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Detection of Amine Impurity and Quality Assessment of the MALDI Matrix α-Cyano-4-Hydroxy-Cinnamic Acid for Peptide Analysis in the amol Range

  • Focus: Development and Application of TOF and TOF/TOF MS: Research Article
  • Published:
Journal of The American Society for Mass Spectrometry

Abstract

We have studied sample preparation conditions to increase the reproducibility of positive UV-MALDI-TOF mass spectrometry of peptides in the amol range. By evaluating several α-cyano-4-hydroxy-cinnamic acid (CHCA) matrix batches and preparation protocols, it became apparent that two factors have a large influence on the reproducibility and the quality of the generated peptide mass spectra: (1) the selection of the CHCA matrix, which allows the most sensitive measurements and an easier finding of the “sweet spots,” and (2) the amount of the sample volume deposited onto the thin crystalline matrix layer. We have studied in detail the influence of a contaminant, coming from commercial CHCA matrix batches, on sensitivity of generated peptide mass spectra in the amol as well as fmol range of a tryptic peptide mixture. The structure of the contaminant, N,N-dimethylbutyl amine, was determined by applying MALDI-FT-ICR mass spectrometry experiments for elemental composition and MALDI high energy CID experiments utilizing a tandem mass spectrometer (TOF/RTOF). A recrystallization of heavily contaminated CHCA batches that reduces or eliminates the determined impurity is described. Furthermore, a fast and reliable method for the assessment of CHCA matrix batches prior to tryptic peptide MALDI mass spectrometric analyses is presented.

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References

  1. Karas, M., Bachmann, D., Bahr, U., Hillenkamp, F.: Matrix-assisted ultraviolet laser desorption of non-volatile compounds. Int. J. Mass Spectrom. 78, 53–56 (1987)

    Article  CAS  Google Scholar 

  2. Tanaka, K., Waki, H., Ido, Y., Akita, S., Yoshida, Y.: Protein and polymer analysis up to m/z 100,000 by laser ionization time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 2, 151–153 (1988)

    Article  CAS  Google Scholar 

  3. Vestal, M.J., Juhasz, P., Martin, S.A.: Delayed extraction matrix-assisted laser desorption time-of flight mass spectrometry. Rapid Commun. Mass Spectrom. 9, 1044–1050 (1995)

    Article  CAS  Google Scholar 

  4. Kaufmann, R., Chaurand, P., Kirsch, D., Spengler, B.: Post-source decay and delayed extraction in matrix-assisted laser desorption/ionization-reflectron time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 10, 1199–1208 (1996)

    Article  CAS  Google Scholar 

  5. Kussmann, M., Nordhoff, E., Rahbek-Nielsen, H., Haebel, S., Rossel-Larsen, M., Jakobsen, L., Gobom, J., Mirgorodskaya, E., Kroll-Kirstensen, A., Palm, L., Roepstorff, P.: Matrix-assisted laser desorption/ionization mass spectrometry sample preparation techniques designed for various peptide and protein analytes. J. Mass Spectrom. 32, 593–601 (1997)

    Article  CAS  Google Scholar 

  6. Landry, F., Lombardo, C.H., Smith, J.W.: A method for application of samples to matrix-assisted laser desorption ionization time-of-flight targets that enhances peptide detection. Anal. Biochem. 279, 1–8 (2000)

    Article  CAS  Google Scholar 

  7. Keller, B.O., Li, L.: Detection of 2500 molecules of substance P by MALDI TOF mass spectrometry and investigations into the fundamental limits of detection in MALDI. J. Am. Soc. Mass Spectrom. 12, 1055–1063 (2001)

    Article  CAS  Google Scholar 

  8. Vorm, O., Roepstorff, P., Mann, M.: Improved resolution and very high sensitivity in MALDI time-of-flight surfaces made by fast evaporation. Anal. Chem. 66, 3281–3287 (1994)

    Article  CAS  Google Scholar 

  9. Lake, D.A., Johnson, M.V., McEwen, C.N., Larsen, B.S.: Sample preparation for high throughput accurate mass analysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Rapid Commun. Mass Spectrom. 14, 1008–1013 (2000)

    Article  CAS  Google Scholar 

  10. Beavis, R.C., Chaudhary, T., Chait, B.T.: α-Cyano-4-hydroxy cinnamic acid as a matrix for matrix-assisted laser desorption mass spectrometry. Org. Mass Spectrom. 27, 156–158 (1992)

