Chromatographia

, Volume 66, Issue 7–8, pp 539–544 | Cite as

Analysis of Follicle-Stimulating Hormone by CE with Chemiluminescence Detection Based on Competitive Immunoassay

Original
First Online:
Received:
Revised:
Accepted:

Abstract

A competitive immunoassay and capillary electrophoresis with enhanced chemiluminescence detection have been used for determination of follicle-stimulating hormone (FSH) in human serum. The method is based on the competitive immunochemical reaction of FSH and horseradish peroxidase (HRP)-labeled FSH with anti-FSH, CE separation of antibody-bound and free HRP-labeled FSH, then chemiluminescence detection. The detection limit depends on the stability of the immune complex, which depends on analysis time and detector design, and on the chemiluminescence enhancer used. A unique chemiluminescence detector without dead volume or diluent effects was therefore used, and sodium tetraphenylboron was selected as the optimum enhancer. As a result sensitivity was substantially improved. Free HRP-labeled FSH and the immune complex could be separated within 15 min in alkaline borate buffer by use of a potential of 15 kV. Under the optimum conditions a calibration plot for FSH was established in the concentration range 0–100 mIU mL−1; the detection limit was 0.06 mIU mL−1. The concentration sensitivity achieved was 30 times better than that of ELISA.

Keywords

Capillary electrophoresis Chemiluminescence Competitive immunoassay Follicle-stimulating hormone Human serum 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgements

This work was supported by the Natural Science Foundation of China (Grant No 30470886).

References

  1. 1.
    Dobson H (1978) J Reprod Fertil 52:45–49CrossRefGoogle Scholar
  2. 2.
    Bolt DJ, Rollins R (1983) J Anim Sci 56:146–154Google Scholar
  3. 3.
    McNeilly JR, Fordyce M, Land R, Martin GB, Springbett AJ, Webb R (1988) J Reprod Fertil 84:213–221CrossRefGoogle Scholar
  4. 4.
    Rosenbusch J, Dias JA, Hodges JK (1997) Am J Primatol 41:179–193CrossRefGoogle Scholar
  5. 5.
    Qiu Q, Kuo A, Todd H, Dias JA, Gould JE, Overstreet JW, Lasley BL (1998) Fertil Steril 69:278–285CrossRefGoogle Scholar
  6. 6.
    Pappa A, Seferiadis K, Marselos M, Tsolas O, Messirtis IE (1999) Theriogenology 51:911–926CrossRefGoogle Scholar
  7. 7.
    Todd HE, Shideler SE, Laughlin LS, Overstreet JW, Pohl CR, Byrd W, Lasley BL (1999) Am J Primatol 48:135–151CrossRefGoogle Scholar
  8. 8.
    Prakash BS, Wallenhorst S, Metten M, Holtz W, Wuttke W (1999) Anim Reprod Sci 55:183–192CrossRefGoogle Scholar
  9. 9.
    Dighe AS, Moy JM, Hayes FJ, Sluss PM (2005) Clin Biochem 38:175–179CrossRefGoogle Scholar
  10. 10.
    Brindle E, Miller R, Shofer JB, Klein NA, Soules MR, O’Connor KA (2006) Clin Biochem 39:1071–1079CrossRefGoogle Scholar
  11. 11.
    Abad A, Moreno MJ, Pelegrí R, Martínez MI, Sáez A (1999) J Chromatogr A 833:3–12CrossRefGoogle Scholar
  12. 12.
    Vale P, Sampayo MA de M (1999) Toxicon 37:1565–1577CrossRefGoogle Scholar
  13. 13.
    Comper WD, Jerums G, Osicka TM (2004) Clin Biochem 37:105–111CrossRefGoogle Scholar
  14. 14.
    Goryacheva IYu De Saeger S, Lobeau M, Eremin SA, Barna-Vetró I, Van Peteghem C (2006) Anal Chim Acta 577:38–45CrossRefGoogle Scholar
  15. 15.
    Loureiro RF, de Oliveira JE, Torjesen PA, Bartolini P, Ribela MTCP (2006) J Chromatogr A 1136:10–18CrossRefGoogle Scholar
  16. 16.
    Bao JJ (1997) J Chromatogr B 699:463–480CrossRefGoogle Scholar
  17. 17.
    Schmalzing D, Buonocore S, Piggee C (2000) Electrophoresis 21:3919–3910CrossRefGoogle Scholar
  18. 18.
    Yeung WSB, Luo GA, Wang QG, Ou JP (2003) J Chromatogr B 797:217–228CrossRefGoogle Scholar
  19. 19.
    German I, Kennedy RT (2000) J Chromatogr B 742:353–362CrossRefGoogle Scholar
  20. 20.
    Su P, Zhang XX, Chang WB (2000) Electrophoresis 24:3197–3201CrossRefGoogle Scholar
  21. 21.
    Han KY, Ban E, Yoo YS (2003) J Chromatogr A 1013:215–220CrossRefGoogle Scholar
  22. 22.
    Yang WC, Yeung ES, Schmerr MJ (2005) Electrophoresis 26:1751–1759CrossRefGoogle Scholar
  23. 23.
    Xiao H, Li X, Zou H, Yang L, Yang Y, Wang Y, Wang H, Le XC (2006) Anal Chim Acta 556:340–346CrossRefGoogle Scholar
  24. 24.
    Wang JN, Ren JC (2005) Electrophoresis 26:2402–2406CrossRefGoogle Scholar
  25. 25.
    Wang JH, Huang WH, Liu YM, Cheng JK, Yang J (2004) Anal Chem 76:5393–5398CrossRefGoogle Scholar
  26. 26.
    Ji XH, He ZD, Ai XP, Yang HX, Xu CL (2006) Talanta 70:353–367CrossRefGoogle Scholar
  27. 27.
    Lin JH, Yan F, Ju HX (2004) Clin Chim Acta 341:109–115CrossRefGoogle Scholar
  28. 28.
    Thormann W, Molteni S, Caslavska J, Schmutz A (1994) Electrophoresis 15:3–12CrossRefGoogle Scholar
  29. 29.
    Zhou SY, Zhang ZJ (1996) Chin J Anal Chem 24:877–881CrossRefGoogle Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlag/GWV Fachverlage GmbH 2007

Authors and Affiliations

  1. 1.Institute Analytical ScienceSouthwest UniversityChongqingPeople’s Republic of China
  2. 2.Qiongzhou UniversityWuzhishanPeople’s Republic of China

Personalised recommendations