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Structural Characterization of Chlorophyll-a by High Resolution Tandem Mass Spectrometry

  • Research Article
  • Published:
Journal of The American Society for Mass Spectrometry

Abstract

A high resolution Fourier transform ion cyclotron resonance (FTICR) mass spectrometer is used for characterizing the fragmentation of chlorophyll-a. Three tandem mass spectrometry (MS/MS) techniques, including electron-induced dissociation (EID), collisionally activated dissociation (CAD), and infrared mutiphoton dissociation (IRMPD) are applied to the singly protonated chlorophyll-a. Some previously unpublished fragments are identified unambiguously by utilizing high resolution and accurate mass value provided by the FTICR mass spectrometer. According to this research, the two long aliphatic side chains are shown to be the most labile parts, and favorable cleavage sites are proposed. Even though similar fragmentation patterns are generated by all three methods, there are much more abundant peaks in EID and IRMPD spectra. The similarities and differences are discussed in detail. Comparatively, cleavage leading to odd electron species and H• loss both seem more common in EID experiments. Extensive loss of small side groups (e.g., methyl and ethyl) next to the macrocyclic ring was observed. Coupling the high performance FTICR mass spectrometer with contemporary MS/MS techniques, especially IRMPD and EID, proved to be very promising for the structural characterization of chlorophyll, which is also suitable for the rapid and accurate structural investigation of other singly charged porphyrinic compounds.

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References

  1. Björn, L., Papageorgiou, G., Blankenship, R., Govindjee, G.: A viewpoint: why chlorophyll-a. Photosynth. Res. 99, 85–98 (2009)

    Article  Google Scholar 

  2. Aronoff, S., Mackinney, G.: The photo-oxidation of chlorophyll. J. Am. Chem. Soc. 65, 956–958 (1943)

    Article  CAS  Google Scholar 

  3. Scheer, H.: In: Grimm, B., Porra, R.J., Rüdiger, W., Scheer, H. (eds.) Chlorophylls and bacteriochlorophylls, pp. 1–26. Springer, The Netherlands (2006)

    Chapter  Google Scholar 

  4. De Paula, J.C., Robblee, J.H., Pasternack, R.F.: Aggregation of chlorophyll-a probed by resonance light scattering spectroscopy. Biophys. J. 68, 335–341 (1995)

    Article  Google Scholar 

  5. Wang, X.-F., Xiang, J., Wang, P., Koyama, Y., Yanagida, S., Wada, Y., Hamada, K., Sasaki, S.-i., Tamiaki, H.: Dye-sensitized solar cells using a chlorophyll-a derivative as the sensitizer and carotenoids having different conjugation lengths as redox spacers. Chem. Phys. Lett. 408, 409–414 (2005)

    Article  CAS  Google Scholar 

  6. Hörtensteiner, S., Kräutler, B.: Chlorophyll breakdown in higher plants. Biochim. Biophys. Acta Bioenerg. 1807, 977–988 (2011)

    Article  Google Scholar 

  7. Sleno, L., Windust, A., Volmer, D.: Structural study of spirolide marine toxins by mass spectrometry. Anal. Bioanal. Chem. 378, 969–976 (2004)

    Article  CAS  Google Scholar 

  8. Mosely, J.A., Smith, M.J.P., Prakash, A.S., Sims, M., Bristow, A.W.T.: Electron-induced dissociation of singly charged organic cations as a tool for structural characterization of pharmaceutical type molecules. Anal. Chem. 83, 4068–4075 (2011)

    Article  CAS  Google Scholar 

  9. Wolff, J.J., Laremore, T.N., Aslam, H., Linhardt, R.J., Amster, I.J.: Electron-induced dissociation of glycosaminoglycan tetrasaccharides. J. Am. Soc. Mass. Spectrom. 19, 1449–1458 (2008)

    Article  CAS  Google Scholar 

  10. Hanson, C.W., Thaler, E.R.: Electronic nose prediction of a clinical pneumonia score: biosensors and microbes. Anesthesiology 102, 63–68 (2005)

    Article  Google Scholar 

  11. Van Breemen, R.B., Canjura, F.L., Schwartz, S.J.: Identification of chlorophyll derivatives by mass spectrometry. J. Agric. Food Chem. 39, 1452–1456 (1991)

    Article  Google Scholar 

  12. Suman, M., De Maria, R., Catellani, D.: Chromatographic evaluation of chlorophyll derivatives in pasta-based food products: effects of pasteurization treatments and correlation with sensory profiles. J. Sci. Food Agric. 88, 471–478 (2008)

