Nuclear magnetic resonance spectroscopy as a quantitative tool to determine the concentrations of biologically produced metabolites: implications in metabolites in safety testing.


:Nuclear magnetic resonance (NMR) spectroscopy has traditionally been considered as an indispensable tool in elucidating structures of metabolites. With the advent of Fourier transform (FT) spectrometers, along with improvements in software and hardware (such as high-field magnets, cryoprobes, versatile pulse sequences, and solvent suppression techniques), NMR is increasingly being considered as a critical quantitative tool, despite its lower sensitivity as compared to mass spectrometry. A specific quantitative application of NMR is in determining the concentrations of biologically isolated metabolites, which could potentially be used as reference standards for further quantitative work by liquid chromatography/mass spectrometry. With the recent demands from regulatory agencies on quantitative information on metabolites, it is proposed that NMR will play a significant role in strategies aimed at addressing metabolite coverage in toxicological species. Traditionally, biologically isolated metabolites have not been considered as a way of generating "reference standards" for further quantitative work. However, because of the recent FDA guidance on safety testing of metabolites, one has to consider means of authenticating and quantitating biologically or nonbiologically generated metabolites. 1H NMR is being proposed as the method of choice, as it is able to be used as both a qualitative and a quantitative tool, hence allowing structure determination, purity check, and quantitative measurement of the isolated metabolite. In this publication, the application of NMR as a powerful and robust analytical technique in determining the concentrations of in vitro or in vivo isolated metabolites is discussed. Furthermore, to demonstrate the reliability and accuracy of metabolite concentrations determined by NMR, validation and cross-validation with gravimetric and mass spectrometric methods were conducted.


Chem Res Toxicol


Espina R,Yu L,Wang J,Tong Z,Vashishtha S,Talaat R,Scatina J,Mutlib A




Has Abstract


2009-02-01 00:00:00












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    abstract::The growth-promoting effects of estrogens in hormone-dependent tumor tissues involve receptor-mediated pathways that are well-recognized; however, the role of estrogens in tumor initiation remains controversial. Estrogen metabolites, primarily the catechol estrogens (CE's), have been implicated in tumor initiation via...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Mobley JA,Bhat AS,Brueggemeier RW

    更新日期:1999-03-01 00:00:00

  • Effect of aminoglutethimide on neutrophils in rats: implications for idiosyncratic drug-induced blood dyscrasias.

    abstract::Aminoglutethimide (AMG) is an aromatic amine aromatase inhibitor associated with a high incidence of idiosyncratic blood dyscrasias, especially agranulocytosis. Animal models of idiosyncratic drug reactions (IDRs) represent essential tools to study these reactions; however, there is currently no valid model of idiosyn...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Ng W,Uetrecht J

    更新日期:2013-08-19 00:00:00

  • Metabonomic evaluation of Schaedler altered microflora rats.

    abstract::Previously, we identified two distinct metabonomic phenotypes in Sprague-Dawley rats sourced from two different rooms (colonies) in the Charles River, Raleigh facility [Robosky, L. C., Wells, D. F., Egnash, L. A., Manning, M. L., Reily, M. D., and Robertson, D. G. (2005) Metabonomic identification of two distinct phen...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Rohde CM,Wells DF,Robosky LC,Manning ML,Clifford CB,Reily MD,Robertson DG

    更新日期:2007-10-01 00:00:00

  • Cytotoxicity of calcium rectorite micro/nanoparticles before and after organic modification.

    abstract::Organically modified rectorite (OREC) micro/nanoparticles can be synthesized by organic modification from calcium rectorite (Ca(2+)-REC or REC), a common form of rectorite in nature. Although REC and OREC have potential applications in food packing and drug delivery, their cytotoxicity is not clear. In the present stu...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Liu Y,Deng H,Xiao C,Xie C,Zhou X

    更新日期:2014-08-18 00:00:00

  • Covalent Modification of CDK2 by 4-Hydroxynonenal as a Mechanism of Inhibition of Cell Cycle Progression.

    abstract::Oxidative stress is a contributing factor in a number of chronic diseases, including cancer, atherosclerosis, and neurodegenerative diseases. Lipid peroxidation that occurs during periods of oxidative stress results in the formation of lipid electrophiles, which can modify a multitude of proteins in the cell. 4-Hydrox...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Camarillo JM,Rose KL,Galligan JJ,Xu S,Marnett LJ

    更新日期:2016-03-21 00:00:00

  • Human flavin-containing monooxygenase form 2 S-oxygenation: sulfenic acid formation from thioureas and oxidation of glutathione.

    abstract::Thioureas are oxygenated by flavin-containing monooxygenases (FMOs), forming reactive sulfenic and/or sulfinic acids. Sulfenic acids can reversibly react with GSH and drive oxidative stress through a redox cycle. For this reason, thiourea S-oxygenation is an example of FMO-dependent bioactivation of a xenobiotic. Func...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Henderson MC,Krueger SK,Stevens JF,Williams DE

