Analytical and Bioanalytical Chemistry

, Volume 400, Issue 9, pp 2903–2912 | Cite as

Sarcosine as a marker in prostate cancer progression: a rapid and simple method for its quantification in human urine by solid-phase microextraction–gas chromatography–triple quadrupole mass spectrometry

  • Brunella Cavaliere
  • Barbara Macchione
  • Marcello Monteleone
  • Attilio Naccarato
  • Giovanni Sindona
  • Antonio Tagarelli
Original Paper


Sarcosine is an amino acid derivative of N-methylglycine and is involved in the amino acid metabolism and methylation processes that are enriched during prostate cancer progression. It could also serve as a new target to be measured during therapeutic interventions and help in the identification of aggressive tumors for radical treatment. In this study, we present a new urine test that can help early diagnosis of prostate cancer. The method for the quantification of sarcosine in urine consists of a solid-phase microextraction (SPME) step followed by gas chromatography–triple quadrupole mass spectrometry analysis. We used a preliminary derivatization step with ethyl chloroformate/ethanol and the corresponding ester was then extracted by SPME in immersion mode. Several fibers were evaluated and the optimization of the parameters affecting the SPME process was carried out using an experimental design. The optimal values were 20 min extraction time, 10% NaCl, and 270°C using a divinylbenzene/Carboxen/polydimethylsiloxane fiber. The triple quadrupole analyzer acquired data in selected reaction monitoring mode, allowing us to obtain reconstructed chromatograms with well-defined chromatographic peaks. The accuracy and precision of this method were evaluated at concentrations of 70, 250, and 800 ng/ml and were found to be acceptable. Very satisfactory values (0.10 and 0.16 ng/ml, respectively) were also achieved for the limit of detection and the limit of quantification. The proposed protocol represents a rapid, simple, selective, and sensitive tool to quantify sarcosine in urine samples for prostate cancer diagnosis and for a screening test.


Sarcosine Prostate cancer Alkyl chloroformate Gas chromatography Solid-phase microextraction Tandem mass spectrometry 


