{rfName}
Va

Indexed in

License and Use

Icono OpenAccess

Altmetrics

Analysis of institutional authors

Perramón MAuthorParra-Robert MAuthorMorales-Ruiz MAuthorJimenez WAuthorFernández-Varo GCorresponding AuthorCasals GCorresponding Author

Share

May 6, 2021
Publications
>
Article

Validation of a Gas Chromatography-Mass Spectrometry Method for the Measurement of the Redox State Metabolic Ratios Lactate/Pyruvate and β-Hydroxybutyrate/Acetoacetate in Biological Samples

Publicated to: International Journal Of Molecular Sciences. 22 (9): 4752- - 2021-05-01 22(9), DOI: 10.3390/ijms22094752

Authors: Wijngaard, Robin; Perramon, Meritxell; Parra-Robert, Marina; Hidalgo, Susana; Butrico, Gina; Morales-Ruiz, Manuel; Zeng, Muling; Casals, Eudald; Jimenez, Wladimiro; Fernandez-Varo, Guillermo; Shulman, Gerald I; Cline, Gary W; Casals, Gregori

Affiliations

‎ Hosp Clin Univ, Ctr Invest Biomed Red Enfermedades Hepat & Digest, Inst Invest Biomed August Pi i Sunyer IDIBAPS, Serv Biochem & Mol Genet, Carrer Villarroel 170, Barcelona 08036, Spain - Author
‎ Univ Barcelona, Dept Biomed, Barcelona 08036, Spain - Author
‎ Working Grp Biochem Assessment Hepat Dis SEQCML, Barcelona 08036, Spain - Author
‎ Wuyi Univ, Sch Biotechnol & Hlth Sci, 99 Yingbing Middle Rd, Jiangmen 529020, Peoples R China - Author
‎ Yale Sch Med, Dept Internal Med, New Haven, CT 06510 USA - Author
See more

Abstract

The metabolic ratios lactate/pyruvate and beta-hydroxybutyrate/acetoacetate are considered valuable tools to evaluate the in vivo redox cellular state by estimating the free NAD+/NADH in cytoplasm and mitochondria, respectively. The aim of the current study was to validate a gas-chromatography mass spectrometry method for simultaneous determination of the four metabolites in plasma and liver tissue. The procedure included an o-phenylenediamine microwave-assisted derivatization, followed by liquid-liquid extraction with ethyl acetate and silylation with bis(trimethylsilyl)trifluoroacetamide:trimethylchlorosilane 99:1. The calibration curves presented acceptable linearity, with a limit of quantification of 0.001 mM for pyruvate, beta-hydroxybutyrate and acetoacetate and of 0.01 mM for lactate. The intra-day and inter-day accuracy and precision were within the European Medicines Agency's Guideline specifications. No significant differences were observed in the slope coefficient of three-point standard metabolite-spiked curves in plasma or liver and water, and acceptable recoveries were obtained in the metabolite-spiked samples. Applicability of the method was tested in precision-cut liver rat slices and also in HepG2 cells incubated under different experimental conditions challenging the redox state. In conclusion, the validated method presented good sensitivity, specificity and reproducibility in the quantification of lactate/pyruvate and beta-hydroxybutyrate/acetate metabolites and may be useful in the evaluation of in vivo redox states.

Keywords

acetoacetatebloodgc-msketone bodieslactatemicrowave-assisted derivatizationnicotinamide adenine dinucleotidepitfallspyruvateAcetoacetateBloodGc-msKeto-acidsKetone bodiesLactateMicrowave-assisted derivatizationNicotinamide adenine dinucleotidePitfallsPyruvateRedox state

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal International Journal Of Molecular Sciences due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2021, it was in position 69/297, thus managing to position itself as a Q1 (Primer Cuartil), in the category Biochemistry & Molecular Biology.

Independientemente del impacto esperado determinado por el canal de difusión, es importante destacar el impacto real observado de la propia aportación.

Según las diferentes agencias de indexación, el número de citas acumuladas por esta publicación hasta la fecha 2025-12-08:

  • WoS: 9
  • Scopus: 9
  • Europe PMC: 2

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-12-08:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 23.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 23 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

    It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

    • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

    Leadership analysis of institutional authors

    This work has been carried out with international collaboration, specifically with researchers from: China; United States of America.

    There is a significant leadership presence as some of the institution’s authors appear as the first or last signer, detailed as follows: First Author (Wijngaard R) and Last Author (Casals Mercadal, Gregori).

    the authors responsible for correspondence tasks have been Fernandez Varo, Guillermo and Casals Mercadal, Gregori.

    Awards linked to the item

    This study has been funded by Instituto de Salud Carlos III through the project PI19/00774 to G.C. and G.F-V, and Direccion General de Investigacion Cientifica y Tecnica RTI2018-094734-B-C2 to W.J, co-funded by European Regional Development Fund, European Union, A way to make Europe; National Natural Science Foundation of China (31950410536 to E.C.), the Wuyi University Funding (2018TP010 to E.C.), Guangdong Science and Technology Department (2019A050512006 to E.C.).