Determination of Amino Acid Isotope Enrichment Using Liquid Chromatography–Mass Spectrometry

Eijk, Hans M.H. van; Rooyakkers, D.R.; Soeters, Peter B.; Deutz, N.E.P.

doi:10.1006/abio.1999.4112

Cited by 89 publications

(83 citation statements)

References 18 publications

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“…Our method uses standard compounds to make a calibration curve and therefore is independent of integration algorithm, concentration effect in ionization method or detection system. Accordingly, it is applicable to most instruments such as Iron-Trap, [46] Tof, [17,20] sector or Fourier transform-ion cyclotron resonance (FT-ICR MS) [18] systems, as well as separation-detection systems like liquid chromatography (LC)-MS [19] or capillary electrophoresis (CE)-MS in reason that the accuracy of calibration curve relies only on the accuracy of predetermined MIDs of the biosynthetic compounds, (3) since the method can generate labeled compounds with specific C j (any/each), it could be used for assessment of the data accuracy of any instrument. In the work of Van Eijk et al, measurement of isotopomer fraction by LC-MS was reported, but not its accuracy.…”

Section: Calibration Curves Using Biosynthetic Amino Acidsmentioning

confidence: 99%

“…In the work of Van Eijk et al, measurement of isotopomer fraction by LC-MS was reported, but not its accuracy. [19] Jeffrey et al has firstly employed the tandem mass spectrometry to acquire more constraints of mass isotopomers, but the accuracy of the result was not surveyed neither. [47] Further, plenty of MS data from tracer studies are reported without any indication of error.…”

Section: Calibration Curves Using Biosynthetic Amino Acidsmentioning

confidence: 99%

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Increasing the accuracy of mass isotopomer analysis through calibration curves constructed using biologically synthesized compounds

Shen

Xiong

et al. 2009

J. Mass Spectrom.

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Mass isotopomer analysis is an important technique to measure the production and flow of metabolites in living cells, tissues, and organisms. This technique depends on accurate quantifications of different mass isotopomers using mass spectrometry. Constructing calibration curves using standard samples is the most universal approach to convert raw mass spectrometry measurements into quantitative distributions of mass isotopomers. Calibration curve approach has been, however, of very limited use in comprehensive analyses of biological systems, mainly suffering from the lack of extensive range of standard samples with accurately known isotopic enrichment. Here, we present a biological method capable of synthesizing specifically labeled amino acids. These amino acids have well-determined and estimable mass isotopomer distributions and thus can serve as standard samples. In this method, the bacterium strain Methylobacterium salsuginis sp. nov. was cultivated with partially 13 C-labeled methanol as the only carbon source to produce 13 C-enriched compounds. We show that the mass isotopomer distributions of the various biosynthesized amino acids are well determined and can be reasonably estimated based on proposed binomial approximation if the labeling state of the biomass reached an isotopic steady state. The interference of intramolecular inhomogeneity of 13 C isotope abundances caused by biological isotope fractionation was eliminated by estimating average 13 C isotope abundance. Further, the predictions are tested experimentally by mass spectrometry (MS) spectra of the labeled glycine, alanine, and aspartic acid. Most of the error in mass spectrometry measurements was less than 0.74 mol% in the test case, significantly reduced as compared with uncalibrated results, and this error is expected to be less than 0.4 mol% in real experiment as revealed by theoretical analysis.

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Section: Calibration Curves Using Biosynthetic Amino Acidsmentioning

confidence: 99%

Section: Calibration Curves Using Biosynthetic Amino Acidsmentioning

confidence: 99%

Increasing the accuracy of mass isotopomer analysis through calibration curves constructed using biologically synthesized compounds

Shen

Xiong

et al. 2009

J. Mass Spectrom.

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“…Approximately 80 mg of tissue were added to 400 l of 5% sulfosalicylic acid (SSA), with 300 mg of glass beads, and beaten for 30 s (8). Plasma and tissue amino acid concentrations and enrichments were measured with a fully automated liquid chromatography-mass spectrometry system, using precolumn derivatization with o-phthalaldehyde (32 H2]tyrosine, using the steady-state isotope dilution equation…”

Section: Experimental Protocolmentioning

confidence: 99%

Renal arginine and protein synthesis are increased during early endotoxemia in mice

