, Volume 31, Issue 6, pp 1043–1049 | Cite as

Citrate analysis using capillary electrophoresis and complexation with Eu3+-tetracycline

  • Douglas B. CraigEmail author
  • Jennifer R. Lischynski
  • Isabela C. C. Cardoso


A sensitive assay for citrate was developed. Citrate was incubated with 50 μM Eu3+-tetracycline and the complex separated using capillary electrophoresis utilizing post-column laser-induced luminescence detection in a sheath flow cuvette. Signal was linear with citrate concentration from 10 μM to 200 nM. Injection volumes were 320 pL. For the 200 nM sample, this corresponds to the injection of 64 amoles of citrate. Separation time was < 90 s with a total run time of 5 min. As an application the method was used to analyze citrate in agricultural and medicinal products. The method was also used to develop an assay for the enzyme citrate synthase.


Capillary electrophoresis Citrate Citrate synthase Eu3+-tetracycline Luminescence 



This study was supported by a Grant from the Natural Sciences and Engineering Research Council.


  1. Alam A, Kamruzzaman M, Lee SH, Kim YH, Jo HJ, Kim SH, Park S-R (2012) Sensitive determination of adenosine disodium triphosphate in soil, milk, and pharmaceutical formulation by enoxacin-europium (III) fluorescence complex in solution. J Lumin 132:789–794CrossRefGoogle Scholar
  2. Apleblat A (2014) Citric Acid, Springer, ISBN 978-3-319-11232-9Google Scholar
  3. Araujo CL, Melo EI, Coelho NMM (2011) Potentiometric detection of citrate in beverages using a graphite carbon electrode. Talanta 84:1169–1173CrossRefGoogle Scholar
  4. Arnaud N, Georges J (1999) Improved detection of salicylic acids using terbium-sensitized luminescence in aqueous micellar solutions of cetrytrimethylammonium chloride. Analyst 124:1075–1078CrossRefGoogle Scholar
  5. Cano-Raya C, Ramos MDF, Vallvey LFC, Wolfbeis OS, Schaferling M (2005) Fluorescencequenching of the europium tetracycline hydrogen peroxide complex by copper (II) and other metal ions. Appl Spectrosc 59:1209–1216CrossRefGoogle Scholar
  6. Chalgeri A, Tan HSI (1993) Assay of citrate in pharmaceutical matrices by indirect photometric liquid chromatography. J Pharm Biomed Anal 11:353–359CrossRefGoogle Scholar
  7. Chen G (2008) Portable tetracycline analyzer based on LED-excitation europium-sensitized luminescence. Sens Actuators, B 130:750–757CrossRefGoogle Scholar
  8. Chen DY, Dovichi NJ (1994) Yoctomole detection limit by laser-induced fluorescence in capillary electrophoresis. J Chromatogr B 657:265–269CrossRefGoogle Scholar
  9. Chen DY, Dovichi NJ (1996) Single-molecule detection in capillary electrophoresis: molecular shot noise as a fundamental limit to chemical analysis. Anal Chem 68:690–696CrossRefGoogle Scholar
  10. Chen DY, Adelhelm K, Cheng XL, Dovichi NJ (1994) A simple laser-induced fluorescence detector for sulforhodamine 101 in a capillary electrophoresis system: detection limits of 10 yoctomoles or six molecules. Analyst 119:349–352CrossRefGoogle Scholar
  11. Chen GY, Schneider MJ, Darwish AM, Lehotay SJ, Freeman DW (2004) Europium sensitized luminescence determination of oxytetracycline in catfish muscle. Talanta 64:252–257CrossRefGoogle Scholar
  12. Craig DB, Hiebert Z (2017) Analysis of complexes of metabolites with europium tetracycline using capillary electrophoresis coupled with laser-induced luminescence detection. Biometals 30:449–458CrossRefGoogle Scholar
  13. DaSilva FR, Courrol LC, Tarelho LVG, Gomes L, Vieira ND (2005) Enhancement of europium luminescence in tetracycline-europium complexes in the presence of urea hydrogen peroxide. J Fluoresc 15:667–671CrossRefGoogle Scholar
