Skip to main content
SpringerLink
Log in

Measurement of Oxidized and Reduced Coenzyme Q in Biological Fluids, Cells, and Tissues: An HPLC-EC Method

  • Protocol
  • First Online:
Mitochondrial Disorders

Part of the book series: Methods in Molecular Biology ((MIMB,volume 837))

Abstract

Direct measure of coenzyme Q (CoQ) in biological specimens may provide important advantages. Precise and selective high-performance liquid chromatography (HPLC) methods with electrochemical (EC) detection have been developed for the measurement of reduced (ubiquinol) and oxidized (ubiquinone) CoQ in biological fluids, cells, and tissues. EC detection is preferred for measurement of CoQ because of its high sensitivity. Reduced and oxidized CoQ are first extracted from biological specimens using 1-propanol. After centrifugation, the 1-propanol supernatant is directly injected into HPLC and monitored at a dual-electrode. The EC reactions occur at the electrode surface. The first electrode transforms ubiquinone into ubiquinol, and the second electrode measures the current produced by the oxidation of the hydroquinone group of ubiquinol. The methods described provide rapid, precise, and simple procedures for determination of reduced and oxidized CoQ in biological fluids, cells, and tissues. The methods have been successfully adapted to meet regulatory requirements for clinical laboratories, and have been proven reliable for analysis of clinical and research samples for clinical trials and animal studies involving large numbers of specimens.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Turunen M., Olsson J., and Dallner G. (2004) Metabolism and function of coenzyme Q. Biochim Biophys Acta 1660, 171–199.

    Article  PubMed  CAS  Google Scholar 

  2. Bentinger M., Tekle M., and Dallner G. (2010) Coenzyme Q-biosynthesis and functions. Biochem Biophys Res Commun. 21, 396, 74–79.

    Article  Google Scholar 

  3. Barshop B.A. and Gangoiti J.A. (2007) Analysis of coenzyme Q in human blood and tissues. Mitochondrion 7 Suppl, S89–93.

    Article  PubMed  CAS  Google Scholar 

  4. Quinzii C.M., López L.C., Gilkerson R.W., et al. (2010) Reactive oxygen species, oxidative stress, and cell death correlate with level of CoQ10 deficiency. FASEB J. 24, 3733–3743.

    Article  PubMed  CAS  Google Scholar 

  5. Rodríguez-Hernández A., Cordero M.D., Salviati L., et al. (2009) Coenzyme Q deficiency triggers mitochondria degradation by mitophagy. Autophagy. 5, 19–32.

    Article  PubMed  Google Scholar 

  6. Miles L., Miles M.V., Tang P.H., et al. (2005) Coenzyme Q: a potential marker for mitochondrial activity and redox status in skeletal muscle. Pediatr Neurol 32, 318–324.

    Article  PubMed  Google Scholar 

  7. Miles L., Wong B.L., Dinopoulos A., et al. (2006) Investigation of children for mitochondriopathy confirms need for strict patient selection, improved morphological criteria, and better laboratory methods. Hum Pathol 37, 173–184.

    Article  PubMed  Google Scholar 

  8. Haas R.H., Parikh S., Falk M.J., et al. (2008) The in-depth evaluation of suspected mitochondrial disease. Mol Genet Metab 94, 16–37.

    Article  PubMed  CAS  Google Scholar 

  9. Miles M.V., Miles L., Tang P.H., et al. (2008) Systematic evaluation of muscle coenzyme Q10 content in children with mitochondrial respiratory chain deficiencies. Mitochondrion 8, 170–180.

    Article  PubMed  CAS  Google Scholar 

  10. Miles M.V., Putnam P.E., Miles L., et al. (2011) Acquired coenzyme Q10 deficiency in children with recurrent food intolerance and allergies. Mitochondrion 11, 127–135.

    Article  PubMed  CAS  Google Scholar 

  11. Quinzii C.M. and Hirano M. (2010) Coenzyme Q and mitochondrial disease. Dev Disabil Res Rev 16, 183–188.

    Article  PubMed  Google Scholar 

  12. Quinzii C.M., López L.C., Von-Moltke J., et al. (2008) Respiratory chain dysfunction and oxidative stress correlate with severity of primary CoQ10 deficiency. FASEB J 22, 1874–1885.