    Google Scholar 

  11. Gobom, J., Schuerenberg, M., Mueller, M., Theiss, D., Lehrach, H., Nordhoff, E.: α-Cyano-4-hydroxycinnamic acid affinity sample preparation. A protocol for MALDI MS peptide analysis in proteomics. Anal. Chem. 73, 434–438 (2001)

    Google Scholar 

  12. Vestal, M., Juhasz, P., Hines, W., Martin, S.: A new delayed extraction MALDI-TOF MS-MS for characterization of protein digests. In: Burlingame, A.L., Carr, S.A., Baldwin, M.A. (eds.) Mass spectrometry in biology and medicine, pp. 1–16. Humana Press, New York (1999)

    Google Scholar 

  13. Schaub, T.M., Hendrickson, C.L., Quinn, J.P., Rodgers, R.P., Marshall, A.G.: Instrumentation and method for ultrahigh resolution field desorption ionization Fourier transform ion cyclotron resonance mass spectrometry of nonpolar species. Anal. Chem. 77, 1317–1324 (2005)

    Article  CAS  Google Scholar 

  14. Senko, M.W., Canterbury, J.D., Guan, S., Marshall, A.G.: A high-performance modular data system for Fourier transform ion cyclotron resonance mass spectrometry. Rapid Commun. Mass Spectrom. 10, 1839–1844 (1996)

    Article  CAS  Google Scholar 

  15. Blakney, G.T., Hendrickson, C.L., Marshall, A.G.: Predator data station: A fast data acquisition system for advanced FT-ICR MS experiments. Int. J. Mass Spectrom. 306, 246–252 (2011)

    Article  CAS  Google Scholar 

  16. Marshall, A.G., Roe, D.C.: Theory of Fourier transform ion cyclotron mass spectrometry: response to frequency-sweep excitation. J. Chem. Phys. 73, 1581–1590 (1980)

    Article  CAS  Google Scholar 

  17. Li, L., Golding, R.E., Whittal, R.M.: Detection of 25,000 molecules of Substance P by MALDI-TOF mass spectrometry and investigations into the fundamental limits of the detection. J. Am. Chem. Soc. 188, 11662–11663 (2001)

    Google Scholar 

  18. Zhang, H., Andren, P.E., Caprioli, R.M.: Micro-preparation procedure for high-sensitivity matrix-assisted laser desorption ionization mass spectrometry. J. Mass Spectrom. 30, 1768–1771 (1995)

    Article  CAS  Google Scholar 

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Acknowledgments

The authors acknowledge support for this work by the Austrian Science Foundation (grant 14181 to G.A.), and support in part by the National Science Foundation (CHE-99-09502), Florida State University, Ohio State University, and the National High Magnetic Field Laboratory in Tallahassee, FL, USA. The authors appreciate the instrumental support of Alan G. Marshall and Christopher L. Hendrickson (both Tallahassee, FL, USA). In the meantime high-quality CHCA matrix batches can purchased from some vendors. Furthermore, the authors thank K. Jackson (Manchester, UK) and M. Karas (Frankfurt/Main, Germany) for the helpful discussions during the characterization of the CHCA contaminant.

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Author notes

  1. Tanner M. Schaub

    Present address: Chemical Analysis and Instrumentation Laboratory, New Mexico State University, Las Cruces, NM, USA

Authors and Affiliations

  1. Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria

    Justyna Rechthaler, Ernst Pittenauer & Günter Allmaier

  2. Department of Chemistry, Florida State University, Tallahassee, FL, USA

    Tanner M. Schaub

Authors
  1. Justyna Rechthaler
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  2. Ernst Pittenauer
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  3. Tanner M. Schaub
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  4. Günter Allmaier
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Corresponding author

Correspondence to Günter Allmaier.

Additional information

This paper is dedicated to our colleague Professor Bob Cotter in recognition of the 2011 ASMS Award for Distinguished Contribution to Mass Spectrometry

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Rechthaler, J., Pittenauer, E., Schaub, T.M. et al. Detection of Amine Impurity and Quality Assessment of the MALDI Matrix α-Cyano-4-Hydroxy-Cinnamic Acid for Peptide Analysis in the amol Range. J. Am. Soc. Mass Spectrom. 24, 701–710 (2013). https://doi.org/10.1007/s13361-013-0614-0

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  • DOI: https://doi.org/10.1007/s13361-013-0614-0

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