Skip to main content
SpringerLink
Log in

Determination of Cytochrome c in Human Serum and Pharmaceutical Injections Using Flow Injection Chemiluminescence

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

It was found that the complex of cytochrome c (Cyt c) and hydrogen peroxide could significantly catalyze the chemiluminescence (CL) reaction from luminol–hydrogen peroxide, and a sensitive, rapid, and simple CL procedure was proposed for the determination of Cyt c in a flow injection system for the first time. The increment of CL intensity was linear over the concentration of Cyt c ranging from 5 to 700 ng ml−1, with a detection limit of 2 ng ml−1 (3σ). At a flow rate of 2.0 ml min−1, a complete analytical process could be performed in 30 s with a relative standard deviation of less than 4.0%. The proposed method was applied successfully for the assay of Cyt c in pharmaceutical injections and human serum, and the recoveries were from 98.0% to 108.8% and 92.5% to 109.0%. The possible mechanism of Cyt c enhanced CL reaction was also discussed.

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. Raphael, A. L., & Gray, H. B. (1991). Semisynthesis of axial-ligand (position 80) mutants of Cytochrome c. Journal of the American Chemical Society, 113, 1038–1040. doi:10.1021/ja00003a045.

    Article  CAS  Google Scholar 

  2. Jiang, X., & Wang, X. (2004). Cytochrome c-mediated apoptosis. Annual Review of Biochemistry, 73, 87–106. doi:10.1146/annurev.biochem.73.011303.073706.

    Article  CAS  Google Scholar 

  3. Green, D. R., & Kroemer, G. (2004). The pathophysiology of mitochondrial cell death. Science, 305, 626–629. doi:10.1126/science.1099320.

    Article  CAS  Google Scholar 

  4. Xua, J. M., Li, W., Yin, Q. F., & Zhu, Y. L. (2007). Direct electrochemistry of cytochrome c on natural nano-attapulgite clay modified electrode and its electrocatalytic reduction for H2O2. Electrochimica Acta, 52, 3601–3606. doi:10.1016/j.electacta.2006.10.020.

    Article  Google Scholar 

  5. Ehrlichova, M., Koc, M., Truksa, J., Naldova, Z., Vaclavikova, R., & Kovarr, J. (2005). Cell death induced by taxanes in breast cancer cells: Cytochrome c is released in resistant but not in sensitive cells. Anticancer Research, 25, 4215–4224.

    CAS  Google Scholar 

  6. Lin, H. I., Lee, Y. J., Chen, B. F., Tsai, M. C., Lu, J. L., Chou, C. J., et al. (2005). Involvement of Bcl-2 family, cytochrome c and caspase 3 in induction of apoptosis by beauvericin in human non-small cell lung cancer cells. Cancer Letters, 230, 248–259. doi:10.1016/j.canlet.2004.12.044.

    Article  CAS  Google Scholar 

  7. Petrosillo, G., Ruggiero, F. M., Pistolese, M., & Paradies, G. (2004). Attenuation of fatty acid-induced apoptosis by low-dose alcohol in neonatal rat cardiomyocytes. The Journal of Biological Chemistry, 279, 53103–53108. doi:10.1074/jbc.M407500200.

    Article  CAS  Google Scholar 

  8. Liu, Y. C., Sowdal, L. H., & Robinson, N. C. (1995). Separation and quantitation of cytochrome c oxidase subunits by Mono-Q fast protein liquid chromatography and C18 reverse-phase high-performance liquid chromatography. Archives of Biochemistry and Biophysics, 324, 135–142. doi:10.1006/abbi.1995.9917.

    Article  CAS  Google Scholar 

  9. Picklo, M. J., Zhang, J., Nguyen, V. Q., Graham, D. G., & Montine, T. J. (1999). High-pressure liquid chromatography quantitation of cytochrome c using 393 nm detection. Analytical Biochemistry, 276, 166–170. doi:10.1006/abio.1999.4349.

    Article  CAS  Google Scholar 

  10. Batthyány, C., Souza, J. M., Durán, R., Cassina, A., Cerveñansky, C., & Radi, R. (2005). Time course and site(s) of cytochrome c tyrosine nitration by peroxynitrite. Biochemistry, 44, 8038–8046. doi:10.1021/bi0474620.

    Article  Google Scholar 

  11. Meagher, R. J., Seong, J., Laibinis, P. E., & Barron, A. E. (2004). A very thin coating for capillary zone electrophoresis of proteins based on a tri(ethylene glycol)-terminated alkyltrichlorosilane. Electrophoresis, 25, 405–414. doi:10.1002/elps.200305714.

    Article  CAS  Google Scholar 

  12. Mohan, D., & Lee, C. S. (2002). On-line coupling of capillary isoelectric focusing with transient isotachophoresis-zone electrophoresis: A two-dimensional separation system for proteomics. Electrophoresis, 23, 3160–3167. doi:10.1002/1522-2683(200209) 23:18≤3160::AID-ELPS3160≥3.0.CO;2-L.

