n the present work, a proof of principle clean up method was developed, aiming for a maximum recovery of PDIMs and minimum collection of cholesterol from a stock solution of PDIMs and cholesterol standards in PE. The lipid extraction from sputum samples was performed with a combination of apolar petroleum ether (PE) and an immiscible polar solvent (methanol), applying a modified Dobson protocol [10]. Four PE extracts of positive sputum samples were selected that were previously [15] found to have high amounts of cholesterol. These were then passed through different solid phase extraction (SPE) materials, both commercial and molecularly imprinted polymers (MIPs) cartridges. Molecular imprinting is performed by producing a polymeric matrix, containing complementary residues, in the presence of the target molecule [16], in this case cholesterol. After formation, the original template is removed, leaving cavities that are complementary to the shape and chemical profile of the template, only allowing specific recognition and rebinding.