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Fast Determination of Short-Chain Fatty Acids and Glucose Simultaneously by Ultraviolet/Visible and Refraction Index Detectors via High-Performance Liquid Chromatography

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Abstract

A rapid analytical method for simultaneous quantification of glucose (GLC) and seven short-chain fatty acids (SCFA) was developed for detection of ruminal fermentation parameters. The formic acid (FA), lactic acid (LA), acetic acid (AA), propionic acid (PA), butyric acid (BA), isovaleric acid (IVA), valeric acid (VA), and glucose were identified and quantified by high-performance liquid chromatography (HPLC) technique, using a reverse-phase column (C-18) and simultaneous detection in UV-Vis and Refraction Index (RID) detectors, linked in series. The method was validated following the requirements of selectivity, linearity, sensitivity, limit of detection, limit of quantification, accuracy, and precision. The developed method operates in isocratic and gradient modes, providing a better SCFA separation, which allows their determination with a high accuracy and repeatability. Therefore, this study provides a fast, reliable, accurate, straightforward, and efficient alternative method for analysis of ruminal fermentation parameters by liquid chromatography using both detectors simultaneously.

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References

  • Agência Nacional de Vigilância Sanitária (ANVISA); Resolução RE n° 899. (accessed 18 Jan 2020)

  • Armbruster DA, Pry T (2008) Limit of blank, limit of detection and limit of quantitation. Clin Biochem Rev 29:S49–S52

    PubMed  PubMed Central  Google Scholar 

  • Baldwin RL (1998) Use of isolated ruminal epithelial cells in the study of rumen metabolism. J Nutr 128:293–296

    Article  Google Scholar 

  • Bergman EN (1990) Energy contributions of volatile fatty acids from the gastrointestinal tract in various species. Physiol Rev 70:567–590

    Article  CAS  Google Scholar 

  • Brito NM, Junior OPA, Polese L, Ribeiro ML (2003) Validação de métodos analíticos: estratégia e discussão. Pesticidas: Revista de Ecotoxicologia e Meio Ambiente 13:129–146

    CAS  Google Scholar 

  • Castellari M, Sartini E, Spinabelli U, Riponi C, Galassi S (2001) Determination of carboxylic acids, carbohydrates, glycerol, ethanol, and 5-HMF in beer by high-performance liquid chromatography and UV-refractive index double detection. J Chromatogr Sci 39:235–238

    Article  CAS  Google Scholar 

  • Cottyn BG, Boucque CV (1968) Rapid Method for the Gas-Chromatographic Determination of Volatile Fatty Acids in Rumen Fluid. J Agric Food Chem 16:105–107

    Article  CAS  Google Scholar 

  • De Baere S, Eeckhaut V, Steppe M, De Maesschalck C, De Becker P, Van Immerseel F, Croubels S (2013) Development of a HPLC–UV method for the quantitative determination of four short-chain fatty acids and lactic acid produced by intestinal bacteria during in vitro fermentation. J Pharm Biomed Anal 80:107–115

    Article  Google Scholar 

  • de Sá LRV, Oliveira MAL, Matos A, Cammarota MC, Ferreira-Leitão VS (2011) Análise simultânea de carboidratos e ácidos orgânicos voláteis por CLAE para monitoramento do processo de produção de biohidrogênio por via fermentativa. In: Embrapa Solos-Resumo em anais de congresso (ALICE). In: LATIN AMERICAN WORKSHOP AND SYMPOSIUM ON ANAEROBIC DIGESTION, Ouro Preto, Brazil, pp. 5

  • Dijkstra J (1994) Production and absorption of volatile fatty acids in the rumen. Livest Prod Sci 39:61–69

    Article  Google Scholar 

  • Filipek J, Dvorak R (2009) Determination of the Volatile Fatty Acid Content in the Rumen Liquid: Comparison of Gas Chromatography and Capillary Isotachophoresis. Acta Vet Brno 78:627–633

    Article  CAS  Google Scholar 

  • Goularte SR, Ítavo LCV, Santos GT, Ítavo CCBF, Oliveira LCS, Favaro SP, Dias AM, Torres Junior RAA, Bittas CMM (2011) Ácidos graxos voláteis no rúmen de vacas alimentadas com diferentes teores de concentrado na dieta. Arq Bras Med Vet Zootecnia 63:1479–1486

