A Fast and Accurate Way to Determine Short Chain Fatty Acids in Human Serum by GC–MS and Their Distribution in Children with Digestive Diseases

  • Rui Wang
  • Chaonan Fan
  • Xiuqin Fan
  • Yunfeng Zhao
  • Yuanyuan Wang
  • Ping Li
  • Tiantian Tang
  • Hongyang Yao
  • Si Chen
  • Dawei ChenEmail author
  • Kemin QiEmail author


Short-Chain Fatty Acids (SCFAs) represent the largest group of metabolic nutrients produced by gut microbiota, and can be considered potential biomarkers for digestive tract diseases. A fast, cost-effective, and reproducible analytical method for the analysis of SCFAs is highly required. This work developed a one-step extraction of 14 SCFAs using a mixture of ethanol and n-hexane in human serum samples by GC–MS without derivatization. The recoveries of the 14 SCFAs ranged from 62.4% to 129.2% in human serum samples. The LODs and LOQs of the 14 SCFAs were 0.12–0.48 mg/L and 0.40–1.61 mg/L, respectively. The application of this method was carried out by comparing SCFAs in serum samples from children with health and digestive tract diseases, and the results demonstrated that SCFAs in the serum of children with digestive disorders were significantly higher than those of healthy children, implying that digestive tract diseases affect gut microbiota and their metabolisms.

Graphic abstract


Gas chromatography–mass spectrometry Short-chain fatty acids (SCFAs) Human serum Digestive tract diseases 



This work is financially supported by the National Natural Science Foundation of China (No. 81803214). We express our thanks to Prof. Bing Shao and Prof. Jing Zhang from Beijing CDC providing the serum samples of healthy children.


This study was funded by the National Natural Science Foundation of China (Grant number: No. 81803214).

Compliance with Ethical Standards

Conflict of interest

All authors have been informed of the full content and approved the publication of this manuscript, and all authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in the study involving human participants were in accordance with the ethical standards of Beijing Children’s Hospital and Beijing CDC research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards (case number: 2016-10).

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10337_2019_3831_MOESM1_ESM.docx (817 kb)
Supplementary material 1 (DOCX 816 kb)