    Article  CAS  Google Scholar 

  13. Valle-Mondragón, L. D., Ramírez-Ortega, M., Zarco-Olvera, G., Sánchez-Mendoza, A., Pastelín- Hernández, G., & Tenorio-López, F. A. (2008). Capillary zone electrophoretic determination of cytochrome c in mitochondrial extracts and cytosolic fractions: Application to a digitalis intoxication study. Talanta, 74, 278–288.

    Google Scholar 

  14. Albrecht, H. O. (1928). Uber die Chemiluminescenz des Aminophthalsaurehydrazids. Zeitschrift für Physikalische Chemie, 136, 321–330.

    Google Scholar 

  15. Powe, A. M., Fletcher, K. A., St. Luce, N. N., Lowry, M., Neal, S., McCarroll, M. E., et al. (2004). Molecular fluorescence, phosphorescence, and chemiluminescence spectrometry. Analytical Chemistry, 76, 4614–4634. doi:10.1021/ac040095d.

    Article  CAS  Google Scholar 

  16. Fletcher, K. A., Fakayode, S. O., Lowry, M., Tucker, S. A., Neal, S. L., Kimaru, I. W., et al. (2006). Molecular fluorescence, phosphorescence, and chemiluminescence spectrometry. Analytical Chemistry, 78, 4047–4068. doi:10.1021/ac060683m.

    Article  CAS  Google Scholar 

  17. Song, Z. H., Liu, Y. H., & Xie, X. F. (2006). In vitro monitoring picogram roxithromycin in human urine using flow injection chemiluminescence procedure. Current Drug Metabolism, 7, 389–395. doi:10.2174/138920006776873481.

    Article  CAS  Google Scholar 

  18. Liu, Y. H., Song, Z. H., Dong, F. X., & Zhang, L. (2007). Flow injection chemiluminescence determination of Sudan I in hot chilli sauce. Journal of Agricultural and Food Chemistry, 55, 614–617. doi:10.1021/jf063332h.

    Article  CAS  Google Scholar 

  19. Xie, X. F., Shao, X. D., & Song, Z. H. (2006). Determination of risperidone at picogram level in human urine by luminol–H2O2 chemiluminescence. Chem. Pap., 60, 288–292. doi:10.2478/s11696-006-0050-8.

    Article  CAS  Google Scholar 

  20. Feissner, R., Xiang, Y. B., & Kranz, R. G. (2003). Chemiluminescent-based methods to detect subpicomole levels of c-type cytochromes. Analytical Biochemistry, 315, 90–94. doi:10.1016/S0003-2697(02) 00658-9.

    Article  CAS  Google Scholar 

  21. Zhu, Z. Y. (1992). Shiyong Yixue Jianyanxue (1st ed.), pp. 415–417. Beijing, China: People’s Military Medical Press.

    Google Scholar 

  22. Slawinski, J., Galezowski, W., & Elbanowski, M. (1981). Chemiluminescence in the reaction of cytochrome c with hydrogen peroxide. Biochimica et Biophysica Acta, 637, 130–137. doi:10.1016/0005-2728(81) 90218-8.

    Article  CAS  Google Scholar 

  23. Zhang, X. A., Wang, J. F., Wu, W. J., Qian, S. W., & Man, Y. H. (2007). Immobilization and electrochemistry of cytochrome c on amino-functionalized mesoporous silica thin films. Electrochemistry Communications, 9, 2098–2104. doi:10.1016/j.elecom.2007.05.025.

    Article  CAS  Google Scholar 

  24. Svistunenko, D. A., Wilson, M. T., & Cooper, C. E. (2004). Tryptophan or tyrosine? On the nature of the amino acid radical formed following hydrogen peroxide treatment of cytochrome c oxidase. Biochimica et Biophysica Acta, 1655, 372–380. doi:10.1016/j.bbabio.2003.06.006.

    Article  CAS  Google Scholar 

Download references

Acknowledgment

The authors gratefully acknowledge the financial support from Shaanxi Province Nature Science Foundation and Ministry of Education, China, grant no. 2006B05 and no. 07JK395.

Author information

Authors and Affiliations

  1. Key Laboratory of Synthetic and Natural Functional Molecule Chemistry of Ministry of Education, College of Chemistry & Material Science, Shaanxi Key Laboratory of Physico-Inorganic Chemistry, Northwest University, Xi’an, 710069, China

    Xin Li, Houyong Liu, Xili He & Zhenghua Song

Authors
  1. Xin Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Houyong Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xili He
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhenghua Song
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhenghua Song.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, X., Liu, H., He, X. et al. Determination of Cytochrome c in Human Serum and Pharmaceutical Injections Using Flow Injection Chemiluminescence. Appl Biochem Biotechnol 160, 1065–1073 (2010). https://doi.org/10.1007/s12010-009-8598-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-009-8598-8

Keywords

Search

Navigation