Semi-preparative high-performance liquid chromatography has been used for the preparation of copper(II) bacteriochlorophyll a [Cu(II)-BCh]-a] and zinc(II) bacteriochlorophyll a [Zn(II)-BCh]-a]. Both compounds are separated on a reversed-phase Inertsil ODS-2 column using a mobile phase of acetone-methanol (25:75, v/v). The fractionated metallobacteriochlorophylls (M-Bchl-a) are identified by fast atom bombardment mass spectrometry. The spectroscopic parameters such as the wavelength of absorption maxima and the molar extinction coefficients are determined using pure M-Bchl-a obtained by preparative HPLC. The HPLC method proposed here has been demonstrated to be useful for the purification and determination of components of M-Bchl-a.
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