  1. 1.
    Han J, Lin K, Sequeira C, Borchers CH (2015) An isotope-labeled chemical derivatization method for the quantitation of short-chain fatty acids in human feces by liquid chromatography–tandem mass spectrometry. Anal Chim Acta 854:86–94CrossRefGoogle Scholar
  2. 2.
    Primec M, Mičetić-Turk D, Langerholc T (2017) Analysis of short-chain fatty acids in human feces: a scoping review. Anal Biochem 526:9–21CrossRefGoogle Scholar
  3. 3.
    Koh A, De Vadder F, Kovatcheva-Datchary P, Backhed F (2016) From dietary fiber to host physiology: short-chain fatty acids as key bacterial metabolites. Cell 165:1332–1345CrossRefGoogle Scholar
  4. 4.
    Jiang L, Hu X, Yin D, Zhang H, Yu Z (2011) Occurrence, distribution and seasonal variation of antibiotics in the Huangpu River, Shanghai, China. Chemosphere 82:822–828CrossRefGoogle Scholar
  5. 5.
    Byrne CS, Chambers ES, Morrison DJ, Frost G (2015) The role of short chain fatty acids in appetite regulation and energy homeostasis. Int J Obes 39:1331–1338CrossRefGoogle Scholar
  6. 6.
    Tan J, McKenzie C, Potamitis M, Thorburn AN, Mackay CR, Macia L (2014) The role of short-chain fatty acids in health and disease. Adv Immunol 121:91–119CrossRefGoogle Scholar
  7. 7.
    Chambers ES, Morrison DJ, Frost G (2015) Control of appetite and energy intake by SCFA: what are the potential underlying mechanisms? Proc Nutr Soc 74:328–336CrossRefGoogle Scholar
  8. 8.
    Li X, Shimizu Y, Kimura I (2017) Gut microbial metabolite short-chain fatty acids and obesity. Biosci Microbiota Food Health 36:135–140CrossRefGoogle Scholar
  9. 9.
    Bajaj JS, Barbara G, DuPont HL, Mearin F, Gasbarrini A, Tack J (2018) New concepts on intestinal microbiota and the role of the non-absorbable antibiotics with special reference to rifaximin in digestive diseases. Dig Liver Dis 50:741–749CrossRefGoogle Scholar
  10. 10.
    Pouteau E, Nguyen P, Ballevre O, Krempf M (2003) Production rates and metabolism of short-chain fatty acids in the colon and whole body using stable isotopes. Proc Nutr Soc 62:87–93CrossRefGoogle Scholar
  11. 11.
    Zhang C, Tang P, Xu H, Weng Y, Tang Q, Zhao H (2018) Analysis of short-chain fatty acids in fecal samples by headspace gas chromatography. Chromatographia 81:1317–1323CrossRefGoogle Scholar
  12. 12.
    Zhao R, Chu L, Wang Y, Song Y, Liu P, Li C, Huang J, Kang X (2017) Application of packed-fiber solid-phase extraction coupled with GC–MS for the determination of short-chain fatty acids in children’s urine. Clin Chim Acta 468:120–125CrossRefGoogle Scholar
  13. 13.
    Fiori J, Turroni S, Candela M, Brigidi P, Gotti R (2018) Simultaneous HS–SPME GC–MS determination of short chain fatty acids, trimethylamine and trimethylamine N-oxide for gut microbiota metabolic profile. Talanta 189:573–578CrossRefGoogle Scholar
  14. 14.
    Han X, Guo J, You Y, Yin M, Ren C, Zhan J, Huang W (2018) A fast and accurate way to determine short chain fatty acids in mouse feces based on GC–MS. J Chromatogr B 1099:73–82CrossRefGoogle Scholar
  15. 15.
    Banel A, Jakimska A, Wasielewska M, Wolska L, Zygmunt B (2012) Determination of SCFAs in water using GC-FID: selection of the separation system. Anal Chim Acta 716:24–27CrossRefGoogle Scholar
  16. 16.
    Park NH, Kim MS, Lee W, Leea ME, Hong J (2017) An in situ extraction and derivatization method for rapid analysis of short-chain fatty acids in rat fecal samples by gas chromatography tandem mass spectrometry. Anal Methods 9:2351–2356CrossRefGoogle Scholar
  17. 17.
    Zhang S, Wang H, Zhu M-J (2019) A sensitive GC/MS detection method for analyzing microbial metabolites short chain fatty acids in fecal and serum samples. Talanta 196:249–254CrossRefGoogle Scholar
  18. 18.
    Furuhashi T, Sugitate K, Nakai T, Jikumaru Y, Ishihara G (2018) Rapid profiling method for mammalian feces short chain fatty acids by GC–MS. Anal Biochem 543:51–54CrossRefGoogle Scholar
  19. 19.
    Miwa H, Hiyama C, Yamamoto M (1985) High-performance liquid chromatography of short-and long-chain fatty acids as 2-nitrophenylhydrazides. J Chromatogr A 321:165–174CrossRefGoogle Scholar
  20. 20.
