3.2.1. Reaction Conditions

1. Place appropriate amounts of glycerol, CLA, and molecular sieves in a flask. Typically, 2.0 g CLA, 0.8 g molecular sieves, and sufficient glycerol to provide the desired molar ratio of CLA to glycerol are added to a 25-mL flask (see Notes 12-15).

2. Preheat the mixture to the desired temperature.

3. Place the flask containing the reactants in an orbital shaker maintained at the desired (constant) temperature and add the biocatalyst (0.3 g Lipozyme IM, for the quantities of material specified in step 1 (see Note 16). Depending on the stoichiometric ratio of reactants, the reaction conditions employed, and the time for reaction, one can produce primarily mono-, di-, or triacylglycerols or mixtures thereof (cf. Fig. 3).

4. Analyze the reaction mixture by HPLC (see Note 17).

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Fig. 3. Time-course of the distribution of reactant and product species in the esteri-fication of glycerol with CLA. Conditions: 2000 mg CLA, 217 mg glycerol, 300 mg Lipozyme IM, 800 molecular sieves, 50°C.

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