, Volume 33, Issue 11–12, pp 567–570 | Cite as

Preparation and determination of manganese(III) chlorophylls by reversed-phase high performance liquid chromatography

  • S. Li
  • H. Inoue


Manganese chlorophylls have been synthesized by refluxing a mixture of pheophytins dissolved in acetone and manganese(II) acetate anhydride dissolved in glacial acetic acid. A good separation of manganese(III) chlorophylls has been attained by RP-HPLC using chemically bonded C18 silica as a stationary phase and methanol with 3% acetic acid as a mobile phase. An accurate and rapid HPLC method is described for the simultaneous determination of manganese(III) chlorophyll-a [Mn(III)-chl-a] and manganese(III) chlorophyll-b [Mn(III)-chl-b]. Tailing arising from dissociation of acetate ions is improved by addition of sodium acetate (5×10−3 M) to the mobile phase (acetone: methanol=90∶10, vol/vol). The analytical values obtained by the HPLC method are very close to the calculated contents in all samples, but those obtained by spectrophotometry are high because of the interferences from overlapping of absorption bands. In the proposed HPLC method the calibration graphs of Mn(III)-chl-a and Mn(III)-chl-b are linear in the concentration range 0–20 μg cm−3 with relative standard deviations (n=10) of 3.46% and 4.54%, respectively.

Key Words

Column liquid chromatography Chlorophylls Manganese(III) chlorophylls 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. [1]
    Y. Yoshitake, K. Kakiuchi, K. Morishige, S. Gohda, T. Shigematsu, Y. Nishikawa, Anal. Sci.,2, 67 (1986).Google Scholar
  2. [2]
    A. P. Murray, C. F. Gibbs, A. R. Longnore, Marine Chemistry,19, 211 (1986).Google Scholar
  3. [3]
    M. Gapata, A. M. Ayala, J. M. Franco, J. L. Garrido, Chromatographia,23, 26 (1987).Google Scholar
  4. [4]
    K. Furuya, H. Inoue, T. Shirai, Anal. Sci.,3, 353 (1987).Google Scholar
  5. [5]
    H. Inoue, K. Furuya, K. Watanabe, K. Tanaka, T. Shirai, E. Miyoshi, Anal. Sci.,4, 599 (1988).Google Scholar
  6. [6]
    K. Furuya, N. Ohki, H. Inoue, T. Shirai, Chromatographia,25, 319 (1988).Google Scholar
  7. [7]
    M. Aizawa, H. Shinohara, S. Watanabe, Denki Kagaku,50, 854 (1982).Google Scholar
  8. [8]
    M. Sato, I. Fujimoto, T. Sakai, R. Kimura, T. Murata, Yakugaku Zasshi,99, 1055 (1979).Google Scholar
  9. [9]
    R. B. Paik, N. G. Pon, J. Mol. Biol.,6, 105 (1963).Google Scholar
  10. [10]
    L. P. Vernon, Anal. Chem.,32, 1144 (1960).Google Scholar
  11. [11]
    Y. Yoshitake, S. Gohda, H. Yamazaki, K. Morishige, Y. Nishikawa, T. Shigematsu, Bunseki Kagaku,33, 667 (1984).Google Scholar
  12. [12]
    Y. Nishikawa, K. Hiraki, Y. Onoue, K. Nishikawa, Y. Yoshitake, T. Shigematsu, Bunseki Kagaku,32, E115 (1983).Google Scholar
  13. [13]
    K. Iriyama, M. Shiraki, M. Yoshiura, J. Liq. Chromatogr.,2, 155 (1979).Google Scholar
  14. [14]
    L. J. Boucher, Coord. Chem. Rev.,7, 289 (1972).Google Scholar
  15. [15]
    S. Li, H. Inoue, Anal. Sci.,7, Suppl., 121 (1991).Google Scholar
  16. [16]
    H. Inoue, M. Imai, T. Shirai, The 47th Symposium on Anal. Chem., 107 (1986).Google Scholar

Copyright information

© Friedr. Vieweg & Sohn Verlagsgesellschaft mbH 1992

Authors and Affiliations

  • S. Li
    • 1
    • 2
  • H. Inoue
    • 1
  1. 1.Department of Applied Chemistry, Faculty of Science and TechnologyKeio UniversityYokohamaJapan
  2. 2.Department of Technological PhysicsPeking UniversityBeijingP.R. China

Personalised recommendations