  14. DeBorba BM, Rohrer JS, Bhattacharyya L (2004) Development and validation of an assay for citric acid/citrate and phosphate in pharmaceutical dosage forms using ion chromatography with suppressed conductivity detection. J Pharm Biomed Anal 36:517–524CrossRefGoogle Scholar
  15. Duerkop A, Turel M, Lobnik A, Wolfbeis OS (2006) Microtiter plate assay for phosphate using a europium-tetracycline complex as a sensitive luminescent probe. Anal Chim Acta 555:292–298CrossRefGoogle Scholar
  16. Durkop A, Wolfbeis OS (2005) Nonenzymatic direct assay of hydrogen peroxide at neutral pH using the Eu3Tc fluorescent probe. J. Fluoresc 15:755–761CrossRefGoogle Scholar
  17. Holden AJ, Littlejohn D, Fell GS (1996) Determination of citrate in plasma protein solutions by UV-visible spectrophotometry and ion chromatography. J Pharm Biomed Anal 14:713–719CrossRefGoogle Scholar
  18. Hotha KK, Patel T, Roychowdhury S, Subramanian V (2014) Development of better-quality assay method for the citric acid and sodium citrate in ophthalmic/oral solutions and their application to deformulation studies. Am J Anal Chem 5:1249–1260CrossRefGoogle Scholar
  19. Jaworska M, Cygan P, Wilk M, Anuszewska E (2009) Capillary electrophoresis with indirect UV detection for the determination of stabilizers and citrates present in human albumin solutions. J Pharm Biomed Anal 50:90–95CrossRefGoogle Scholar
  20. Jiang CQ, Hou FJ (2004) Fluorescence study of the interaction between a modified β-cyclodextrin and metacycline-Eu3+ and its analytical application. Anal Lett 37:1885–1895CrossRefGoogle Scholar
  21. Jiang CQ, Luo L (2004) Spectrofluorometric determination of human serum albumin using a tetracycline-europium complex. Anal Lett 37:1129–1137CrossRefGoogle Scholar
  22. Kaleemullah T, Ahmed M, Sharma HK, Rao MN (2011) Sensitive ion chromatographic determination of citrate and formate in pharmaceuticals. Rasayan J Chem 4:844–852Google Scholar
  23. Kautenburger R, Beck HP (2007) Complexation studies with lanthanides and humic acid analyzed by ultrafiltration and capillary-electrophoresis-inductively coupled plasma mass spectrometry. J Chromatogr 1159:75–80CrossRefGoogle Scholar
  24. Kozhevnikov VN, Mandl C, Miltschitzky S, Durkop A, Wolfbeis OS, Koenig B (2005) Strong emission increase of a dicarboxyterpyridene europium(III) complex in the presence of citrate and hydrogen peroxide. Inorganica Chim Acta 358:2445–2448CrossRefGoogle Scholar
  25. Latva M, Ala-Kleme T, Bjennes H, Kankare J, Haapakka K (1995) Time-resolved luminescence detection of europium(III) chelates in capillary electrophoresis. Analyst 120:367–372CrossRefGoogle Scholar
  26. Lin Z, Wu M, Schaeferling M, Wolfbeis OS (2004) Fluorescent imaging of citrate and other intermediates in the citric acid cycle. Angew Chemie 43:1735–1738CrossRefGoogle Scholar
  27. Lin Z, Wu M, Wolfbeis OS (2005) Time-resolved fluorescent chirality sensing and imaging of malate in aqueous solution. Chirality 17:464–469CrossRefGoogle Scholar
  28. Liu Z-H, Devaraj S, Yang C-R, Yen Y-P (2012) A new selective chromogenic and fluorogenic sensor for citrate ion. Sens Actuators 174:555–562CrossRefGoogle Scholar
  29. Luo LY, Luo L, Luo YH, Wang CY, Jiang C, Chen Q, Luo M, Long F, Wang S, Tan MG, Chen D (2013) Determination of potassium citrate and sodium citrate chewable tablets content and related substances by RP-HPLC. Lat Am J Pharm 32:37–42Google Scholar
  30. McCleskey SC, Metzger A, Simmons CS, Anslyn EV (2002) Competitive indicator methods for the analysis of citrate using colorimetric assays. Tetrahedron 58:621–626CrossRefGoogle Scholar
  31. Peng Q, Hou F, Ge X, Jiang C, Gong S (2005) Fluorimetric study of the interaction between nicotinamide adenine dinucleotide phosphate and tetracycline-europium complex and its application. Anal Chim Acta 549:26–31CrossRefGoogle Scholar