    更新日期:2004-05-01 00:00:00

  • Preclinical strategy to reduce clinical hepatotoxicity using in vitro bioactivation data for >200 compounds.

    abstract::Drug-induced liver injury is the most common cause of market withdrawal of pharmaceuticals, and thus, there is considerable need for better prediction models for DILI early in drug discovery. We present a study involving 223 marketed drugs (51% associated with clinical hepatotoxicity; 49% non-hepatotoxic) to assess th...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Sakatis MZ,Reese MJ,Harrell AW,Taylor MA,Baines IA,Chen L,Bloomer JC,Yang EY,Ellens HM,Ambroso JL,Lovatt CA,Ayrton AD,Clarke SE

    更新日期:2012-10-15 00:00:00

  • A new role for glutathione: protection of vitamin B12 from depletion by xenobiotics.

    abstract::NADPH in microsomes reduces the hydroxocob(III)alamin form of vitamin B12 to cob(II)alamin and the supernucleophilic cob(I)alamin, which are both highly reactive toward xenobiotic epoxides formed by mammalian metabolism of dienes such as the industrially important chemicals chloroprene and 1,3-butadiene. With styrene,...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Watson WP,Munter T,Golding BT

    更新日期:2004-12-01 00:00:00

  • Reaction of lysine with estrone 3,4-o-quinone.

    abstract::Reaction of lysine with estrone 3,4-o-quinone gave a complex mixture of products. Six compounds were isolated and identified using spectroscopic techniques. Among the reaction products isolated were 4-hydroxyestrone (2), 3-aminoisoestrone (3), 3-(N-pentyl-5-amino)-isoestrone (4), 1-lysylestrone 3,4-o-iminoquinone (5),...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Tabakovic K,Abul-Hajj YJ

    更新日期:1994-09-01 00:00:00

  • Enhancement of 2'-deoxyguanosine hydroxylation and DNA damage by coal and oil fly ash in relation to particulate metal content and availability.

    abstract::Epidemiologic studies have shown causal relationships between air pollution particles and adverse health effects in susceptible subpopulations. Fly ash particles (containing water-soluble and insoluble metals) are a component of ambient air particulate pollution and may contribute to particulate-induced health effects...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Prahalad AK,Inmon J,Ghio AJ,Gallagher JE

    更新日期:2000-10-01 00:00:00

  • Free energy profiles of base flipping in intercalative polycyclic aromatic hydrocarbon-damaged DNA duplexes: energetic and structural relationships to nucleotide excision repair susceptibility.

    abstract::The crystal structure of Rad4/Rad23, the yeast homolog of the human nucleotide excision repair (NER) lesion recognition factor XPC-RAD23B ( Min , J. H. and Pavletich , N. P. ( 2007 ) Nature 449 , 570 - 575 ) reveals that the lesion-partner base is flipped out of the helix and binds to amino acids of the protein. This ...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Cai Y,Zheng H,Ding S,Kropachev K,Schwaid AG,Tang Y,Mu H,Wang S,Geacintov NE,Zhang Y,Broyde S

    更新日期:2013-07-15 00:00:00

  • Notopterygium forbesii boiss extract and its active constituents increase reactive species and heme oxygenase-1 in human fetal hepatocytes: mechanisms of action.

    abstract::Notopterygium forbesii Boiss (NF) has been used as a traditional Chinese medicine for the treatment of common cold and rheumatism. However, there has been limited research on the biological properties of NF, and the mechanisms of action remain unknown. Here, we aimed to study the mechanism of NF-induced heme oxygenase...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Tang SY,Wang H,Zhang W,Halliwell B

    更新日期:2008-12-01 00:00:00

  • Human serum albumin-benzo[a]pyrene anti-diol epoxide adduct structure elucidation by fluorescence line narrowing spectroscopy.

    abstract::Cryogenic (4-10 K) laser-induced vibrationless ground state and vibronic excited state fluorescence emission spectra of the adducts resulting from reaction in vitro of human serum albumin and the carcinogen (+-)-r-7,t-8-dihydroxy-c-9,c-10-epoxy-7,8,9,10- tetrahydrobenzo[a]-pyrene were recorded in order to determine th...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Day BW,Doxtader MM,Rich RH,Skipper PL,Singh K,Dasari RR,Tannenbaum SR

    更新日期:1992-01-01 00:00:00

  • Inhibition of doxorubicin chronic toxicity in catalase-overexpressing transgenic mouse hearts.

    abstract::Catalase is an important antioxidant enzyme, which has been shown to provide cardiac protection from acute toxicity induced by doxorubicin, a most effective anticancer agent. Because cumulative dose-dependent chronic cardiomyopathy due to a long-term administration of doxorubicin is a significant clinical problem, the...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Kang YJ,Sun X,Chen Y,Zhou Z