    Article  PubMed  CAS  Google Scholar 

  13. Villalba J.M., Parrado C., Santos-Gonzalez M., et al. (2010) Therapeutic use of coenzyme Q10 and coenzyme Q10-related compounds and formulations. Expert Opin Investig Drugs 19, 535–554.

    Article  PubMed  CAS  Google Scholar 

  14. Tang P.H., Miles M.V., DeGrauw A., et al. (2001) HPLC analysis of reduced and oxidized coenzyme Q10 in human plasma. Clin Chem 47, 256–265.

    PubMed  CAS  Google Scholar 

  15. Miles M.V., Horn P., Morrison J.A., et al. (2003) Plasma coenzyme Q10 reference intervals, but not redox status, are affected by gender and race in self-reported healthy adults. Clin Chim Acta 332, 123–132.

    Article  PubMed  CAS  Google Scholar 

  16. Miles M.V., Horn P.S., Tang P.H., et al. (2004) Age-related changes in plasma coenzyme Q10 concentrations and redox state in apparently healthy children and adults. Clin Chim Acta 347, 139–144.

    Article  PubMed  CAS  Google Scholar 

  17. Niklowitz P., Andler W., and Menke T. (2006) Coenzyme Q10 concentration in plasma and blood cells: what about diurnal changes? Biofactors 28, 47–54.

    Article  PubMed  CAS  Google Scholar 

  18. Miles M.V., Tang P.H., Miles L., et al. (2008) Validation and application of an HPLC-EC method for analysis of coenzyme Q10 in blood platelets. Biomed Chromatogr 22, 1403–1408.

    Article  PubMed  CAS  Google Scholar 

  19. Tang P.H., Miles M.V., Miles L., et al. (2004) Measurement of reduced and oxidized coenzyme Q9 and coenzyme Q10 levels in mouse tissues by HPLC with coulometric detection. Clin Chim Acta 341, 173–184.

    Article  PubMed  CAS  Google Scholar 

  20. Sacconi S., Trevisson E., Salviati L., et al. (2010) Coenzyme Q10 is frequently reduced in muscle of patients with mitochondrial myopathy. Neuromuscul Disord 20, 44–48.

    Article  PubMed  Google Scholar 

  21. Tang P.H., Miles M.V., Steele P., et al. (2006) Determination of coenzyme Q10 in human breast milk by high-performance liquid chromatography. Biomed Chromatogr 20, 1336–1343.

    Article  PubMed  CAS  Google Scholar 

  22. Colomé C., Artuch R., Vilaseca M.A. et al. (2002) Ubiquinone-10 content in lymphocytes of phenylketonuric patients. Clin Biochem 35, 81–84.

    Article  PubMed  Google Scholar 

  23. Kohli R., Kirby M., Xanthakos S.A., et al. (2010) High-fructose, medium chain trans fat diet induces liver fibrosis and elevates plasma coenzyme Q9 in a novel murine model of obesity and nonalcoholic steatohepatitis. Hepatology 52, 934–944.

    Article  PubMed  CAS  Google Scholar 

  24. Isobe C., Abe T., and Terayama Y. (2010) Levels of reduced and oxidized coenzyme Q-10 and 8-hydroxy-2′-deoxyguanosine in the cerebrospinal fluid of patients with living Parkinson’s disease demonstrate that mitochondrial oxidative damage and/or oxidative DNA damage contributes to the neurodegenerative process. Neurosci Lett 469, 159–163.

    Article  PubMed  CAS  Google Scholar 

  25. Niklowitz P., Menke T., Wiesel T., et al. (2002) Coenzyme Q10 in plasma and erythrocytes: comparison of antioxidant levels in healthy probands after oral supplementation and in patients suffering from sickle cell anemia. Clin Chim Acta 326, 155–161.

    Article  PubMed  CAS  Google Scholar 

  26. Albano C.B., Muralikrishnan D., and Ebadi M. (2002) Distribution of coenzyme Q homologues in brain. Neurochem Res 27, 359–368.

    Article  PubMed  CAS  Google Scholar 

  27. Balercia G., Buldreghini E., Vignini A., et al. (2009) Coenzyme Q10 treatment in infertile men with idiopathic asthenozoospermia: a placebo-controlled, double-blind randomized trial. Fertil Steril 91, 1785–1792.