    Article  CAS  Google Scholar 

  13. Kao, T.H., Chen, C.J., Chen, B.H.: An improved high performance liquid chromatography-photodiode array detection-atmospheric pressure chemical ionization-mass spectrometry method for determination of chlorophylls and their derivatives in freeze-dried and hot-air-dried Rhinacanthus nasutus (L.) Kurz. Talanta 86, 349–355 (2011)

    Article  CAS  Google Scholar 

  14. Bale, N.J., Llewellyn, C.A., Airs, R.L.: Atmospheric pressure chemical ionisation liquid chromatography/mass spectrometry of type II chlorophyll-a transformation products: diagnostic fragmentation patterns. Org. Geochem. 41, 473–481 (2010)

    Article  CAS  Google Scholar 

  15. Suzuki, T., Midonoya, H., Shioi, Y.: Analysis of chlorophylls and their derivatives by matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry. Anal. Biochem. 390, 57–62 (2009)

    Article  CAS  Google Scholar 

  16. Hunt, J.E., Macfarlane, R.D., Katz, J.J., Dougherty, R.C.: High-energy fragmentation of chlorophyll-a and its fully deuterated analog by californium-252 plasma desorption mass spectrometry. J. Am. Chem. Soc. 103, 6775–6778 (1981)

    Article  CAS  Google Scholar 

  17. Chait, B.T., Field, F.H.: Californium-252 fission fragment ionization mass spectrometry of chlorophyll-a. J. Am. Chem. Soc. 106, 1931–1938 (1984)

    Article  CAS  Google Scholar 

  18. Grese, R.P., Cerny, R.L., Gross, M.L., Senge, M.: Determination of structure and properties of modified chlorophylls by using fast atom bombardment combined with tandem mass spectrometry. J. Am. Soc. Mass. Spectrom. 1, 72–84 (1990)

    Article  CAS  Google Scholar 

  19. Dale, M.J., Costello, K.F., Jones, A.C., Langridge-Smith, P.R.R.: Investigation of porphyrins and metalloporphyrins using two-step laser mass spectrometry. J. Mass Spectrom. 31, 590–601 (1996)

    Article  CAS  Google Scholar 

  20. Van Berkel, G.J., Glish, G.L., McLuckey, S.A., Tuinman, A.A.: Porphyrin pyrrole sequencing: low-energy collision-induced dissociation of (M + 7H)+ generated in-situ during ammonia chemical ionization. Anal. Chem. 62, 786–793 (1990)

    Article  Google Scholar 

  21. Rosario, M., Domingues, M., Nemirovskiy, O., Graço, M., Marques, O., Neves, M., Cavaleiro, J., Ferrer-Correia, A., Gross, M.: High- and low-energy collisionally activated decompositions of octaethylporphyrin and its metal complexes. J. Am. Soc. Mass. Spectrom. 9, 767–774 (1998)

    Article  Google Scholar 

  22. Mayer, P.M., Poon, C.: The mechanisms of collisional activation of ions in mass spectrometry. Mass Spectrom. Rev. 28, 608–639 (2009)

    Article  CAS  Google Scholar 

  23. Zubarev, R.A., Kelleher, N.L., McLafferty, F.W.: Electron capture dissociation of multiply charged protein cations. A nonergodic process. J. Am. Chem. Soc. 120, 3265–3266 (1998)

    Article  CAS  Google Scholar 

  24. Zubarev, R.A., Horn, D.M., Fridriksson, E.K., Kelleher, N.L., Kruger, N.A., Lewis, M.A., Carpenter, B.K., McLafferty, F.W.: Electron capture dissociation for structural characterization of multiply charged protein cations. Anal. Chem. 72, 563–573 (2000)

    Article  CAS  Google Scholar 

  25. Lioe, H., O’Hair, R.: Comparison of collision-induced dissociation and electron-induced dissociation of singly protonated aromatic amino acids, cystine and related simple peptides using a hybrid linear ion trap-FT-ICR mass spectrometer. Anal. Bioanal. Chem. 389, 1429–1437 (2007)

    Article  CAS  Google Scholar 

  26. Khairallah, G.N., O’Hair, R.A.J., Bruce, M.I.: Gas-phase synthesis and reactivity of binuclear gold hydride cations, (R3PAu)2H+ (R = Me and Ph). Dalton Trans. doi:10.1039/B604404B (2006)

  27. Wills, R.H., Tosin, M., O’Connor, P.B.: Structural characterization of polyketides using high mass accuracy tandem mass spectrometry. Anal. Chem. 84, 8863–8870 (2012)

    Article  CAS  Google Scholar 

  28. Kaczorowska, M.A., Cooper, H.J.: Electron induced dissociation (EID) tandem mass spectrometry of octaethylporphyrin and its iron(III) complex. Chem. Commun. 47, 418–420 (2011)

