I am trying to set up a reaction by incubating linear isoprenoid precursor Farnesyl diphosphate with the recombinant enzyme in 50mM Tris-Cl (pH 8.2) buffer containing 20% v/v glycerol, 2 mM MgCl2 and 0.2 mM 2-mercaptoethanol for 4 hours at 30oC, overlaid with HPLC grade pentane. Extraction to be done thrice with pentane: CH2Cl2 (5:1). The combined organic layers to be dried by passage through MgSO4 followed by Na2SO4. Further, concentration under reduced pressure at 0oC to 200 UL.
The expected product is a highly volatile bicyclic terpenoid.
Q1. Do the extraction and drying procedures, mentioned above, require to be conducted at 0-4oC?
Q2. What are the other ways to extract, dry and concentrate such volatiles?
The problem of isolating volatile components after extraction is that they start to evaporate rapidly as soon as the solvent is gone. One trick is that is used by the analysis of volatile polycyclic aromatics is to add a small amount of non- or less volatile cosolvent to the combined extracts, for example 1 mL (or less if you like) of hexadecane. After the very volatile pentane and dichloromethane solvents are evaporated your product is concentrated in the cosolvent. Check before if the selected cosolvent might interfere with further analysis, for example check if the chromatographic elution times are sufficiently different.
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