    Article  PubMed  CAS  Google Scholar 

  28. Diomedi-Camassei F., Di Giandomenico S., Santorelli F.M., et al. (2007) COQ2 nephropathy: a newly described inherited mitochondriopathy with primary renal involvement. J Am Soc Nephrol 18, 2773–2780.

    Article  PubMed  CAS  Google Scholar 

  29. Montero R., Sánchez-Alcázar J.A., Briones P., et al. (2008) Analysis of coenzyme Q10 in muscle and fibroblasts for the diagnosis of CoQ10 deficiency syndromes. Clin Biochem 41, 697–700.

    Article  PubMed  CAS  Google Scholar 

  30. Haas D., Niklowitz P., Hörster F., et al. (2009) Coenzyme Q(10) is decreased in fibroblasts of patients with methylmalonic aciduria but not in mevalonic aciduria. J Inherit Metab Dis 32, 570–575.

    Article  PubMed  CAS  Google Scholar 

  31. DiMauro S., Quinzii C.M., and Hirano M. (2007) Mutations in coenzyme Q10 biosynthetic genes. J Clin Invest 117, 587–589.

    Article  PubMed  CAS  Google Scholar 

  32. Debray F.G., Lambert M., and Mitchell G.A. (2008) Disorders of mitochondrial function. Curr Opin Pediatr 20, 471–482.

    Article  PubMed  Google Scholar 

  33. Miles M.V., Patterson B.J., Chalfonte–Evans M., et al. (2007) Ubiquinol-10 improves oxidative imbalance in children with trisomy 21. Pediatr Neurol 37, 398–403.

    Google Scholar 

  34. Montero R., Sánchez-Alcázar J.A., Briones P., et al. (2009) Coenzyme Q10 deficiency associated with a mitochondrial DNA depletion syndrome: a case report. Clin Biochem 42, 742–745.

    Article  PubMed  CAS  Google Scholar 

  35. Haas D., Niklowitz P., Hoffmann G.F., et al. (2008) Plasma and thrombocyte levels of coenzyme Q10 in children with Smith-Lemli-Opitz syndrome (SLOS) and the influence of HMG-CoA reductase inhibitors. Biofactors 32, 191–197.

    Article  PubMed  CAS  Google Scholar 

  36. Sohmiya M., Tanaka M., Tak N.W., et al. (2004) Redox status of plasma coenzyme Q10 indicates elevated systemic oxidative stress in Parkinson’s disease. J Neurol Sci 223, 161–166.

    Article  PubMed  CAS  Google Scholar 

  37. Cooper J.M., Korlipara L.V., Hart P.E., et al. (2008) Coenzyme Q10 and vitamin E deficiency in Friedreich’s ataxia: predictor of efficacy of vitamin E and coenzyme Q10 therapy. Eur J Neurol 15, 1371–1379.

    Article  PubMed  CAS  Google Scholar 

  38. Ochoa J.J., Ramirez-Tortosa M.C., Quiles J.L., et al. (2003) Oxidative stress in erythrocytes from premature and full-term infants during their first 72 h of life. Free Radic Res 3, 317–322.

    Article  Google Scholar 

  39. Roland L., Gagné A., Bélanger M.C., et al. (2010) Existence of compensatory defense mechanisms against oxidative stress and hypertension in preeclampsia. Hypertens Pregnancy 29, 21–37.

    Article  PubMed  CAS  Google Scholar 

  40. Hershey A.D., Powers S.W., Vockell A.L., et al. (2007) Coenzyme Q10 deficiency and response to supplementation in pediatric and adolescent migraine. Headache 47, 73–80.

    Article  PubMed  Google Scholar 

  41. Laguna T.A., Sontag M.K., Osberg I., et al. (2008) Decreased total serum coenzyme-Q10 concentrations: a longitudinal study in children with cystic fibrosis. J Pediatr 153, 402–407.

    Article  PubMed  CAS  Google Scholar 

  42. Mancini A., Leone E., Silvestrini A., et al. (2010) Evaluation of antioxidant systems in pituitary-adrenal axis diseases. Pituitary 13, 138–145.

