Isotopic Analogue Research Articles

Combination of quadrupole isotope dilution mass spectrometry with simultaneous derivatization and spray assisted droplet formation-liquid phase microextraction for the determination of methamphetamine in human urine and serum samples by gas chromatography mass spectrometry

In this study, an analytical method with high accuracy and precision was developed for the determination of methamphetamine in human urine and serum samples by gas chromatography–mass spectrometry (GC–MS). A simultaneous derivatization and spray assisted droplet formation-liquid phase microextraction (SADF-LPME) method was proposed to derivatize and preconcentrate target analyte. Quadruple isotope dilution (ID4) was used to provide high accuracy and precision for methamphetamine determination in the samples. After the optimization studies for the derivatization and microextraction parameters, limit of detection (LOD) and limit of quantitation (LOQ) for the developed SADF-LPME method were found to be 48.0 and 159.9 μg/kg, respectively. Recovery studies were implemented to verify the applicability and accuracy of the developed method for human urine and serum samples. The SADF-LPME method gave low percent recovery results (30.5–61.0%) for the spiked urine and serum samples showing that it failed to minimize or eliminate matrix effects for the analyte. Hence, methamphetamine acetamide-d3 was synthesized and purified in our research laboratory to be used as methamphetamine isotopic analogue in the ID4 method. When the SADF-LPME method was combined with ID4, the percent recovery values for urine and serum samples were calculated as 99.7–100.0% and 99.4–100.2%, respectively. These results demonstrated the applicability and accuracy of the proposed method for urine and serum samples.

Quadruple isotope dilution gas chromatography-mass spectrometry after simultaneous derivatization and spraying based fine droplet formation liquid phase microextraction method for the accurate and sensitive quantification of chloroquine phosphate in human serum, urine and saliva samples at trace levels

This study presents an accurate and precise analytical strategy for the determination of chloroquine phosphate at trace levels in human body fluids (urine, serum, and saliva). Simultaneous derivatization-spraying based fine droplet formation-liquid phase microextraction (SD-SFDF-LPME) method was used to derivatize and preconcentrate the analyte prior to gas chromatography-mass spectrometry (GC–MS) measurements. Acetic anhydride was employed as derivatizing agent in this study. After optimizing the SD-SFDF-LPME method, the limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.16 and 0.53 mg/kg, respectively. Quadruple isotope dilution (ID4) was coupled to the SD-SFDF-LPME method in order to alleviate matrix effects and promote accuracy/precision of the method. Chloroquine acetamide-d3 was firstly synthesized in our research laboratory and used as the isotopic analogue of the analyte in the ID4 experiments. Superior percent recovery results (99.4% – 101.0%) with low standard deviation values were obtained for the spiked samples. This validated the developed SD-SFDF-LPME-ID4-GC–MS method as highly accurate and precise for the determination of chloroquine phosphate at trace levels. In addition, the isotopic analogue of the analyte was obtained via the acetamide derivative of the analyte, which is an alternative to obtain isotopic analogues of organic compounds that are not accessible or commercially available.

Muonium Chemistry at Diiron Subsite Analogues of [FeFe]-Hydrogenase.

The chemistry of metal hydrides is implicated in a range of catalytic processes at metal centers. Gaining insight into the formation of such sites by protonation and/or electronation is therefore of significant value in fully exploiting the potential of such systems. Here, we show that the muonium radical (Mu.), used as a low isotopic mass analogue of hydrogen, can be exploited to probe the early stages of hydride formation at metal centers. Mu. undergoes the same chemical reactions as H. and can be directly observed due to its short lifetime (in the microseconds) and unique breakdown signature. By implanting Mu. into three models of the [FeFe]‐hydrogenase active site we have been able to detect key muoniated intermediates of direct relevance to the hydride chemistry of these systems.

Muonium Chemistry at Diiron Subsite Analogues of [FeFe]‐Hydrogenase

AbstractThe chemistry of metal hydrides is implicated in a range of catalytic processes at metal centers. Gaining insight into the formation of such sites by protonation and/or electronation is therefore of significant value in fully exploiting the potential of such systems. Here, we show that the muonium radical (Mu.), used as a low isotopic mass analogue of hydrogen, can be exploited to probe the early stages of hydride formation at metal centers. Mu. undergoes the same chemical reactions as H. and can be directly observed due to its short lifetime (in the microseconds) and unique breakdown signature. By implanting Mu. into three models of the [FeFe]‐hydrogenase active site we have been able to detect key muoniated intermediates of direct relevance to the hydride chemistry of these systems.

