Microchimica Acta

, Volume 166, Issue 3–4, pp 243–249

Sensitive and rapid determination of nitric oxide in human serum using microchip capillary electrophoresis with laser-induced fluorescence detection

Original Paper

Abstract

Microfluidic chip capillary electrophoresis with laser-induced fluorescence detection is employed for direct determination of trace nitric oxide in human blood using diaminorhodamines as the fluorescence probe. Factors influencing the separation and detection processes were systematically studied. Complete and fast separation of the highly fluorescent triazole formed was achieved within 45 s, and the relative standard deviations values of migration time and peak area were less than 3%. The detection limit of NO was 3.0 nmol.L-1 (at a signal-to-noise ratio of 3) and the liner range was from 1.0 × 10-8 mol.L-1 to 3.0 × 10-6 mol.L-1. The method has been applied to the determination of NO in serum of healthy persons and patients suffering from diseases, with recoveries varying from 92.65 to 98.43%.

Keywords

Nitric oxide DAR-4M Microchip capillary electrophoresis Laser-induced fluorescence detection 

References

  1. 1.
    Murad F (2006) Nitric oxide and cyclic GMP in cell signaling and drug development. N Engl J Med 355:2003CrossRefGoogle Scholar
  2. 2.
    Garthwaite J, Boulton CL (1995) Nitric oxide signaling in the central nervous system. Ann Rev Physiol 57:683CrossRefGoogle Scholar
  3. 3.
    Jaeschke H, Schini VB, Farhood A (1992) Role of nitric oxide in the oxidant stress during ischemia/reperfusion injury of the liver. Life Sci 50:1797CrossRefGoogle Scholar
  4. 4.
    Lauer T, HeiB C, Strauer BE, Feelisch M, Kelm M (2002) Evidence for in vivo transport of bioactive nitric oxide in human plasma. J Clin Invest 109:1241Google Scholar
  5. 5.
    Berkels R, Purol-Schnabel S, Roesen R (2001) A new method to measure nitrate/nitrite with a NO-sensitive electrode. J Appl Physiol 90:317Google Scholar
  6. 6.
    Sergio AN, Luis AJR, Martha OSM, Joel RE, Eva EA, Patricia CM (2004) Indirect determination of nitric oxide production by reduction of nitrate with a freeze-thawing-resistant nitrate reductase from Escherichia coli MC1061. Anal Biochem 328:14CrossRefGoogle Scholar
  7. 7.
    Zhou XJ, Arnold MA (1996) Response characteristics and mathematical modeling for a nitric oxide fiber-optic chemical sensor. Anal Chem 68:1748CrossRefGoogle Scholar
  8. 8.
    Bedwell DW, Rivera VR, Merrill GA, Pusateri AE (2000) Elimination of matrix-based interferences to a fluorescent nitrite/nitrate assay by a simple filtration procedure. Anal Biochem 284:1CrossRefGoogle Scholar
  9. 9.
    Yao DD, Vlessiolis AG, Evimirids NP (2004) Determination of Nitric Oxide in Biological Samples. Microchim Acta 147:1CrossRefGoogle Scholar
  10. 10.
    Chen XX, Wang Y, Sheng SH (2008) A novel amperometric sensor for the determination of nitric oxide, and its application in rat liver cells. Microchim Acta 161:255CrossRefGoogle Scholar
  11. 11.
    Isik S, Oni J, Rjabova V, Neugebauer S, Schuhmann W (2004) Entrapment of metalloporphyrins within an electrodeposition paint layer as a basis for developing a nitric oxide sensor. Microchim Acta 148:59CrossRefGoogle Scholar
  12. 12.
    He Q, Zheng DY, Hu SS (2009) Development and application of a nano-alumina based nitric oxide sensor. Microchim Acta 164:459CrossRefGoogle Scholar
  13. 13.
    Huang KJ, Wang H, Zhang QY, Ma M, Hu JF, Zhang HS (2006) Direct detection of nitric oxide in human blood serum by use of 1, 3, 5, 7-tetramethyl-8-(3, 4-diaminophenyl)difluoroboradiaza- s-indacene with HPLC. Anal Bioanal Chem 384:1284CrossRefGoogle Scholar
  14. 14.
    Huang KJ, Zhang M, Zhang HS, Wang H (2007) Sensitive determination of ultra-trace nitric oxide in blood using derivatization-polymer monolith microextraction coupled with reversed-phase high-performance liquid chromatography. Anal Chim Acta 591:116CrossRefGoogle Scholar
  15. 15.
    Dittrich PS, Tachikawa K, Manz A (2006) Micro total analysis systems. Latest advancements and trends. Anal Chem 78:3887CrossRefGoogle Scholar
  16. 16.
    Miyado T, Takeda Y, Nagai H, Saito K, Fukushi K, Yoshida Y, Wakida S, Niki E (2003) High-throughput nitric oxide assay in biological fluids using microchip capillary electrophoresis. J Chromatgr A 1109:174CrossRefGoogle Scholar
  17. 17.
    Kojima H, Nakatsubo N, Kikuchi K, Kawahara S, Kirino Y, Nagoshi H, Hirata Y, Nagano T (1998) Detection and imaging of nitric oxide with novel fluorescent indicators: diaminofluoresceins. Anal Chem 70:2446CrossRefGoogle Scholar
  18. 18.
    Kojima H, Hirotani M, Nakatsubo N, Kikuchi K, Kirino Y, Urano Y, Higuchi T, Nagoshi H, Hirata Y, Nagano T (2001) Bioimaging of nitric oxide with fluorescent indicators based on the rhodamine chromophore. Anal Chem 73:1967CrossRefGoogle Scholar
  19. 19.
    Dacres H, Nareyanswanry R (2005) Evaluation of 1, 2-diaminoanthraquinone (DAA) as a potential reagent system for detection of NO. Microchim Acta 152:35CrossRefGoogle Scholar
  20. 20.
    Gabe Y, Urano Y, Kikuchi K, Kojima H, Nagano T (2004) Highly sensitive fluorescence probes for nitric oxide based on boron dipyrromethene chromophore-rational design of potentially useful bioimaging fluorescence probe. J Am Chem Soc 126:3357CrossRefGoogle Scholar
  21. 21.
    Lu WJ, Zhang YL, Yin M (2007) Fabrication of the master for soft lithography by electron beam lithography. J Optoelectronics·Laser 18:19Google Scholar
  22. 22.
    McDonald J, Duffy D, Anderson J, Chiu D, Wu H, Schueller O, Whitesides G (2000) Fabrication of microfluidic systems in poly(dimethylsiloxane). Electrophoresis 21:27CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.College of Chemistry, Chemical Engineering and Materials ScienceShandong Normal UniversityJinanChina
  2. 2.Institute of Electron Beam, School of Control Science and EngineeringShandong UniversityJinanChina

Personalised recommendations