Chromatographia

, Volume 72, Issue 7–8, pp 641–649 | Cite as

LC-Fluorescence Detection Analysis of Amino Acids from Stellera chamaejasme L. Using 2-[2-(Dibenzocarbazole)-ethoxy] Ethyl Chloroformate as Labeling Reagent

Original

Abstract

A pre-column derivatization method for the sensitive determination of amino acids using the tagging reagent 2-[2-(dibenzocarbazole)-ethoxy] ethyl chloroformate (DBCEC) followed by liquid chromatography with fluorescence detection has been developed. Identification of DBCEC-amino acids derivatives was by liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS–MS). DBCEC can easily and quickly label amino acids, and derivatives are stable enough to be efficiently analyzed by LC. Separation of the derivatized amino acids had been optimized on Hypersil BDS C18 column. A perfect baseline separation for 20 amino acid derivatives was achieved with a ternary gradient elution program. The chromophore of dibenzocarbazole group, which comprise a large rigid planar structure with p–π conjugation system, resulted in a sensitive fluorescence detection for amino acid derivatives. The derivatized amino acids were detected with fluorescence detector with excitation maximum and emission maximum at 300 and 390 nm, respectively. Excellent linear responses were observed with coefficients of >0.9993, and detection limits were in the range of 0.78–5.13 fmol (signal-to-noise ratio of 3). The mean accuracy ranged from 83.4 to 98.7% for fluorescence detection. The mean inter-day precision for all standards was <4.2% of the expected concentration. Therefore, the proposed method was a highly sensitive and specific method for the quantitative analysis of amino acids from biological and natural environmental samples.

Keywords

Column liquid chromatography Fluorescence detection Electrospray mass spectrometry Amino acids Stellera chamaejasme L. 

Notes

Acknowledgments

This work was supported by the National Science Foundation of China (No. 20075016) and supported by 100 Talents Program of the Chinese Academy of Sciences (No. 328).