Geological, petrologic, isotopic, and geochemical constraints of geodynamic models simulating formation of the archean tonalite-trondhjemite-granodiorite associations in ancient cratons

The geological setting, geochemistry, and Nd isotopic systematics of tonalite-trondhjemite-granodiortite (TTG) series in ancient cratons are considered. It is shown that the TTG series were formed from ∼4.2 to 2.6 Ga ago in the oldest continental cores; many TTG series do not reveal chronological links to greenstone belts. This follows from the evolution of the Slave Craton in the Canadian Shield, the Vodlozero Craton in the Baltic Shield, and the Pilbara and Yilgarn cratons in the Australian Shield, where greenstone associations postdated TTG series. As has been established at the Baltic Shield, the primary melts of the Mesoarchean TTG associations were formed at a shallower depth (P 150 Ma) that separates the formation of the TTG melts from the age of the source involved in melting. Taking into account the calculated rate of cooling of the lithospheric plates, these data indicate that most Archean TTG series likely were not formed in the convergent subduction-related and accretionary geodynamic settings. The isotopic and geochemical data constrain compositions of the sources that produced Archean TTG series. Petrologic modeling of the formation conditions and Nd isotopic composition of the metabasalts in greenstone belts show that these rocks could not have been the source of TTG series. The most plausible isotopic and geochemical analogue of this source are the Archean amphibolites (ENd mafic rocks), which differ from the metabasalts of greenstone belts by their lower Sm/Nd ratio and enrichment in some lithophile elements. The available data suggest that the primary TTG melts were generated as products of melting of amphibolites and granulites forming the lower crust.

Measurement of progesterone in human serum by isotope dilution liquid chromatography–tandem mass spectrometry and comparison with the commercial chemiluminescence immunoassay

Progesterone is one of the steroid hormones. The hormone is especially important in preparing the uterus for the implantation of the blastocyst and in maintaining pregnancy. Its concentration in serum is measured to determine ovarian function and to predict early pregnancy. The progesterone concentration is also important for in-vitro fertilization and embryo-transfer outcomes. We have established isotope dilution liquid chromatography-tandem mass spectrometry as a primary method for the measurement of progesterone in human serum. Progesterone and its isotopic analogue, progesterone-(13)C(2), in serum were monitored at mass transitions of m/z 315.2/109.2 and 317.2/111.2 respectively in multiple-reaction monitoring (MRM) mode with electrospray positive ionization. For validation of the method, progesterone in a National Institute of Standards and Technology standard reference material (NIST SRM) was measured, and the measured results were in good agreement with the reference values within the uncertainty. On the basis of the established method, progesterone certified reference material (CRM) was developed in this work. The certified value was (1.41 ± 0.036) μg kg(-1). The repeatability of 1.1% and reproducibility of 0.14% showed that ID LC-MS-MS is a reliable and reproducible method. The expanded uncertainty for the measurement of progesterone in the CRM was approximately 2.6% within 95% confidence limits. The detection limit of progesterone was approximately 0.6 μg kg(-1). The progesterone CRMs were distributed to representative clinical laboratories in the Republic of Korea for comparison with the chemiluminescence immunoassay (CLIA), which is the most sensitive immunoassay method. The results from the comparison showed quite a large bias among the participating laboratories. This implies that the CRM is a very important material for establishment of traceability to its practical use.

Determination of phenylalanine in human serum by isotope dilution liquid chromatography/tandem mass spectrometry

Isotope dilution liquid chromatography/tandem mass spectrometry (ID-LC/MS/MS) has been developed as a candidate reference method to determine the level of phenylalanine in human serum. The advantages of this method include a simple sample preparation without derivatization, selective detection of analytes, and the use of an isotopic analogue as an internal standard. Phenylalanine and its isotopic analogue, phenylalanine-ring-(13)C(6), were monitored at the transitions m/z 166.2/120.2 and 172.2/126.2 in the multiple-reaction monitoring (MRM) mode, respectively. The expanded uncertainty of the measurement result of phenylalanine in the serum was approximately 1.2% within a 95% confidence level. A standard reference material, with a certified value of phenylalanine, was analyzed in order to verify this method. The result obtained by the ID-LC/MS/MS method differed somewhat from the certified value, but agreed well with the gravimetric value. The measurement result of phenylalanine in serum by ID-LC/MS/MS was compared with the results from the commercial HPLC method, which was carried out in clinics. The results from the commercial HPLC method showed inconsistent results with each other. The busted results from the commercial HPLC method suggest that it should be possible to trace the results of the commercial fields to well-characterized reference materials or methods.

