Analytical and Bioanalytical Chemistry

, Volume 400, Issue 2, pp 395–401 | Cite as

SPE/RP-HPLC using C1 columns: an environmentally friendly alternative to conventional reverse-phase separations for quantitation of beta-carboline alkaloids in human serum samples

  • Víctor González-Ruiz
  • Ana I. Olives
  • M. Antonia MartínEmail author
Original Paper


The analysis of beta-carboline alkaloids presents a renewed interest due to their biological relevance and their increasing popularity as recreational drugs. In the present work, a novel chromatographic reverse-phase high-performance liquid chromatography (RP-HPLC) method with fluorimetric detection has been applied to the determination of beta-carbolines spiked in human serum samples. The chromatographic procedure involves the use of less retentive, unusual C1 columns combined with hydro-alcoholic mobile phases and the use of beta-cyclodextrin or (2-hydroxypropyl)-beta-cyclodextrin as mobile-phase additives. The effective combination of C1 columns and the modified mobile phases with cyclodextrins leads to a considerable reduction in the organic proportion in the mobile phase (up to 50%) with good resolution and efficiency. Besides, the presence of cyclodextrins allows the use of ethanol, a green solvent, as the organic component in the mobile phase. Traditional RP-HPLC thus becomes an attractive eco-separation technique using conventional stationary phases under simple and user-friendly experimental conditions. Solid-phase extraction was employed as sample clean-up protocol with attractive features, i.e., a low consumption of organic solvents, time and step economy and diminished need for sample handling. The analytical procedure was completely validated showing satisfactory figures of merit. Limits of detection of 10−9–10−10 M can be achieved. The recoveries obtained for the total methodology (sample pre-treatment and chromatographic determination in the case of the mobile phases containing cyclodextrins) were very satisfactory (95–107%) as well as the intraday (2–3%) and interday precision values (3–7%). The use of 3-hydroxymethyl-beta-carboline as an internal standard allows the comparison of the goodness of response of the analytical methodology in the presence or absence of cyclodextrins.


Cyclodextrins Eco-friendly mobile phases β-carboline derivatives HPLC 











High-performance liquid chromatography


Internal standard


Mobile-phase additives


Quality control samples


Reverse phase


Solid-phase extraction



Financial support from Ministerio de Ciencia e Innovación (SPAIN) through grant CTQ2009-11312 as well as from Grupos de investigación UCM grant 920234 is gratefully acknowledged. The authors are grateful to MEC for a FPU research fellowship for V. González-Ruiz.

Supplementary material

216_2010_4562_MOESM1_ESM.pdf (431 kb)
ESM 1 (PDF 430 kb)