    Article  PubMed  CAS  Google Scholar 

  43. Bélanger M.C., Mirault M.E., Dewailly E., et al. (2008) Environmental contaminants and redox status of coenzyme Q10 and vitamin E in Inuit from Nunavik. Metabolism 57, 927–933.

    Article  PubMed  Google Scholar 

  44. Suzuki T., Nozawa T., Sobajima M., et al. (2008) Atorvastatin-induced changes in plasma coenzyme Q10 and brain natriuretic peptide in patients with coronary artery disease. Int Heart J 49, 423–433.

    Article  PubMed  CAS  Google Scholar 

  45. Niklowitz P., Wiesel T., Andler W., and Menke T. (2007) Coenzyme Q10 concentration in the plasma of children suffering from acute lymphoblastic leukemia before and during induction treatment. Biofactors 29, 83–89.

    Article  PubMed  CAS  Google Scholar 

  46. Steele P.E., Tang P.H., DeGrauw T.J., and Miles M.V. (2004) Clinical laboratory monitoring of coenzyme Q10 use in neurologic and muscular diseases. Am J Clin Pathol 121 (Suppl 1), S113–S120.

    PubMed  Google Scholar 

  47. Takada M., Ikenoya S., Yuzuriha T., and Katayama K. (1984) Simultaneous determination of reduced and oxidized ubiquinones. Methods Enzymol 105, 147–155.

    Article  PubMed  CAS  Google Scholar 

  48. Lang J.K. and Packer L. (1987) Quantitative determination of vitamin E and oxidized and reduced coenzyme Q by high-performance liquid chromatography with in-line ultraviolet and electrochemical detection. J Chromatogr 385, 109–117.

    Article  PubMed  CAS  Google Scholar 

  49. Edlund P.O. (1988) Determination of coenzyme Q10, alpha-tocopherol and cholesterol in biological samples by coupled-column liquid chromatography with coulometric and ultraviolet detection. J Chromatogr 425, 87–97.

    Article  PubMed  CAS  Google Scholar 

  50. Okamoto T., Fukunaga Y., Ida Y., and Kishi T. (1988) Determination of reduced and total ubiquinones in biological materials by liquid chromatography with electrochemical detection. J Chromatogr 430, 11–19.

    Article  PubMed  CAS  Google Scholar 

  51. Grossi G., Bargossi A.M., Fiorella P.L., and Piazzi S. (1992) Improved high-performance liquid chromatographic method for the determination of coenzyme Q10 in plasma. J Chromatogr 593, 217–226.

    Article  PubMed  CAS  Google Scholar 

  52. Wakabayashi H., Yamato S., Nakajima M., and Shimada K. (1994) Simultaneous determination of oxidized and reduced coenzyme Q and alpha-tocopherol in biological samples by high performance liquid chromatography with platinum catalyst reduction and electrochemical detection. Biol Pharm Bull 17, 997–1002.

    Article  PubMed  CAS  Google Scholar 

  53. Finckh B., Kontush A., Commentz J., et al. (1995) Monitoring of ubiquinol-10, ubiquinone-10, carotenoids, and tocopherols in neonatal plasma microsamples using high-performance liquid chromatography with coulometric electrochemical detection. Anal Biochem 232, 210–216.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Pathology and Laboratory Medicine, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH, USA

    Peter H. Tang

  2. Division of Pathology and Laboratory Medicine, Departments of Pediatrics and Pathology & Laboratory Medicine, Cincinnati Children’s Hospital Medical Center and University of Cincinnati College of Medicine, Cincinnati, OH, USA

    Michael V. Miles

Authors
  1. Peter H. Tang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael V. Miles
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael V. Miles .

Editor information

Editors and Affiliations

  1. , Molecular and Human Genetics, Baylor College of Medicine, 1 Baylor Plaza, Houston, 77030, Texas, USA

    Lee-Jun C. Wong, Ph.D.

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Tang, P.H., Miles, M.V. (2012). Measurement of Oxidized and Reduced Coenzyme Q in Biological Fluids, Cells, and Tissues: An HPLC-EC Method. In: Wong, Ph.D., LJ. (eds) Mitochondrial Disorders. Methods in Molecular Biology, vol 837. Humana Press. https://doi.org/10.1007/978-1-61779-504-6_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-504-6_10

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-503-9

  • Online ISBN: 978-1-61779-504-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation