Dilution GC-EI-MS with U- C Amino Acids As Internal Standards. Rapid Com-mun. Mass Spectrom. 1999, 13, 2080-2083. 

It is critical when performing quantitative GC/MS procedures that appropriate internal standards are employed to account for variations in extraction efficiency, derivatization, injection volume, and matrix effects. For isotope dilution (ID) GC/MS analyses, it is crucial to select an appropriate internal standard. Ideally, the internal standard should have the same physical and chemical properties as the analyte of interest, but will be separated by mass. The best internal standards are nonradioactive stable isotopic analogs of the compounds of interest, differing by at least 3, and preferably by 4 or 5, atomic mass units. The only property that distinguishes the analyte from the internal standard in ID is a very small difference in mass, which is readily discerned by the mass spectrometer. Isotopic dilution procedures are among the most accurate and precise quantitative methods available to analytical chemists. It cannot be emphasized too strongly that internal standards of the same basic structure compensate for matrix effects in MS. Therefore, in the ID method, there is an absolute reference (i.e., the response factors of the analyte and the internal standard are considered to be identical Pickup and McPherson, 1976). 

Jansen, E. E. W., Gibson, K. M., Shigematsu, Y., Jakobs, C., and Verhoeven, N. M. (2006). A novel, quantitative assay for homocamosine in cerebrospinal fluid using stable-isotope dilution liquid chromatography-tandem mass spectrometry. J. Chromatogr. B Arnlyt. Technol. Biomed. Life Sci. 830,196-200. 

The classical methods for detection and quantitation of racemization require analysis of the chiral purity of the product of a peptide-bond-forming reaction. For example, the Anderson test is used to explore a variety of reaction conditions for the coupling of Z-Gly-Phe-OH to H-Gly-OEt (Scheme 6). 9 The two possible enantiomeric tripeptides are separable by fractional crystallization, so that gravimetric analysis furnishes the racemization data. This procedure has a detection limit of 1-2% of the epimerized tripeptide. A modification by Kemp,1"1 utilizing 14C-labeled carboxy components, extends the detection limit by two to three orders of magnitude by an isotopic dilution procedure. The Young test 11 addresses the coupling of Bz-Leu-OH to H-Gly-OEt, and the extent of epimerization is determined by measurement of the specific rotation of the dipeptide product. 

The isotope dilution method consists of mixing a natural sample with an artificial spike and measuring the isotopic ratios of the mixture using mass spectrometry, providing very precise quantitative determination of the concentrations of elements in trace quantities. A spike is a solution that contains a known concentration of the element, artificially enriched in one of its minor isotopes. For example, natural Rb samples have 27.84% and 72.16% Rb (Fig. 11.lA). Rb spikes are made by artificially enrich the minor isotope Rb. And a solution with 90% Rb and 10% Rb (Fig. 11.IB) is a Rb spike. Of course, a solution with 99.99% Rb and 0.01% Rb is a better Rb spike. When the known quantities (mass) of the sample and spike are mixed, the resulting isotopic compositions can be used to calculate the concentration of the element in the sample. 

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