  1. 1.
    Casal S, Mendes E, Fernandes JO, Oliveira MBPP, Ferreira MA (2004) J Chromatogr A 1040:105–114CrossRefGoogle Scholar
  2. 2.
    Stöckigt J, Sheludko Y, Unger M, Gerasimenko I, Warzecha H, Stöckigt D (2002) J Chromatogr B 802:115–126Google Scholar
  3. 3.
    Hemmateenejad B, Abbaspour A, Maghami H, Miri R, Panjehshahin MR (2006) Anal Chim Acta 575:290–299CrossRefGoogle Scholar
  4. 4.
    Zheng W, Wang S, Barnes LF, Guan Y, Louis ED (2000) Anal Biochem 279:125–129CrossRefGoogle Scholar
  5. 5.
    Herraiz T, Chaparro C (2006) J Chromatogr A 1120:237–243CrossRefGoogle Scholar
  6. 6.
    Aguí L, Peña-Farfal C, Yáñez-Sedeño P, Pingarrón JM (2007) Anal Chim Acta 585:323–330CrossRefGoogle Scholar
  7. 7.
    Smith CJ, Qian X, Zha Q, Moldoveanu SC (2004) J Chromatogr A 1046:211–216Google Scholar
  8. 8.
    Herraiz T, Chaparro C (2005) Biochem Biophys Res Commun 326:378–386CrossRefGoogle Scholar
  9. 9.
    Pfau W, Skog K (2004) J Chromatogr A 1046:211–216Google Scholar
  10. 10.
    Frison G, Favretto D, Zancanaro F, Fazzin G, Ferrara SD (2008) Foresic Sci Int 179:e37–e43CrossRefGoogle Scholar
  11. 11.
    Herraiz T, González D, Ancín-Azpilicueta C, Arán VJ, Guillén H (2010) Food Chem Toxicol 48:839–845CrossRefGoogle Scholar
  12. 12.
    Callaway JC (2005) J Psichoact Drugs 37:151–155Google Scholar
  13. 13.
    Inman WD, Bray WM, Gassner NC, Lokey RS, Tenney K, Shen YY, Tendyke K, Suh T, Crews P (2010) J Nat Prod 73:255–257CrossRefGoogle Scholar
  14. 14.
    Charan RD, McKee TC, Gustafson KR, Pannell LK, Boyd MR (2002) Tetrahedron Lett 43:5201–5204CrossRefGoogle Scholar
  15. 15.
    Chouvenc T, Su N, Elliot M (2008) J Invertebr Pathol 99:345–347CrossRefGoogle Scholar
  16. 16.
    Tsuchiya H (2004) J Chromatogr A 1031:325–330CrossRefGoogle Scholar
  17. 17.
    Mckenna DJ (2004) Pharmacol Ther 102:11–129CrossRefGoogle Scholar
  18. 18.
    Pieroni A, Muenzc H, Akbulut M, Baser KH, Durmuskahya C (2005) J Ethnopharmacol 102:69–88CrossRefGoogle Scholar
  19. 19.
    Boyer EW, Shannon M, Hibberd PL (2001) N Engl J Med 345:469–471CrossRefGoogle Scholar
  20. 20.
    Welch CJ, Sajonz P, Biba M, Gouker J, Fairchild J (2006) J Liq Chromatogr 29:2185–2200CrossRefGoogle Scholar
  21. 21.
    Sajonz P, Schafer W, Gong X, Schultz S, Rosner T, Welch CJ (2007) J Chromatogr A 1147:149–154CrossRefGoogle Scholar
  22. 22.
    Jandera P, Bocian S, Molíková M, Buszewski B (2009) J Chromatogr A 1216:237–248CrossRefGoogle Scholar
  23. 23.
    Zhu L, Ding L, Zhang Q, Wang L, Tang F, Liu Q, Yao S (2009) Green Chem 11:132–137CrossRefGoogle Scholar
  24. 24.
    El-Gindy A, Emara S, Mesbah MK, Hadad GM (2006) J Assoc Off Anal Chem 89:65–70Google Scholar
  25. 25.
    López-Nicolás JM, Núñez-Delicado E, Pérez-López AJ, Carbonell-Barrachina A, Cuadra-Crespo P (2006) J Chromatogr A 1135:158–165CrossRefGoogle Scholar
  26. 26.
    López-Nicolás JM, García-Carmona F (2008) Food Chem 109:868–875CrossRefGoogle Scholar
  27. 27.
    Sezjtli J (1998) Chem Rev 98:1743–1753CrossRefGoogle Scholar
  28. 28.
    Cserháti T, Forgacs E (2003) Cyclodextrins in chromatography. Royal Society of Chemistry, CambridgeGoogle Scholar
  29. 29.
    Capelo C, Fisher U, Hungerbühler K (2007) Gren Chem 9:927–934CrossRefGoogle Scholar
  30. 30.
    Welch CJ, Brkovic T, Schafer W, Gong X (2007) Green Chem 11:1232–1238CrossRefGoogle Scholar
  31. 31.
    Moffat AC, Jackson JV, Moss SM, Widdop B (1986) Clarke’s isolation and identification of drugs. Pharmaceutical Press, LondonGoogle Scholar
  32. 32.
    Adachi J, Mizoi Y, Naito T, Yamamoto K, Fujiwara S, Ninomiya I (1991) J Chromatogr 531:331–339CrossRefGoogle Scholar
  33. 33.
    Food and Drug Administration (2001) Guidance for Industry—Bioanalytical Method Validation. US Department of Health and Human Services, RockvilleGoogle Scholar
  34. 34.
    International Conference on Harmonisation (1996) ICH Topic Q 2 B. Validation of analytical procedures methodologyGoogle Scholar
  35. 35.
    Thompson M, Ellison SLR, Wood R (2002) Pure Appl Chem 74:835–855CrossRefGoogle Scholar
  36. 36.
    Kerton FM (2009) Alternative solvents for green chemistry. Royal Society of Chemistry, CambridgeGoogle Scholar
  37. 37.
    León AG, Olives AI, Martín MA, del Castillo B (2007) J Incl Phenom Macrocycl Chem 57:577–583CrossRefGoogle Scholar
  38. 38.
    Tsuchiya H, Shimizu H, Iimuna M (1999) Chem Pharm Bull 47:440–443Google Scholar
  39. 39.
    Louis ED, Zheng W, Jurewicz EC, Watner D, Chen J, Factor-litvak P, Parides M (2002) Neurology 59:1940–1944Google Scholar
  40. 40.
    Yritia M, Riba J, Ortuño J, Ramírez A, Castillo A, Alfaro Y, de la Torre R, Barbanoj MJ (2002) J Chromatogr B 779:271–281CrossRefGoogle Scholar
  41. 41.
    Back YM, Lee JH, Shin HS, Lee KG (2009) Food Addit Contam 26:298–305CrossRefGoogle Scholar
  42. 42.
    Kartal M, Altun M, Kurucu S (2003) J Pharm Biomed Anal 31:263–269CrossRefGoogle Scholar
  43. 43.
    Herraiz T, Guillén H, Arán VJ (2008) Chem Res Toxicol 21:2172–2180CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Víctor González-Ruiz
    • 1
  • Ana I. Olives
    • 1
  • M. Antonia Martín
    • 1
    Email author
  1. 1.Sección Departamental de Química Analítica, Facultad de FarmaciaUniversidad Complutense de MadridMadridSpain

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