Hallemeesch

Soeters

Deutz

2002

American Journal of Physiology-Renal Physiology

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Hallemeesch, Marcella M., Peter B. Soeters, and Nicolaas E. P. Deutz. Renal arginine and protein synthesis are increased during early endotoxemia in mice. Am J Physiol Renal Physiol 282: F316-F323, 2002. First published August 21, 2001 10.1152/ajprenal.00039.2001The kidney has an important function in arginine metabolism, because the kidney is the main endogenous source for de novo arginine production from circulating citrulline. In conditions such as sepsis, nitric oxide (NO) production is increased and is dependent on extracellular arginine availability. To elucidate the adaptive role of renal de novo arginine synthesis in a condition of increased NO production, we studied renal arginine metabolism in a mouse model of endotoxemia. Because arginine flux is largely dependent on protein flux, we also measured protein metabolism in mice. Female mice were injected intraperitoneally with lipopolysaccharide; control mice received 0.9% NaCl. Six hours later, renal blood flow was measured with the use of para-aminohippuric acid. Arginine and protein metabolism were studied using organbalance, stable-isotope techniques. Systemic NO production was increased in the endotoxin-treated mice. In addition, renal protein synthesis and de novo arginine production from citrulline were increased. However, no effect on renal NO production was observed. In conclusion, increased renal de novo arginine production may serve to sustain systemic NO production. To our knowledge, it was shown for the first time that renal protein synthesis is enhanced in the early response to endotoxemia. citrulline; kidney; lipopolysaccharide; nitric oxide THE AMINO ACID ARGININE HAS an important role in cellular regeneration, immune function, and protein breakdown and synthesis (7,39). Arginine also is the precursor of urea, agmatine, and nitric oxide (NO). In past years, many studies have been undertaken that aimed at increasing or decreasing NO production, either by supplementation of arginine or NO donors (6,25,26) or by the administration of nitric oxide synthase (NOS) inhibitors or arginase (5, 15,33). In general, the effects of these treatments were conflicting and confusing. More detailed information on local arginine metabolism and NO production may be helpful for an understanding of the effects of arginine supplementation or NOS inhibition.The kidney has an important function in arginine metabolism, because the kidney is the main endogenous source for de novo arginine production (37). Arginine production involves collaboration between the gut and the kidney. Citrulline is produced from glutamine and arginine in the gut and then released into the circulation. The liver does not consume citrulline, which leaves it available for the kidney. In the kidney, citrulline is used for the production of arginine, which, in turn, is released into the circulation (37). The importance of this pathway is illustrated by the fact that arginine becomes an essential amino acid when intestinal citrulline synthesis is inhibited (34).During sepsis and experimental ...

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“…Overall, the results presented in Table 4 compare favorably with similar work for derivatized amino acids [34][35][36]. These studies used ESI with either ion trap or QMS, as well as fast atom bombardment ionization-QMS.…”

Section: Comparison Of the Precision And Accuracy Of The Sim And Scanmentioning

confidence: 59%

“…This result suggests the method can detect small levels of enrichment, even for very low natural abundance levels. The ability to detect such low enrichment may be extremely valuable for measuring the incorporation of isotopically enriched amino acids into proteins [31][32][33].Overall, the results presented in Table 4 compare favorably with similar work for derivatized amino acids [34][35][36]. These studies used ESI with either ion trap or QMS, as well as fast atom bombardment ionization-QMS.…”

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confidence: 60%

Inaccuracies in selected ion monitoring determination of isotope ratios obviated by profile acquisition: nucleotide 18O/16O measurements

Cassano

Wang

Anderson

et al. 2007

Analytical Biochemistry

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Precise and accurate measurements of isotopologue distributions (IDs) in biological molecules are needed for the determination of isotope effects, quantitation by isotope dilution and quantifying isotope tracers employed in both metabolic and biophysical studies. While single ion monitoring (SIM) yields significantly greater sensitivity and signal/noise than profile mode acquisitions, we show that small changes in the SIM window width and/or center can alter experimentally determined isotope ratios by up to 5%, resulting in significant inaccuracies. This inaccuracy is attributed to mass granularity, the differential distribution of digital data points across the m/z ranges sampled by SIM. Acquiring data in the profile mode and fitting the data to an equation describing a series of equally spaced and identically shaped peaks eliminates the inaccuracies associated with mass granularity with minimal loss of precision. Additionally a method of using the complete ID profile data that inherently corrects for "spillover" and for the natural abundance ID has been used to determine 18 Keywords isotope ratio; isotopologue distribution; mass isotopomer distribution; tandem mass spectrometry; heavy atom isotope effect; oxygen-18; nucleotide; mass granularity; whole molecule mass spectrometry 1 This work supported by NIH grants GM56740 (M.E.H.), R33 DK07029 (VEA & SP) and a HHMI predoctoral fellowship (A.G.C.). *Corresponding Authors: Vernon E. Anderson and Michael E. Harris, Department of Biochemistry, Case Western Reserve University School of Medicine, Cleveland, OH 44106-4935, Phone: 216-368-2511 (VEA); 216-368-4779 (MEH), Fax: 216-368-3419, Email: vernon.anderson@case.edu; michael.harris@cwru.edu. 3 Current address: Department of Chemistry, Drew University, Publisher's Disclaimer: This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. NIH Public Access Author ManuscriptAnal Biochem. Author manuscript; available in PMC 2008 August 1. Published in final edited form as:Anal Biochem. 2007 August 1; 367(1): 28-39. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author ManuscriptHeavy atom isotope effects generated by substituting 13 C, 15 N or 18 O are important tools for probing the mechanisms and transition states of chemical reactions [1], including enzymecatalyzed reactions [2][3][4][5][6][7]. Because the observed effects for isotopic substitution of heavy atoms are extremely small, typically a few percent, they cannot be determined by direct comparison of experimentally determined rate constants. Instead, heavy atom isotope effects are generally measured by internal competition methods that...

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Determination of Amino Acid Isotope Enrichment Using Liquid Chromatography–Mass Spectrometry

Cited by 89 publications

References 18 publications

Increasing the accuracy of mass isotopomer analysis through calibration curves constructed using biologically synthesized compounds

Increasing the accuracy of mass isotopomer analysis through calibration curves constructed using biologically synthesized compounds

Renal arginine and protein synthesis are increased during early endotoxemia in mice

Inaccuracies in selected ion monitoring determination of isotope ratios obviated by profile acquisition: nucleotide 18O/16O measurements

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