Analytical and Bioanalytical Chemistry

, Volume 391, Issue 8, pp 2881–2889 | Cite as

GC-MS with ethyl chloroformate derivatization for comprehensive analysis of metabolites in serum and its application to human uremia

  • Xiumei Tao
  • Yumin Liu
  • Yihuang Wang
  • Yunping Qiu
  • Jingchao Lin
  • Aihua Zhao
  • Mingming Su
  • Wei Jia
Original Paper
First Online:
Received:
Revised:
Accepted:

Abstract

An optimized method based on GC-MS with ethyl chloroformate derivatization has been developed for the comprehensive analysis of endogenous metabolites in serum. Twenty-two reference standards and serum samples were used to validate the proposed method. The correlation coefficient was higher than 0.9900 for each of the standards, and the LOD varied from 125 to 300 pg on-column. The analytical equipment exhibited good repeatability (RSD<10%) for all of the standards. Both the repeatability and the within-48-h stability of the analytical method were satisfactory (RSD<10%) for the 18 metabolites identified in the serum samples. Mean recovery was acceptable for the 18 metabolites, ranging from 70% to 120% with RSDs of less than 10%. Using the optimized protocol and a subsequent multivariate statistical technique, complete differentiation was achieved between the metabolic profile of uremic patients and that of age- and sex-matched normal subjects. Significantly decreased levels of valine, leucine, and isoleucine and increased levels of myristic acid and linoleic acid were observed in the patient group. This work demonstrated that this method is suitable for serum-based metabolic profiling studies.

Keywords

Metabolic profiling GC-MS Ethyl chloroformate Serum Multivariate statistical analysis Metabonomics GC Bioanalytical methods Clinical/biomedical analysis 

Abbreviations

GC-MS

Gas chromatography–mass spectrometry

LOD

Limit of detection

RSD

Relative standard deviation

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Notes

Acknowledgment

This study was financially supported by the Research Projects of the Shanghai Science and Technology Commission (Project Number 06DZ19707, 064307067, 06DZ05906).