A novel 10B-enriched carboranyl-containing phthalocyanine as a radio- and photo-sensitising agent for boron neutron capture therapy and photodynamic therapy of tumours: in vitro and in vivo studies

The synthesis of a Zn(ii)-phthalocyanine derivative bearing four 10B-enriched o-carboranyl units (10B-ZnB4Pc) and its natural isotopic abundance analogue (ZnB4Pc) in the peripheral positions of the tetraazaisoindole macrocycle is presented. The photophysical properties of ZnB4Pc, as tested against model biological systems, were found to be similar with those typical of other photodynamically active porphyrin-type photosensitisers, including a singlet oxygen quantum yield of 0.67. The carboranyl-carrying phthalocyanine was efficiently accumulated by B16F1 melanotic melanoma cells in vitro, appeared to be partitioned in at least some subcellular organelles and, upon red light irradiation, induced extensive cell mortality. Moreover, ZnB4Pc, once i.v.-injected to C57BL/6 mice bearing a subcutaneously transplanted pigmented melanoma, photosensitised an important tumour response, provided that the irradiation at 600-700 nm was performed 3 h after the phthalocyanine administration, when appreciable concentrations of ZnB4Pc were still present in the serum. Analogously, irradiation of the 10B-ZnB4Pc-loaded pigmented melanoma with thermal neutrons 24 h after injection led to a 4 day delay in tumour growth as compared with control untreated mice. These results open the possibility to use one chemical compound as both a photosensitising and a radiosensitising agent for the treatment of tumours by the combined application of photodynamic therapy and boron neutron capture therapy.

Giant Isotope Effect in the Incoherent Tunneling Specific Heat of the Molecular NanomagnetFe8

Time-dependent specific heat experiments on the molecular nanomagnet Fe8 and the isotopic enriched analogue 57Fe8 are presented. The inclusion of the 57Fe nuclear spins leads to a huge enhancement of the specific heat below 1 K, ascribed to a strong increase in the spin-lattice relaxation rate gamma arising from incoherent, nuclear-spin-mediated magnetic quantum tunneling (MQT) in the ground doublet. Since gamma is found comparable to the expected tunneling rate, the MQT process has to be inelastic. A model for the coupling of the tunneling spins to the lattice is presented. Under transverse field, a crossover from nuclear-spin-mediated to phonon-induced tunneling is observed.

Hyperfine and Host−Guest Interactions of the Mu-Cyclohexadienyl Radical in NaY Zeolite

The adsorption and dynamical behavior of the muonated cyclohexadienyl radical (C6H6Mu) in NaY zeolite, formed by muonium (Mu) addition on adsorbed benzene, was investigated by the muon spin resonance (μSR) technique, primarily at loadings of 2−3 C6H6 molecules per supercage of NaY. The dynamics of this radical are expected to be the same as its isotopic analogue, C6H7, for which there are no similar data available. Both TF-μSR and ALC-μSR spectra were recorded, with the most detailed information provided by the positions and line widths of the avoided level crossing resonances. In concert with 2H NMR, neutron diffraction and molecular dynamics studies of the parent benzene molecule, as well as current theoretical calculations, the dominant adsorption site for the C6H6Mu radical is believed to be the SII Na cation, within a supercage, which gives rise to three observed ALC lines, corresponding to two different orientations for the muon (proton) of the CHMu methylene group: pointing toward (endo) and away ...

Performance characteristics and applications of a proton transfer reaction-mass spectrometer for measuring volatile organic compounds in ambient air.