    Article  CAS  Google Scholar 

  • Guerrant GO, Lambert MA, Moss CW (1982) Analysis of short-chain acids from anaerobic bacteria by high-performance liquid chromatography. J Clin Microbiol 16:355–360

    Article  CAS  Google Scholar 

  • Huntington GB (1990) Energy metabolism in the digestive tract and liver of cattle: influence of physiological state and nutrition. Reprod Nutr Dev 30:35–47

    Article  CAS  Google Scholar 

  • Instituto Nacional de Metrologia, Normalização e Qualidade Industrial (INMETRO); Orientações sobre validação de métodos analíticos. (accessed 27 Jan 2020)

  • Kim HJ, Lee MJ, Kim HJ, Cho SK (2017) Development of HPLC-UV Method for Detection and Quantification of Eight Organic Acids in Animal Feed. J Chromatogr Sep Tech 8:1–5

    Article  Google Scholar 

  • López EF, Gómez EF (1996) Simultaneous determination of the major organic acids, sugars, glycerol, and ethanol by HPLC in grape musts and white wines. J Chromatogr Sci 34:254–257

    Article  Google Scholar 

  • Mesquita PL, Afonso RJCF, Aquino SF, Leite GS (2013) Validação de Método de cromatografia líquida para a determinação de sete ácidos graxos voláteis intermediários da digestão anaeróbia. Engenharia Sanitaria e Ambiental 18:295–302

    Article  Google Scholar 

  • Olson KC, Cochran RC, Jones TJ, Vanzant ES, Titgemeyer EC, Johnson DE (1999) Effects of ruminal administration of degradable intake protein and starch on utilization of low-quality warm season grass hay by beef steers. J Anim Sci 77:1016–1025

    Article  CAS  Google Scholar 

  • Stein J, Kulemeier J, Lembcke B, Caspary WF (1992) Simple and rapid method for determination of short-chain fatty acids in biological materials by high-performance liquid chromatography with ultraviolet detection. J Chromatogr B Biomed Sci Appl 576:53–61

    Article  CAS  Google Scholar 

  • Thompson M, Ellison SL, & Wood R (2002) Harmonized guidelines for single-laboratory validation of methods of analysis (IUPAC Technical Report). Pure Appl Chem 74(5):835-856

  • Valadares Filho SC, Pina DS (2011) Fermentação Ruminal. In: Nutrição de Ruminantes, 2nd edn. FUNEP, Jaboticabal, pp 161–189

    Google Scholar 

  • Van Soest PJ (ed) (2018) Nutritional ecology of the ruminant. Cornell University Press, Ithaca

    Google Scholar 

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Acknowledgments

The authors thank the São Paulo Research Foundation (FAPESP) by supporting this work and the São Paulo State University (Unesp), School of Sciences and Engineering, Tupã, for kindly providing all the conditions for the experimental analyses performed in this work.

Funding

This work was entirely supported by the São Paulo Research Foundation (FAPESP) by processes numbers 2016/00446-7 and 2017/10393-0. E. F. V. and J. A. S. thanks São Paulo State University (Unesp) for PROPe grant number 426 (from Edital 06/2019). This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) Finance Code 001 (to R. F. S.).

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Authors and Affiliations

  1. São Paulo State University (Unesp), School of Sciences and Engineering, Tupã, São Paulo, Brazil

    Jéssica Aparecida Serafim & Eduardo Festozo Vicente

  2. São Paulo State University (Unesp), School of Agricultural and Veterinarian Sciences, Jaboticabal, São Paulo, Brazil

    Raiza Felismino Silveira

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  1. Jéssica Aparecida Serafim
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  2. Raiza Felismino Silveira
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  3. Eduardo Festozo Vicente
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Correspondence to Eduardo Festozo Vicente.

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All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

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J. A. S. declares that she has no conflict of interest. R. F. S. declares that she has no conflict of interest. E. F. V. declares that he has no conflict of interest.

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Serafim, J.A., Silveira, R.F. & Vicente, E.F. Fast Determination of Short-Chain Fatty Acids and Glucose Simultaneously by Ultraviolet/Visible and Refraction Index Detectors via High-Performance Liquid Chromatography. Food Anal. Methods 14, 1387–1393 (2021). https://doi.org/10.1007/s12161-021-01990-w

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