Recycling HPLC of p-Bromophenacyl Esters of Saturated C_35–56 Fatty Acids from Mycobacterium Tuberculosis H37Ra on a Silica Column

Kuni Takayama Tuberculosis Research Laboratory, William S. Middleton Memorial Veterans Medical Center, Madison, Wisconsin, 53705; Institute for Enzyme Research University of Wisconsin, 53706

Nilofer Qureshi Tuberculosis Research Laboratory, William S. Middleton Memorial Veterans Medical Center, Madison, Wisconsin, 53705; Institute for Enzyme Research University of Wisconsin, 53706

Howard C. Jordi Application Laboratory Chromatography Supplies Division, Waters Associates, Inc., Milford, Massachusetts, 01757

Heinrich K. Schnoes Department of Biochemistry, College of Agricultural and Life Sciences University of Wisconsin, Madison, Wisconsin, 53706

Abstract

The p-bromophenacyl esters of saturated C_35–56 fatty acids from Mycobacterium tuberculosis H37Ra were separated according to structural classes on a silica column by high performance liquid chromatography (HPLC). The sample was cycled five times during HPLC. Highly purified C_35–38 esters were obtained by this method. Further HPLC fractionation on a reverse-phase column (C₁₈-bonded silica) gave complete separation of most of the remaining fractions. By combining mass spectrometry with HPLC separations, many of the fatty acid esters were tentatively identified.

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Recycling HPLC of p-Bromophenacyl Esters of Saturated C_35–56 Fatty Acids from Mycobacterium Tuberculosis H37Ra on a Silica Column

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Recycling HPLC of p-Bromophenacyl Esters of Saturated C35–56 Fatty Acids from Mycobacterium Tuberculosis H37Ra on a Silica Column

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Recycling HPLC of p-Bromophenacyl Esters of Saturated C_35–56 Fatty Acids from Mycobacterium Tuberculosis H37Ra on a Silica Column