Abstract
An effective HPLC method is established for the separation of muramic acid, diaminopimelic acid, glutamic acid, and alanine from a bacterial cell wall hydrolysate. In addition the 3-deoxy- d- manno-2-octulosonic acid (KDO) component of outer membrane lipopolysaccharide is quantitatively analyzed. The Bio-Rad HPX-87C carbohydrate and amino acid analysis column is used to separate the cell wall components of Psuedomonas putida under a variety of conditions and resolution of compounds may be modified by altering the calcium ion concentration, temperature, and pH. Optimal conditions for the resolution of most cell wall components are obtained by the use of 50 m m calcium nitrate, pH 3.6, for the mobile phase, at a flow rate of 1 ml per min and at a column temperature of 85 °C. Quantification of compounds may be determined in the minimum of range of 1–2 μg, using a differential refractometer.
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