References

  1. 1.
    Calvert VS, Collantes R, Elariny H, Afendy A, Baranova A, Mendoza M, Goodman Z, Liotta LA, Petricoin EF, Younossi ZM (2007) Hepatology 46:166–172CrossRefGoogle Scholar
  2. 2.
    Andrews SS, Arkin AP (2007) Curr Biol 17:410–412CrossRefGoogle Scholar
  3. 3.
    Larive CK (2007) Anal Bioanal Chem 387:523CrossRefGoogle Scholar
  4. 4.
    Nicholson JK, Lindon JC, Holmes E (1999) Xenobiotica 29:1181–1189CrossRefGoogle Scholar
  5. 5.
    Ott KH, Aranibar N (2007) Methods Mol Biol 358:247–271CrossRefGoogle Scholar
  6. 6.
    Yap IK, Clayton TA, Tang H, Everett JR, Hanton G, Provost JP, Le Net JL, Charuel C, Lindon JC, Nicholson JK (2006) J Proteome Res 5:2675–2684CrossRefGoogle Scholar
  7. 7.
    Pan Z, Raftery D (2007) Anal Bioanal Chem 387:525–527CrossRefGoogle Scholar
  8. 8.
    Huhn SD, Szabo CM, Gass JH, Manzi AE (2004) Anal Bioanal Chem 378:1511–1519CrossRefGoogle Scholar
  9. 9.
    Williams RE, Lenz EM, Lowden JS, Rantalainen M, Wilson ID (2005) Mol Biosyst 1:166–175CrossRefGoogle Scholar
  10. 10.
    Nicholls AW, Wilson ID, Godejohann M, Nicholson JK, Shockcor JP (2006) Xenobiotica 36:615–629CrossRefGoogle Scholar
  11. 11.
    Yang J, Xu G, Zheng Y, Kong H, Wang C, Zhao X, Pang T (2005) J Chromatogr A 1084:214–212CrossRefGoogle Scholar
  12. 12.
    Gao P, Shi C, Tian J, Shi X, Yuan K, Lu X, Xu G (2007) J Pharm Biomed Anal 44:180–187CrossRefGoogle Scholar
  13. 13.
    Xuan Y, Weber G, Manz A (2006) J Chromatogr A 1130:212–218CrossRefGoogle Scholar
  14. 14.
    Jiye A, Johan T, Jonas G, Annika IJ, Par J, Henrik A, Stefan L, Marklund TM (2005) Anal Chem 77:8086–8094CrossRefGoogle Scholar
  15. 15.
    Qiu Y, Su M, Liu Y, Chen M, Gu J, Zhang J, Jia W (2007) Anal Chim Acta 583:277–283Google Scholar
  16. 16.
    Janiece LH, Bryan JP, Erik JN, Mary EL, Robert E (2005) Talenta 65:380–388CrossRefGoogle Scholar
  17. 17.
    Atsuchi S, Hiromi S, Yasuji K, Takashi F (2003) J Chromatogr B 794:373–380CrossRefGoogle Scholar
  18. 18.
    Haberhaucer TC, Alvare LG, Zitting E, Rodriguez GP, Roserberg E, Sanz MA (2003) J Chromatogr A1015:1–10Google Scholar
  19. 19.
    Husek P, Liebich HM (1994) J Chromatogr B 656:37–43CrossRefGoogle Scholar
  20. 20.
    Husek P, Matucha P, Vrankova A, Simek P (2003) J Chromatogr B 789:311–322CrossRefGoogle Scholar
  21. 21.
    Timothy WM, Thomas HH (2007) N Engl J Med 357:1316–1325CrossRefGoogle Scholar
  22. 22.
    Preethi Y, Poorna RK, Laura H, Jennifer H, Adam WC (2007) J Am Soc Hypertens 1:178–184Google Scholar
  23. 23.
    Ni Y, Su M, Qiu Y, Chen M, Liu Y, Zhao A, Jia W (2007) FEBS Lett 581:707–711CrossRefGoogle Scholar
  24. 24.
    Li H, Ni Y, Su M, Qiu Y, Zhou M, Qiu M, Zhao A, Zhao L, Jia W (2007) J Proteome Res 6:1364–1370CrossRefGoogle Scholar
  25. 25.
    Zomer S, Guillo C, Brereton RG, Hanna-Brown M (2004) Anal Bioanal Chem 378:2008–2020CrossRefGoogle Scholar
  26. 26.
    Krystyna P, Dariusz P, Michal M (2007) Clin Biochem 40:81–88CrossRefGoogle Scholar
  27. 27.
    Hide AM, Mitchell LA, Bradbury JA, Qu W, Zeldin DC, Schnellmann RG, Grant DF (2000) Toxicol Appl Pharmacol 168:268–279CrossRefGoogle Scholar
  28. 28.
    Matera M, Bellinghieri G, Costantino G, Santoro D, Calvani M, Savica V (2003) J Ren Nutr 13:2–14CrossRefGoogle Scholar
  29. 29.
    Ceballos I, Chauveau P, Guerin V, Bardet J, Parvy P, Kamoun P, Jungers P (1990) Clin Chim Acta 188:101–108Google Scholar
  30. 30.
    Alvestrand A, Furst P, Bergstrom J (1982) Clin Nephrol 18:297–305Google Scholar
  31. 31.
    Dubnov G, Berry EM (2004) Metab Syndr Relat Disord 2:124–128CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Xiumei Tao
    • 1
    • 2
  • Yumin Liu
    • 1
  • Yihuang Wang
    • 3
  • Yunping Qiu
    • 3
  • Jingchao Lin
    • 1
  • Aihua Zhao
    • 1
  • Mingming Su
    • 1
  • Wei Jia
    • 1
    • 2
  1. 1.School of PharmacyShanghai Jiao Tong UniversityShanghaiChina
  2. 2.Shanghai University of Traditional Chinese MedicineShanghaiChina
  3. 3.Shanghai Institute for Systems BiomedicineShanghai Jiao Tong UniversityShanghaiChina

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