    更新日期:2002-01-01 00:00:00

  • Analysis of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB)-releasing DNA adducts in human exfoliated oral mucosa cells by liquid chromatography-electrospray ionization-tandem mass spectrometry.

    abstract::Quantitation of DNA adducts could provide critical information on the relationship between exposure to tobacco smoke and cancer risk in smokers. In this study, we developed a robust and sensitive liquid chromatography-tandem mass spectrometry method for the analysis of 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB)-releasin...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Stepanov I,Muzic J,Le CT,Sebero E,Villalta P,Ma B,Jensen J,Hatsukami D,Hecht SS

    更新日期:2013-01-18 00:00:00

  • Discovery of carboxyethylpyrroles (CEPs): critical insights into AMD, autism, cancer, and wound healing from basic research on the chemistry of oxidized phospholipids.

    abstract::Basic research, exploring the hypothesis that 2-(ω-carboxyethyl)pyrrole (CEP) modifications of proteins are generated nonenzymatically in vivo is delivering a bonanza of molecular mechanistic insights into age-related macular degeneration, autism, cancer, and wound healing. CEPs are produced through covalent modificat...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章,评审


    authors: Salomon RG,Hong L,Hollyfield JG

    更新日期:2011-11-21 00:00:00

  • Mapping Chemical Respiratory Sensitization: How Useful Are Our Current Computational Tools?

    abstract::Chemical respiratory sensitization is an immunological process that manifests clinically mostly as occupational asthma and is responsible for 1 in 6 cases of adult asthma, although this may be an underestimate of the prevalence, as it is under-diagnosed. Occupational asthma results in unemployment for roughly one-thir...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Golden E,Maertens M,Hartung T,Maertens A

    更新日期:2020-12-15 00:00:00

  • Nanomaterials and Innate Immunity: A Perspective of the Current Status in Nanosafety.

    abstract::Human exposure to engineered nanomaterials (ENMs) is inevitable due to the plethora of applications for which they are being manufactured and integrated within. ENMs demonstrate plentiful advantages in terms of industrial approaches as well as from a consumer perspective. However, despite such positives, doubts remain...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Cronin JG,Jones N,Thornton CA,Jenkins GJS,Doak SH,Clift MJD

    更新日期:2020-05-18 00:00:00

  • Guanine-adenine DNA cross-linking by 1,2,3,4-diepoxybutane: potential basis for biological activity.

    abstract::1,2,3,4-Diepoxybutane (DEB) is a prominent carcinogenic metabolite of 1,3-butadiene (1,3-BD), an important industrial chemical and an environmental pollutant found in cigarette smoke and automobile exhaust. DEB is capable of inducing a variety of genotoxic effects, including point mutations, large deletions, and chrom...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Park S,Hodge J,Anderson C,Tretyakova N

    更新日期:2004-12-01 00:00:00

  • Relative toxicities of DNA cross-links and monoadducts: new insights from studies of decarbamoyl mitomycin C and mitomycin C.

    abstract::Mitomycin C (MC), a cytotoxic anticancer drug and bifunctional DNA DNA alkylating agent, induces cross-linking of the complementary strands of DNA. The DNA interstrand cross-links (ICLs) are thought to be the critical cytotoxic lesions produced by MC. Decarbamoyl mitomycin C (DMC) has been regarded as a monofunctional...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Palom Y,Suresh Kumar G,Tang LQ,Paz MM,Musser SM,Rockwell S,Tomasz M

    更新日期:2002-11-01 00:00:00

  • Molecular cytotoxicity mechanisms of allyl alcohol (acrolein) in budding yeast.

    abstract::Allyl alcohol (AA) is one of the environmental pollutants used as a herbicide and industrial chemical. AA undergoes enzymatic oxidation in vivo to form Acrolein (Acr), a highly reactive and ubiquitous environmental toxicant. The exposure to AA/Acr has detrimental effects on cells and is highly fatal. In corroboration ...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Golla U,Bandi G,Tomar RS

    更新日期:2015-06-15 00:00:00

  • Complexation of arsenic species in rabbit erythrocytes.

    abstract::The binding of arsenite, As(III), and arsenate, As(V), by molecules in the intracellular compartment of rabbit erythrocytes has been studied by 1H- and 31P-NMR spectroscopy, uptake of 73As, and ultrafiltration experiments. For intact erythrocytes to which 0.1-0.4 mM arsenite was added, direct evidence was obtained for...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Delnomdedieu M,Basti MM,Styblo M,Otvos JD,Thomas DJ

    更新日期:1994-09-01 00:00:00

  • Nontoxic and neuroprotective β-naphthotacrines for Alzheimer's disease.