    Article  CAS  Google Scholar 

  29. Bernigaud, V., Drenck, K., Huber, B.A., Hvelplund, P., Jabot, T., Kadhane, U., Kirketerp, M.-B.S., Liu, B., Lykkegaard, M.K., Manil, B., Nielsen, S.B.: Electron capture induceddissociation of protoporphyrin IX ions. J. Am. Soc. Mass. Spectrom. 19, 809–813 (2008)

    Article  CAS  Google Scholar 

  30. Zhao, C., Sethuraman, M., Clavreul, N., Kaur, P., Cohen, R.A., O’Connor, P.B.: Detailed map of oxidative post-translational modifications of human P21Ras using Fourier transform mass spectrometry. Anal. Chem. 78, 5134–5142 (2006)

    Article  CAS  Google Scholar 

  31. Caravatti, P., Allemann, M.: The ‘infinity cell’: a new trapped-ion cell with radiofrequency covered trapping electrodes for fourier transform ion cyclotron resonance mass spectrometry. Org. Mass Spectrom. 26, 514–518 (1991)

    Article  CAS  Google Scholar 

  32. Fiedor, L., Kania, A., Myśliwa-Kurdziel, B., Orzeł, Ł., Stochel, G.: Understanding chlorophylls: central magnesium ion and phytyl as structural determinants. Biochim. Biophys. Acta Bioenerg. 1777, 1491–1500 (2008)

    Article  CAS  Google Scholar 

  33. Qi, Y., Thompson, C., Van Orden, S., O’Connor, P.: Phase correction of Fourier transform ion cyclotron resonance mass spectra using MatLab. J. Am. Soc. Mass. Spectrom. 22, 138–147 (2011)

    Article  CAS  Google Scholar 

  34. de Hoffmann, E., Stroobant, V.: Mass spectrometry: principles and applications. John Wiley and Son Ltd, England (2007)

    Google Scholar 

  35. Chow, H.-C., Serlin, R., Strouse, C.E.: Crystal and molecular structure and absolute configuration of ethyl chlorophyllide a dihydrate. Model for the different spectral forms of chlorophyll a. J. Am. Chem. Soc. 97, 7230–7237 (1975)

    Article  CAS  Google Scholar 

  36. Zubarev, R.A., Haselmann, K.F., Budnik, B., Kjeldsen, F., Jensen, F.: Towards an understanding of the mechanism of electron-capture dissociation: a historical perspective and modern ideas. Eur. J. Mass Spectrom. 8, 337–349 (2002)

    Article  CAS  Google Scholar 

  37. Grese, R.P., Cerny, R.L., Gross, M.L., Senge, M.: Determination of structure and properties of modified chlorophylls by using fast atom bombardment combined with tandem mass spectrometry. J. Am. Soc. Mass. Spectrom. 1, 72–84 (1990)

    Article  CAS  Google Scholar 

  38. Mauzerall, D.: Multiple excitations in photosynthetic systems. Biophys. J. 16, 87–91 (1976)

    Article  CAS  Google Scholar 

  39. Weigl, J.W., Livingston, R.: Infrared spectra of chlorophyll andrelated compounds. J. Am. Chem. Soc. 75, 2173–2176 (1953)

    Article  CAS  Google Scholar 

  40. Holt, A.S., Jacobs, E.E.: Infra-red absorption spectra of chlorophylls and derivatives. Plant Physiol. 30, 553–559 (1955)

    Article  CAS  Google Scholar 

  41. Eckardt, N.A.: A new chlorophyll degradation pathway. Plant Cell 21, 700 (2009)

    Article  CAS  Google Scholar 

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Acknowledgments

The authors acknowledge support for this work by the Chancellor’s International Scholarships of Warwick, Warwick Centre for Analytical Science (EPSRC funded EP/F034210/1), the University of Warwick, and the Department of Chemistry. Special thanks are due to Dr. David Kilgour for his help with the phasing work and discussion. The authors also thank Yulin Qi, Pilar Perez-Hurtado, Rebecca Wills, Andrea Lopez-Clavijo, Tzu-yung Lin, Andrew Soulby, and Samantha Benson for their help.

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Authors and Affiliations

  1. Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK

    Juan Wei, Huilin Li, Mark P. Barrow & Peter B. O’Connor

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  1. Juan Wei
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  2. Huilin Li
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  4. Peter B. O’Connor
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Correspondence to Peter B. O’Connor.

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Wei, J., Li, H., Barrow, M.P. et al. Structural Characterization of Chlorophyll-a by High Resolution Tandem Mass Spectrometry. J. Am. Soc. Mass Spectrom. 24, 753–760 (2013). https://doi.org/10.1007/s13361-013-0577-1

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