Data illustrating the performance characteristics of a proton transfer reaction-mass spectrometer (PTR-MS) under both laboratory and field conditions are presented. Under laboratory conditions, we demonstrate that PTR-MS measures (within 10%) a 2.6 ppbv concentration of gaseous dimethyl sulfide. Using a stepwise dilution of a gaseous isoprene standard, we demonstrate the linearity of the response of PTR-MS across 3 orders of magnitude of mixing ratios, from 100 ppbv to less than 100 pptv. By combining this data set with that of its monosubstituted 13C isotopic analogue, we demonstrate the ability of the instrumentto reliably measure concentrations as low as approximately 50 pptv and to detect concentrations at significantly lower levels. We conclude our laboratory characterization by investigating the components of the instrument noise signal (drift, mean, and range) and develop an expression (noise statistic) that reliably predicts the instrumental noise associated with any signal across a wide range of masses. In the field, we deployed a PTR-MS at a clean-air coastal site and an urban kerbside monitoring station to demonstrate the measurement of atmospheric dimethyl sulfide and benzene concentrations, respectively. At both sites, we were able to monitor diurnal variations in concentrations at unprecedented temporal resolutions (<5 min between successive measurements). We then demonstrate how the noise statistic can be applied to enable real fluctuations in atmospheric VOC concentrations to be reliably distinguished from instrument noise. We conclude by demonstrating how PTR-MS can be used to measure real-time VOC emission rate changes from vegetation in response to external forcing by examining the effect varying photon-flux density has upon emissions of isoprene from a Sitka spruce tree.

Intracellular Ca 2+ changes induced by D 2O/H 2O solvent isotope exchange in devacuolated internodal cells of Chara gymnophylla

Deuterium oxide (D 2O) an isotopic analogue of water is known to cause membrane excitation through activation of Ca 2+ channels in giant internodal cells of the freshwater alga Chara gymnophylla. Internally perfused devacuolated and ligated internodal cells loaded with fura-2 and overloaded with Ca 2+ (pathologic or hyperexcited condition) or with normal (resting) cytosolic Ca 2+ concentration ([Ca 2+] i) were used to study the effect of D 2O/H 2O solvent isotope exchange on Ca 2+ homeostasis. In spite of internal perfusion, devacuolated internodal cells showed significant cytosolic Ca 2+-buffering capacity that was sensitive to external calcium and sodium azide. D 2O application induced a rise in [Ca 2+] i preceded by an early transient decrease. With respect to the amplitude and rate of the [Ca 2+] i rise, as well as to the magnitude of early transient decay, the responses of different preparations showed a bimodal distribution (L- or S-type responses). The early transient [Ca 2+] i decrease was not affected by the Ca 2+ channel inhibitor, nicardipine, but the cation channel blocker, La 3+ abolished it. The [Ca 2+] i rising phase in L-type cells was partly channel mediated (dependent on external Ca 2+, La 3+ and nicardipine) and could be compared to the [Ca 2+] i rise induced by KCl. Applying a hyperosmotic medium mimicked the observed D 2O-induced [Ca 2+] i changes, however the time-course was significantly different. The fraction of the [Ca 2+] i rise manifested in S- type cells as well as after Ca 2+ channel block or in Ca 2+-free media, revealed a possible novel component of D 2O-induced excitation ascribed to residual Ca 2+ stores.

Determination of specific radioactivity for 76br-labeled compounds measuring the ratio between 76br and 79br using packed capillary liquid chromatography mass spectrometry

Packed capillary liquid chromatography with electrospray mass spectrometry was used for direct determination of the specific radioactivity by calculation of isotope ratios between the 76Br- and 79Br-labeled analogues of N-((3-aminomethyl)benzyl)-4-bromobenzamide. Using 20 μL injections on packed capillary columns, sufficient mass sensitivity was attained for the determination on an injected amount of radioactivity corresponding to approximately 2 MBq (0.3 pmol of the 76Br isotopic analogue).

Chemical derivatization and the selection of deuterated internal standard for quantitative determination--methamphetamine example.