References

  1. 1.
    Jiang ZH, Tanaka T, Sakamoto T, Kouno I, Duan JA, Zhou RH (2002) Chem Pharm Bull 50:137–139. doi: 10.1248/cpb.50.137 CrossRefGoogle Scholar
  2. 2.
    Yoshida M, Feng WJ, Saijo N, Ikekawa T (1996) Int J Cancer 66:268–270. doi: 10.1002/(SICI)1097-0215(19960410)66:2<268:AID-IJC22>3.0.CO;2-7 CrossRefGoogle Scholar
  3. 3.
    Fan J (2007) J Clin Res 24:2001–2003. doi: CNKI:SUN:HNYZ.0.2007-11-075 Google Scholar
  4. 4.
    Zheng B, Hu H (2006) J Northeast For Univ 34:59–60. doi: cnki:ISSN:1000-5382.0.2006-04-021 (in Chineses)Google Scholar
  5. 5.
    Feng B, Gong X, Li S, Jiang G, Pei Y, Wang Y (2008) Chin J Chin Mater Med 33:403–405. doi: CNKI:SUN:ZGZY.0.2008-04-015 (in Chineses)Google Scholar
  6. 6.
    Feng BM, Pei YH, Hua HM (2004) Chin Chem Lett 15:61–62. doi: cnki:ISSN:1001-8417.0.2004-01-020 Google Scholar
  7. 7.
    Gatti R, Gioia MG, Andreatta P, Pentassuglia G (2004) J Pharm Biomed Anal 35(2):339–348. doi: 10.1016/S0731-7085(03)00584-3 CrossRefGoogle Scholar
  8. 8.
    Dong MW, Gant JR (1985) J Chromatogr A 327:17–25. doi: 10.1016/S0021-9673(01)81635-6 CrossRefGoogle Scholar
  9. 9.
    Hsieh MM, Chen SM (2007) Talanta 73(2):326–331. doi: 10.1016/j.talanta.2007.03.049 CrossRefGoogle Scholar
  10. 10.
    Denoroy L, Parrot S, Renaud L, Renaud B, Zimmer L (2008) J Chromatogr A 1205(1–2):144–149. doi: 10.1016/j.chroma.2008.07.043 CrossRefGoogle Scholar
  11. 11.
    Manica DP, Lapos JA, Jones AD, Ewing AG (2003) Anal Biochem 322(1):68–78. doi: 10.1016/j.ab.2003.07.002 CrossRefGoogle Scholar
  12. 12.
    Aito-Inoue M, Ohtsuki K, NakamurA Y, Park EY, Iwai K, Morimatsu F, Sato K (2006) J Agric Food Chem 54:5261–5266. doi: 10.1021/jf060531s CrossRefGoogle Scholar
  13. 13.
    Poboży E, Czarkowska W, Trojanowicz M (2006) J Biochem Biophys Methods 67(1):37–47. doi: 10.1016/j.jbbm.2006.01.001 CrossRefGoogle Scholar
  14. 14.
    Krause I, Bockhardt A, Neckermann H, Henle T, Klostermeyer H (1995) J Chromatogr A 715:67–79. doi: 10.1016/0021-9673(95)00578-B CrossRefGoogle Scholar
  15. 15.
    Minocha R, Long S (2004) J Chromatogr A 1035:63–73. doi: 10.1016/j.chroma.2004.02.026 CrossRefGoogle Scholar
  16. 16.
    Hanczkó R, Jámbor A, Perl A, Molnár-Perl I (2007) J Chromatogr A 1163:25–42. doi: 10.1016/j.chroma.2007.06.013 CrossRefGoogle Scholar
  17. 17.
    Pereira V, Pontes M, Câmara JS, Marques JC (2008) J Chromatogr A 1189:435–443. doi: 10.1016/j.chroma.2007.12.014 CrossRefGoogle Scholar
  18. 18.
    Cohen SA, Michaud DP (1993) Anal Biochem 211(2):279–287. doi: 10.1006/abio.1993.1270 CrossRefGoogle Scholar
  19. 19.
    Bosch L, Alegría A, Farré R (2006) J Chromatogr B 831:176–183. doi: 10.1016/j.jchromb.2005.12.002 CrossRefGoogle Scholar
  20. 20.
    Lozanov V, Benkova B, Mateva L, Petrov S, Popov E, Slavov C, Mitev V (2007) J Chromatogr B 860:92–97. doi: 10.1016/j.jchromb.2007.10.020 CrossRefGoogle Scholar
  21. 21.
    López-Cervantes J, Sánchez-Machado DI, Rosas-Rodríguez JA (2006) J Chromatogr A 1105:106–110. doi: 10.1016/j.chroma.2005.08.040 CrossRefGoogle Scholar
  22. 22.
    Kőrös Á, Varga ZS, Molnár-Perl I (2008) J Chromatogr A 1203:146–152. doi: 10.1016/j.chroma.2008.07.035 CrossRefGoogle Scholar
  23. 23.
    You J, Shan Y, Zhen L, Zhang L, Zhang Y (2003) Anal Biochem 313(1):17–27. doi: 10.1016/S0003-2697(02)00398-6 CrossRefGoogle Scholar
  24. 24.
    You J, Ming Y, Shi Y, Zhao X, Suo Y, Wang H, Li Y, Sun J (2005) Talanta 68(2):448–458. doi: 10.1016/j.talanta.2005.09.019 CrossRefGoogle Scholar
  25. 25.
    You J, Liu L, Zhao W, Zhao X, Suo Y, Wang H, Li Y (2007) Anal Bioanal Chem 387(8):2705–2718. doi: 10.1007/s00216-007-1155-9 CrossRefGoogle Scholar
  26. 26.
    You J, Song C, Yan T, Sun Z, Li Y, Suo Y (2010) Anal Chim Acta 658(1):98–105. doi: 10.1016/j.aca.2009.10.061 CrossRefGoogle Scholar
  27. 27.
    Montigny PD, Stobaugh JF, Givens RS, Carlson RG, Srinivasachar K, Sternson LA, Higuchi T (1987) Anal Chem 59(8):1096–1101. doi: 10.1021/ac00135a007 CrossRefGoogle Scholar
  28. 28.
    Campanella L, Crescentini G, Avino P (1999) J Chromatogr A 833:137–145. doi: 10.1016/S0021-9673(98)01023-1 CrossRefGoogle Scholar

Copyright information

© Vieweg+Teubner Verlag | Springer Fachmedien Wiesbaden GmbH 2010

Authors and Affiliations

  1. 1.Northwest Plateau Institute of BiologyChinese Academy of ScienceXiningPeople’s Republic of China
  2. 2.Shandong Provincial Key Laboratory of Life-Organic AnalysisQufu Normal UniversityQufuPeople’s Republic of China
  3. 3.Graduate University of the Chinese Academy of ScienceBeijingPeople’s Republic of China

Personalised recommendations