    Schiffels J, Baumann MEM, Selmer T (2011) Facile analysis of short-chain fatty acids as 4-nitrophenyl esters in complex anaerobic fermentation samples by high performance liquid chromatography. J Chromatogr A 1218:5848–5851CrossRefGoogle Scholar
  21. 21.
    Zheng J, Zheng S-J, Cai W-J, Yu L, Yuan B-F, Feng Y-Q (2019) Stable isotope labeling combined with liquid chromatography-tandem mass spectrometry for comprehensive analysis of short-chain fatty acids. Anal Chim Acta 1070:51–59CrossRefGoogle Scholar
  22. 22.
    Chan JCY, Kioh DYQ, Yap GC, Lee BW, Chan ECY (2017) A novel LCMSMS method for quantitative measurement of short-chain fatty acids in human stool derivatized with 12C- and 13C-labelled aniline. J Pharm Biomed Anal 138:43–53CrossRefGoogle Scholar
  23. 23.
    Jakobsdottir G, Bjerregaard JH, Skovbjerg H, Nyman M (2013) Fasting serum concentration of short-chain fatty acids in subjects with microscopic colitis and celiac disease: no difference compared with controls, but between genders. Scand J Gastroenterol 48:696–701CrossRefGoogle Scholar
  24. 24.
    Bruce SJ, Tavazzi I, Parisod V, Rezzi S, Kochhar S, Guy PA (2009) Investigation of human blood plasma sample preparation for performing metabolomics using ultrahigh performance liquid chromatography/mass spectrometry. Anal Chem 81:3285–3296CrossRefGoogle Scholar
  25. 25.
    Garcia-Villalba R, Gimenez-Bastida JA, Garcia-Conesa MT, Tomas-Barberan FA, Carlos Espin J, Larrosa M (2012) Alternative method for gas chromatography-mass spectrometry analysis of short-chain fatty acids in faecal samples. J Sep Sci 35:1906–1913CrossRefGoogle Scholar
  26. 26.
    Escorsim AM, da Rocha G, Vargas JVC, Mariano AB, Ramos LP, Corazza ML, Cordeiro CS (2018) Extraction of Acutodesmus obliquus lipids using a mixture of ethanol and hexane as solvent. Biomass Bioenergy 108:470–478CrossRefGoogle Scholar
  27. 27.
    Zeng M, Cao H (2018) Fast quantification of short chain fatty acids and ketone bodies by liquid chromatography–tandem mass spectrometry after facile derivatization coupled with liquid-liquid extraction. J Chromatogr B 1083:137–145CrossRefGoogle Scholar
  28. 28.
    Lotti C, Rubert J, Fava F, Tuohy K, Mattivi F, Vrhovsek U (2017) Development of a fast and cost-effective gas chromatography–mass spectrometry method for the quantification of short-chain and medium-chain fatty acids in human biofluids. Anal Bioanal Chem 409:5555–5567CrossRefGoogle Scholar
  29. 29.
    McNabney SM, Henagan TM (2017) Short chain fatty acids in the colon and peripheral tissues: a focus on butyrate, colon cancer, obesity and insulin resistance. Nutrients 9:138CrossRefGoogle Scholar
  30. 30.
    Perry RJ, Peng L, Barry NA, Cline GW, Zhang D, Cardone RL, Petersen KF, Kibbey RG, Goodman AL, Shulman GI (2016) Acetate mediates a microbiome-brain-beta-cell axis to promote metabolic syndrome. Nature 534:213–217CrossRefGoogle Scholar
  31. 31.
    Bian X, Li N, Tan B, Sun B, Guo MQ, Huang G, Fu L, Wendy Hsiao WL, Liu L, Wu JL (2018) Polarity-tuning derivatization-LC–MS approach for probing global carboxyl-containing metabolites in colorectal cancer. Anal Chem 90:11210–11215CrossRefGoogle Scholar
  32. 32.
    Wong C, Harris PJ, Ferguson LR (2016) Potential benefits of dietary fibre intervention in inflammatory bowel disease. Int J Mol Sci 17CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Rui Wang
    • 1
  • Chaonan Fan
    • 1
  • Xiuqin Fan
    • 1
  • Yunfeng Zhao
    • 2
    • 3
  • Yuanyuan Wang
    • 1
  • Ping Li
    • 1
  • Tiantian Tang
    • 1
  • Hongyang Yao
    • 1
  • Si Chen
    • 1
  • Dawei Chen
    • 2
    • 3
    Email author
  • Kemin Qi
    • 1
    Email author
  1. 1.Laboratory of Nutrition and Development, Beijing Pediatric Research Institute, Beijing Children’s HospitalCapital Medical University, National Center for Children’s HealthBeijingPeople’s Republic of China
  2. 2.China National Center for Food Safety Risk AssessmentBeijingPeople’s Republic of China
  3. 3.Key Laboratory of Food Safety Risk AssessmentMinistry of HealthBeijingPeople’s Republic of China

Personalised recommendations