  1. 1.
    Sreekumar A, Poisson LM, Rajendiran TM, Khan AP, Cao Q, Yu J et al (2009) Nature 457:910–914CrossRefGoogle Scholar
  2. 2.
    Jamaspishvili T, Kral M, Khomeriki I, Student V, Kolar Bouchal Z (2010) Prostate Cancer Prostatic Dis 13:12–19CrossRefGoogle Scholar
  3. 3.
    Smith JT (1999) Electrophoresis 20:3078–3083CrossRefGoogle Scholar
  4. 4.
    Oguri S, Yokoi K, Motohase Y (1997) J Chromatogr A 787:253–260CrossRefGoogle Scholar
  5. 5.
    Kolch W, Neusüß C, Pelzing M, Mischak H (2005) Mass Spectrom Rev 24:959–977CrossRefGoogle Scholar
  6. 6.
    Williams BJ, Cameron CJ, Workman R, Broeckling CD, Sumner LW, Smith JT (2007) Electrophoresis 28:1371–1379CrossRefGoogle Scholar
  7. 7.
    Le Boucher J, Charret C, Coudray-Lucas C, Giboudeau J, Cynober L (1997) Clin Chem 43:1421–1428Google Scholar
  8. 8.
    Feste AS (1992) J Chromatogr B Biomed Appl 574:23–34CrossRefGoogle Scholar
  9. 9.
    Biggs HG, Gentilcore LJ (1984) Clin Chem 30:851–855Google Scholar
  10. 10.
    Gatti R, Gioia MG (2008) Biomed Chromatogr 22:207–213CrossRefGoogle Scholar
  11. 11.
    Qu J, Wang Y, Luo G, Wu Z, Yang C (2002) Anal Chem 74:2034–2040CrossRefGoogle Scholar
  12. 12.
    Piraud M, Vianey-Saban C, Petritis K, Elfakir C, Steghens JP, Morla A, Bouchu D (2003) Rapid Commun Mass Spectrom 17:1297–1311CrossRefGoogle Scholar
  13. 13.
    Rashed MS, Bucknall MP, Little D, Awad A, Jacob M, Alamoudi M, Alwattar M, Ozand PT (1997) Clin Chem 43:1129–1141Google Scholar
  14. 14.
    Casetta B, Tagliacozzi D, Shushan B, Federici G (2000) Clin Chem Lab Med 38:391–401CrossRefGoogle Scholar
  15. 15.
    Chace DH, Hillman SL, Millington DS, Kahler SG, Roe CR, Naylor EW (1995) Clin Chem 41:62–68Google Scholar
  16. 16.
    Matsumoto I, Kuhara T (1996) Mass Spectrom Rev 15:43–57CrossRefGoogle Scholar
  17. 17.
    Jiang Y, Cheng X, Wang C, Ma Y (2010) Anal Chem 82:9022–9027CrossRefGoogle Scholar
  18. 18.
    Harper JD, Charipar NA, Mulligan CC, Zhang X, Cooks RG, Ouyang Z (2008) Anal Chem 80:9097–9104CrossRefGoogle Scholar
  19. 19.
    García-Reyes JF, Mazzotti F, Harper JD, Charipar NA, Oradu S, Ouyang Z, Sindona G, Cooks RG (2009) Rapid Commun Mass Spectrom 23:3057–3062CrossRefGoogle Scholar
  20. 20.
    Gehrke CW, Nakamoto H, Zumwalt RW (1969) J Chromatogr A 45:24–51CrossRefGoogle Scholar
  21. 21.
    Duncan MW, Poljak A (1998) Anal Chem 70:890–896CrossRefGoogle Scholar
  22. 22.
    Woo KL, Lee DS (1995) J Chromatogr B Biomed Appl 665:15–25CrossRefGoogle Scholar
  23. 23.
    Shen X, Deng C, Wang B, Dong L (2006) Anal Bioanal Chem 384:931–938CrossRefGoogle Scholar
  24. 24.
    Simek P, Heydova A, Jegorov A (1994) J High Resolut Chromatogr 17:145–152CrossRefGoogle Scholar
  25. 25.
    Chaves das Neves HJ, Vasconcelos AMP (1987) J Chromatogr 392:249–258CrossRefGoogle Scholar
  26. 26.
    Starke I, Kleinpeter E, Kamm B (2001) Anal Bioanal Chem 371:380–384Google Scholar
  27. 27.
    Pätzold R, Brückner H (2006) Amino Acids 31:63–72CrossRefGoogle Scholar
  28. 28.
    Davis BA, Durden DA (1987) Biomed Environ Mass Spectrom 14:197–206CrossRefGoogle Scholar
  29. 29.
    Zumwalt RW, Roach D, Gehrke CW (1970) J Chromatogr 53:171–194CrossRefGoogle Scholar
  30. 30.
    Fiamegos YC, Stalikas CD (2006) J Chromatogr A 1110:66–72CrossRefGoogle Scholar
  31. 31.
    Blount BC, Duncan MW (1997) Anal Biochem 244:270–276CrossRefGoogle Scholar
  32. 32.
    Guidetti P, Schwarcz R (2003) Mol Brain Res 118:132–139CrossRefGoogle Scholar
  33. 33.
    Hušek P (1998) J Chromatogr B 717:57–91CrossRefGoogle Scholar
  34. 34.
    Sobolevsky TG, Revelsky AI, Miller B, Oriedo V, Chernetsova ES, Revelsky IA (2003) J Sep Sci 26:1474–1478CrossRefGoogle Scholar
  35. 35.
    Sobolevsky TG, Revelsky AI, Revelsky IA, Miller B, Oriedo V (2004) J Chromatogr B Anal Technol Biomed Life Sci 800:101–107CrossRefGoogle Scholar
  36. 36.
    Huang ZH, Wang J, Gage DA, Watson JT, Sweeley CC, Husek P (2003) J Chromatogr 635:271–281CrossRefGoogle Scholar
  37. 37.
    Haberhauer-Troyer C, Álvarez-Llamas G, Zitting E, Rodríguez-González P, Rosenberg E, Sanz-Medel A (2003) J Chromatogr A 1015:1–10CrossRefGoogle Scholar
  38. 38.
    Vincenti M, Minero C, Lago S, Rovida C (1995) J High Resolut Chromaogr 18:359–362CrossRefGoogle Scholar
  39. 39.
    Husek P (1995) J Chromatogr B Biomed Appl 669:352–357CrossRefGoogle Scholar
  40. 40.
    Namera A, Yashiki M, Nishida M, Kojima T (2002) J Chromatogr B Anal Technol Biomed Life Sci 776:49–55CrossRefGoogle Scholar
  41. 41.
    Villas-Bôas SG, Delicado DG, Åkesson M, Nielsen J (2003) Anal Biochem 322:134–138CrossRefGoogle Scholar
  42. 42.
    Tao X, Liu Y, Wang Y, Qiu Y, Lin J, Zhao A, Su M, Jia W (2008) Anal Bioanal Chem 391:2881–2889CrossRefGoogle Scholar
  43. 43.
    Kaspar H, Dettmer K, Gronwald W, Oefner PJ (2008) J Chromatogr B 870:222–232CrossRefGoogle Scholar
  44. 44.
    Vonderheide AP, Montes-Bayon M, Caruso JA (2002) Analyst 127:49–53CrossRefGoogle Scholar
  45. 45.
    Deng C, Li N, Zhang X (2004) Rapid Commun Mass Spectrom 18:2558–2564CrossRefGoogle Scholar
  46. 46.
    Myung SW, Kim M, Min HK, Yoo EA, Kim KR (1999) J Chromatogr B 727:1–8CrossRefGoogle Scholar
  47. 47.
    Arthur CL, Pawliszyn J (1990) Anal Chem 62:2145–2148CrossRefGoogle Scholar
  48. 48.
    Arthur CL, Killam LM, Buchholz KD, Pawliszyn J, Berg JR (1992) Anal Chem 64:1960–1966CrossRefGoogle Scholar
  49. 49.
    Jentzmik F, Stephan C, Miller K, Schrader M, Erbersdobler A, Kristiansen G, Lein M, Jung K (2010) Eur Urol 58:12–18CrossRefGoogle Scholar
  50. 50.
    Pavlou M, Diamandis EP (2009) Clin Chem 55:1277–1279CrossRefGoogle Scholar
  51. 51.
    Constantinou J, Feneley MR (2006) Prostate Cancer Prostatic Dis 9:6–13CrossRefGoogle Scholar
  52. 52.
    Lord H, Pawliszyn J (2000) J Chromatogr A 885:153–193CrossRefGoogle Scholar
  53. 53.
    Llompard M, Li K, Fingas M (1998) Anal Chem 70:2510–2515CrossRefGoogle Scholar
  54. 54.
    Kaspar H, Dettmer K, Chan Q, Daniels S, Nimkar S, Daviglus ML, Stamler J, Elliott P, Oefner PJ (2009) J Chromatogr B 877:1838–1846CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Brunella Cavaliere
    • 1
  • Barbara Macchione
    • 1
  • Marcello Monteleone
    • 1
  • Attilio Naccarato
    • 1
  • Giovanni Sindona
    • 1
  • Antonio Tagarelli
    • 1
  1. 1.Dipartimento di ChimicaUniversità della CalabriaArcavacata di RendeItaly

Personalised recommendations