    abstract::The synthesis, toxicity, neuroprotection, and human acetylcholinesterase (hAChE)/ human butyrylcholinesterase (hBuChE) inhibition properties of β-naphthotacrines1-14 as new drugs for Alzheimer's disease (AD) potential treatment, are reported. β-Naphthotacrines1-14 showed lower toxicity than tacrine; moreover, at the h...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Esquivias-Pérez M,Maalej E,Romero A,Chabchoub F,Samadi A,Marco-Contelles J,Oset-Gasque MJ

    更新日期:2013-06-17 00:00:00

  • Patterns of resistance to exonuclease digestion of oligonucleotides containing polycyclic aromatic hydrocarbon diol epoxide adducts at N6 of deoxyadenosine.

    abstract::The effect of adduct stereochemistry on the susceptibility to hydrolysis by snake venom (VPD) and bovine spleen (SPD) phosphodiesterases was investigated with short deoxyoligonucleotides containing defined adducts derived from alkylation of the exocyclic 6-amino group of dA by polycyclic aromatic hydrocarbon diol epox...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Ilankumaran P,Pannell LK,Gebreselassie P,Pilcher AS,Yagi H,Sayer JM,Jerina DM

    更新日期:2001-09-01 00:00:00

  • Gas Chromatography-Tandem Mass Spectrometry Verification of Sulfur Mustard Exposure in Humans through the Conversion of Protein Adducts to Free Sulfur Mustard.

    abstract::Exposures to sulfur mustard (HD; bis(2-chloroethyll) sulfide) are well-known to result in the formation of adducts with free aspartate and glutamate residues of plasma proteins (Lawrence, R. J., Smith, J. R., and Capacio, B. R. 2008 32, (1), 31-36). A modified version of the analytical method reported previously for t...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Young SA,Capacio BR

    更新日期:2020-07-20 00:00:00

  • Enzyme-mediated dialdehyde formation: an alternative pathway for benzo[a]pyrene 7,8-dihydrodiol bioactivation.

    abstract::Polycyclic aromatic hydrocarbons, such as benzo[a]pyrene, are widespread environmental carcinogens of human concern. Several enzymatic systems have been shown to activate benzo[a]pyrene 7, 8-dihydrodiol, the proximate carcinogenic metabolite of benzo[a]pyrene, to a reactive species which produces both a chemiluminesce...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Stansbury KH,Noll DM,Groopman JD,Trush MA

    更新日期:2000-11-01 00:00:00

  • Mechanisms of nitrogen oxide-mediated disruption of metalloprotein function: an examination of the copper-responsive yeast transcription factor Ace1.

    abstract::Nitric oxide (NO) has been found to inhibit the copper-responsive yeast transcription factor Ace1 in an oxygen-dependent manner. However, the mechanism responsible for NO-dependent inhibition of Ace1 remains unestablished. In the present study, the chemical interaction of nitrogen oxide species with Ace1 was examined ...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Shinyashiki M,Pan CJ,Switzer CH,Fukuto JM

    更新日期:2001-12-01 00:00:00

  • Binding of nickel(II) and copper(II) to the N-terminal sequence of human protamine HP2.

    abstract::A potentiometric and spectroscopic (UV/vis and CD) study of Cu(II) and Ni(II) binding to the N-terminal pentadecapeptide of human protamine HP2 (HP2(1-15)) was performed. The results indicate that the N-terminal tripeptide motif Arg-Thr-His is the exclusive binding site for both metal ions at a metal to HP2(1-15) mola...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Bal W,Jezowska-Bojczuk M,Kasprzak KS

    更新日期:1997-08-01 00:00:00

  • Inhibition of skeletal sarcoplasmic reticulum Ca2+-ATPase activity by deferoxamine nitroxide free radical.

    abstract::Deferoxamine is an inhibitor of iron-dependent free radical reactions. Despite the antioxidant roles, prolonged clinical use of the chelator is far from benign, and paradoxically, deferoxamine has been shown to promote lipid peroxidation. The possible toxicity of the drug's metabolites, such as deferoxamine nitroxide ...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章


    authors: Kiyose M,Lee CI,Okabe E

    更新日期:1999-02-01 00:00:00

  • T-Cell Activation by Low Molecular Weight Drugs and Factors That Influence Susceptibility to Drug Hypersensitivity.

    abstract::Drug hypersensitivity reactions adversely affect treatment outcome, increase the length of patients' hospitalization, and limit the prescription options available to physicians. In addition, late stage drug attrition and the withdrawal of licensed drugs cost the pharmaceutical industry billions of dollars. This signif...

    journal_title:Chemical research in toxicology

    pub_type: 杂志文章,评审


    authors: Hammond S,Thomson PJ,Ogese MO,Naisbitt DJ

    更新日期:2020-01-21 00:00:00