Use of an isotopic analogue of the analyte as the internal standard in a quantitative gas chromatography-mass spectrometry targeted-compound-analysis protocol is often hindered by the availability of an adequate number (typically three for the drug/metabolite and two for the isotopic analogue) of sufficiently high mass ions that can be attributed to each member of the pair and are sufficiently free of interference by the contribution from the other component of the pair, a phenomenon termed "cross-contribution". Methamphetamine (MA) is selected as the exemplar compound to examine the effectiveness in using different chemical derivatization routes to produce derivatized analyte-isotopic analogue pairs that can generate more favorable mass spectrometric data to meet this analytical requirement. Trimethylsilyl-, trichloroacetyl-, and pentafluoropropionyl-derivatization and MA-d5, MA-d8, and MA-d9 are studied. Data resulting from this study indicate that the number of ion pairs suitable for quantitation and the degree of cross-contribution of these ions vary significantly. These data empirically demonstrate that derivatization methods play a significant role in deciding which deuterated analogue of the analyte provides the most suitable ion pairs that cause the least cross-contribution. The most suitable internal standard varies with the derivatization route adapted for an analytical protocol.

Variable Magnetism of Layer-Structured Compounds Cu2(OD)3X with Exchangeable Anion X: Magnetic Local Structure and Magnetic Interactions Determined by Solid-State High-Resolution Deuterium NMR

The microscopic magnetic local structure of Botallackite-type layer-structured compounds Cu2(OD)3X (X = C6H5COO−) exhibiting a nonequilateral planar triangular copper lattice was determined by the solid-state high-resolution deuterium NMR above 190 K. The magnetic interaction in a copper layer was probed by the isotropic NMR shifts of OD− groups and could be approximated by a sum of 1D-Heisenberg chains in the high temperature region. Two copper chains with different exchange interactions J = −54 ± 4 and −92 ± 3 K were found for Cu2(OD)3C6H5COO from the temperature dependence of the three distinct NMR signals. The derived values of J almost reproduced the high temperature behavior of the magnetic susceptibility χA T vs. T. The magnetic susceptibility of isotopic analogue Cu2(OH)3C6D5COO showed ferromagnetic interaction below 50 K. Electron spin distribution in a phenyl ring of a benzoate anion is governed by spin polarization mechanism and it's magnitude is very small. Magnetic interaction between the layers through the benzoate anion is very weak.

Carbon acceptors passivated with hydrogen and the search for carbon donors in highly doped GaAs:C

Highly carbon doped p-type GaAs grown by MOVPE shows infrared absorption lines from localized vibrational modes of CAs acceptors and passivated H-CAs pairs. A comparison with samples grown by MOMBE shows that an unassigned line at 563 cm-1 is due to a mode of H-CAs pairs, although an isotopic analogue from D-CAs pairs in plasma-treated samples has not been detected. The important new result is the demonstration that the line cannot be due to CGa donors, and so there is still no evidence for the amphoteric behaviour of carbon impurities in GaAs.

Stable isotope tracer analysis by GC-MS, including quantification of isotopomer effects.

In metabolic tracer studies it is frequently useful to infuse tracers that are differently labeled variants of the same molecule. These tracers are known as isotopomers. Analysis of the enrichment of each isotopic analogue can be accomplished by gas chromatography-mass spectrometry (GC-MS). However, the raw GC-MS data must be corrected to give the information required. This paper addresses how to transform the raw GC-MS data, consisting of relative abundance ratios at specific ion masses, into relative molar ratios of tracer and tracee molecules. Several correction factors are necessary. First, the background must be measured and corrected for, since it is always present in the sample. Second, the abundances in the spectrum of the labeled molecule are different from those in the unlabeled molecule, and this proportionality "skew" is corrected. A third correction factor accounts for the overlapping spectra of two or more isotopomers that cannot be measured independently. The final correction removes the "double vision" effect that may appear in some spectra due to the presence of (M - H)+ species.

Isotopic Analogue as the Internal Standard for Quantitative Determination of Benzoylecgonine: Concerns with Isotopic Purity and Concentration Level

Abstract Empirical data is used to demonstrate the observation and quantification of benzoylecgonine in negative test samples when high concentrations of deuterated benzoylecgonine are used as the internal standard in the assay process. On the quantitative determination of true positive samples, inaccuracy introduced by the isotopic impurity of the internal standard is calculated as a function of the impurity and the concentration levels of the internal standard used.

Effects of isotopic substitution on the vibrational spectra of 2,4,6-trinitrotoluene

The gas phase FT-i.r. spectra of 2,4,6-trinitrotoluene and various 2H and 15N substituted analogues are recorded at temperatures above the melting point of each compound. AM1 calculations are used to predict the fundamental vibrational frequencies of each isotopic analogue. Solid state (KBr) spectra and solvent effects aided in the vibrational assignments.