  32. Peng Q, Ge X, Jiang C (2007) A new spectrofluorometric probe for the determination of trace amounts of CoA in injection, human serum and pig livers. Anal Sci 23:557–561CrossRefGoogle Scholar
  33. Penniston KL, Nakada SY, Holmes RP, Assimos DG (2008) Quantitative assessment of citric acid in lemon juice, lime juice, and commercially-available fruit juice products. J Endourol 22(3):567–570CrossRefGoogle Scholar
  34. Rodriguez RC, Aguilar-Caballos MP, Gomez-Hens A (2003) Simultaneous determination of ciprofloxin and tetracycline in biological fluids based on dual-lanthanide sensitized luminescence using dry reagent chemical technology. Anal Chim Acta 494:55–62CrossRefGoogle Scholar
  35. Schaferling M, Wolfbeis OS (2007) Europium tetracycline as a luminescent probe for nucleoside phosphates and its application to the determination of kinase activity. Chemistry 15:4342–4349CrossRefGoogle Scholar
  36. Shapiro F, Silanikove N (2011) Rapid and accurate determination of malate, citrate, pyruvate and oxaloacetate by enzymatic reactions coupled to formation of a fluorochromophore: application in colorful juices and fermentable food (yogourt, wine) analysis. Food Chem 129:608–613CrossRefGoogle Scholar
  37. Shriver D, Weller M, Overton T, Rourke J, Armstrong F (2014) Inorganic chemistry, 6th edn. WH Freeman and Company, New YorkGoogle Scholar
  38. Shtykov SN, Smirnova TD, Bylinkin YG, Zhemerichkin DA (2005) Fluorimetric determination of tetracyclines with europium chelates of 1,10-phenanthroline in micellar soultions of anionic surfactants. J Anal Chem 60:24–28CrossRefGoogle Scholar
  39. Stathakis C, Cassidy RM (1996) Capillary electrophoretic separations of metal ions in the presence of polyethylene glycols. Analyst 121:839–843CrossRefGoogle Scholar
  40. Sun Y, Xiao F, Zhong C, Xue P, Fu A (2014) Colorimetric sensing ensemble for citrate in water. Sens Actuators, B 194:269–275CrossRefGoogle Scholar
  41. Verma SP, Garcia R, Santoyo E, Aparico A (2000) Improved capillary electrophoresis method for measuring rare-earth elements in synthetic geochemical standards. J Chromatogr A 884:317–328CrossRefGoogle Scholar
  42. Voet D, Voet JG, Pratt CW (2016) Fundamentals of biochemistry: life at the molecular level, 5th edn. Wiley, HobokenGoogle Scholar
  43. Wang T, Jiang CQ (2006) Spectrofluorimetric determination of lecithin using a tetracycline-europium probe. Anal Chim Acta 561:204–209CrossRefGoogle Scholar
  44. Yamaguchi Y, Hashino K, Ito M, Ikawa K, Nishioka T, Matsumoto K (2009) Sodium dodecyl sulfate polyacrylamide slab gel electrophoresis and hydroxyethyl cellulose gel capillary electrophoresis of luminescent lanthanide chelate-labeled proteins with time-resolved detection. Anal Sci 25:327–332CrossRefGoogle Scholar
  45. Yegorova A, Vityukova E, Beltyukova S, Durkop A (2006) Determination of citrate in tablets and of oxytetracycline in serum using europium (III) luminescence. Microchem J 83:1–6CrossRefGoogle Scholar
  46. Zhu XJ, Wang XL, Jiang CQ (2005) Spectrofluorometric determination of heparin using a tetracycline-europium probe. Anal Biochem 341:299–307CrossRefGoogle Scholar
  47. Zhu Z, Zhou J, Li Z, Yang C (2015) Dinuclear zinc complex for fluorescent indicator-displacement assay of citrate. Sens Actuators, B 208:151–158CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Douglas B. Craig
    • 1
    Email author
  • Jennifer R. Lischynski
    • 1
  • Isabela C. C. Cardoso
    • 1
  1. 1.Department of ChemistryUniversity of WinnipegWinnipegCanada

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