Skip to main content

Advertisement

SpringerLink
Log in

Isoprenoid quantitation in human brain tissue: a validated HPLC–fluorescence detection method for endogenous farnesyl- (FPP) and geranylgeranylpyrophosphate (GGPP)

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Farnesyl- and geranylgeranylpyrophosphate (FPP and GGPP) are isoprenoid intermediates in the mevalonate pathway. They play a crucial role in cell survival, growth and differentiation due to their attachment (isoprenylation) to small GTPases (Ras, Rho, etc.). Isoprenoid formation seems to be tightly regulated within the mevalonate pathway and its perturbation has been linked to certain diseases (e.g., cancer, Alzheimer’s disease), but tissue levels are unknown. It is therefore of the utmost importance to quantify these isoprenoids in diseased tissue or in tissue after drug administration. The current work describes an isolation procedure utilizing a combination of Extrelut® liquid/liquid and reversed-phase solid-phase extraction (SPE) for homogenized human frontal cortex tissue. In addition, after a careful validation of an HPLC–fluorescence method, this assay allowed the determination of nanomolar concentrations of endogenous FPP and GGPP levels (4.5 and 10.6 ng/mg protein, respectively) in human brain tissue. The method is selective, precise (<15% RSD), accurate (<15% relative error) and sensitive over a linear range of 10–400 ng/mL for FPP and 50–1000 ng/mL for GGPP according to the current FDA criteria for bioanalytical method validation. Overall, this new method introduces the ability to simultaneously quantify FPP and GGPP in human brain tissue, and is potentially applicable to several other tissues and species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  1. McTaggart SJ (2006) Cell Mol Life Sci 63:255–267

    Article  CAS  Google Scholar 

  2. Calleros L, Lasa M, Rodriguez-Alvarez FJ, Toro MJ, Chiloeches A (2006) Apoptosis 11:1161–1173

    Article  CAS  Google Scholar 

  3. Liao JK (2002) J Clin Invest 110:285–288

    CAS  Google Scholar 

  4. Buhaescu I, Izzedine H (2007) Clin Biochem 40:575–584

    Article  CAS  Google Scholar 

  5. Cole SL, Vassar R (2006) Neurobiol Dis 22:209–222

    Article  CAS  Google Scholar 

  6. Motoshima H, Goldstein BJ, Igata M, Araki EJ (2006) Physiol 574:63–71

    Article  CAS  Google Scholar 

  7. Duncan RE, El-Sohemy A, Archer MC (2005) Mol Nutr Food Res 49:93–100

    Article  CAS  Google Scholar 

  8. Brunner TB, Hahn SM, Gupta AK, Muschel RJ, McKenna WG, Bernhard EJ (2003) Cancer Res 63:5656–5668

    CAS  Google Scholar 

  9. Barnidge DR, Dratz EA, Martin T, Bonilla LE, Moran LB, Lindall A (2003) Anal Chem 75:445–451

    Article  CAS  Google Scholar 

  10. Downard J (2003) Nat Rev Cancer 3:11–22

    Article  Google Scholar 

  11. Sahai E, Marshall CJ (2003) Nat Rev Cancer 2:133–142

    Article  Google Scholar 

  12. Farooqui AA, Ong WY, Horrocks LA, Chen P, Farooqui T (2007) Brain Res Rev 56:443–471

    Article  CAS  Google Scholar 

  13. Eckert GP, Müller WE, Wood WG (2007) Future Lipidol 2:423–432

    Article  CAS  Google Scholar 

  14. Bruenger E, Rilling HC (1988) Anal Biochem 173:321–327

    Article  CAS  Google Scholar 

  15. Saisho Y, Morimoto A, Umeda T (1997) Anal Biochem 252:89–95

    Article  CAS  Google Scholar 

  16. Tong H, Holstein SA, Hohl RJ (2005) Anal Biochem 336:51–59

    Article  CAS  Google Scholar 

  17. Tong H, Wiemer AJ, Neighbors JD, Hohl RJ (2008) Anal Biochem 378(2):138–143

    Google Scholar 

  18. Adair WL, Cafmeyer N, Keller RK (1984) J Biol Chem 7:4441–4446

    Google Scholar 

  19. CDER (2001) Guidance for industry: bioanalytical method validation. Center for Drug Evaluation and Research (CDER), US Food and Drug Administration, Rockville, MD (see http://www.fda.gov/cder/guidance/4252fnl.pdf, last accessed 25 July 2008)

  20. Naasner M, Mergler M, Wolf K, Schuphan I (2001) J Chromatogr A 945:133–138

    Article  Google Scholar 

  21. Appels NM, Rosing H, Stephens TC, Schellens JH, Beijnen JH (2006) Anal Chem 78:2617–2622

    Article  CAS  Google Scholar 

  22. Cordle A, Koenigsknecht-Talboo J, Wilkinson B, Limpert A, Landreth GJ (2005) Biol Chem 280:34202–32409

    Article  CAS  Google Scholar 

  23. Dursina B, Reents R, Delon C, Wu Y, Kulharia M, Thutewohl M, Veligodsky A, Kalinin A, Evstifeev V, Ciobanu D, Szedlacsek SE, Waldmann H, Goody RS, Alexandrov K (2006) J Am Chem Soc 128:2822–2835

    Article  CAS  Google Scholar 

  24. Jemal M, Ouyang Z, Powell ML (2002) Rapid Commun Mass Spectrom 16:1538–1547

    Article  CAS  Google Scholar 

  25. Viswanathan CT, Bansal S, Booth B, DeStefano AJ, Rose MJ, Sailstad J, Shah VP, Skelly JP, Swann PG, Viswanathan RW (2007) Pharm Res 24:1962–1973

    Article  CAS  Google Scholar 

  26. Jemal M, Schuster A, Whigan DB (2003) Rapid Commun Mass Spectrom 17:1723–1734

    Article  CAS  Google Scholar 

  27. Li W, Cohen LH (2003) Anal Chem 75:5854–5859

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank the Hanna Bragard-Apfel Foundation, the L.A. Brain Bank, and the Medical Research Council, as well as to acknowledge the provision of NIH grants AG23524, AG18357 and the support of the Department of Veterans Affairs. We also appreciate the technical support of Rambod Daneshfar.

Author information

Authors and Affiliations

  1. Department of Pharmacology, Biocenter Niederursel, University of Frankfurt, Max-von-Laue-Str. 9, 60438, Frankfurt, Germany

    Gero P. Hooff, Walter E. Müller & Gunter P. Eckert

  2. Department of Pharmacology, University of Minnesota School of Medicine, Geriatric Research, Education and Clinical Center, VA Medical Center, Minneapolis, MN, 55417, USA

    W. Gibson Wood

  3. Medical Research Council, Human Nutrition Research, Elsie Widdowson Laboratory, Fulbourn Road, Cambridge, CB1 9NL, UK

    Dietrich A. Volmer

Authors
  1. Gero P. Hooff
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dietrich A. Volmer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. W. Gibson Wood
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Walter E. Müller
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gunter P. Eckert
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dietrich A. Volmer.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hooff, G.P., Volmer, D.A., Wood, W.G. et al. Isoprenoid quantitation in human brain tissue: a validated HPLC–fluorescence detection method for endogenous farnesyl- (FPP) and geranylgeranylpyrophosphate (GGPP). Anal Bioanal Chem 392, 673–680 (2008). https://doi.org/10.1007/s00216-008-2306-3

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-008-2306-3

Keywords

Search

Navigation