Immunoaffinity chromatography of polycyclic aromatic hydrocarbons in columns prepared by the sol-gel method

Abstract This paper describes the immobilisation of the IgG fraction of an anti-pyrene antiserum by the sol-gel method and evaluates the performance of the immunoaffinity columns prepared with respect to column stability, capacity, selectivity and non-specific binding on the glass matrix. The immunoaffinity columns prepared had a pyrene-breakthrough capacity of 78 ng g −1 gel and 38 ng mg −1 IgG. Pyrene could be isolated and enriched by a factor of 125. No change in specific retention properties could be observed after 10 absorption/desorption cycles. Cross-reactivity of the antibodies with some of the polycyclic aromatic hydrocarbons (PAHs) selected by the US EPA and German Authorities (TVO standards) to monitor PAH contamination resulted in quantitative retention of fluoranthene and some retention of fluorene, phenanthrene and anthracene. Non-specific adsorption on the glass matrix dominates the retention of the TVO- and EPA-PAHs with log K ow >5.2. Storage of the columns in buffer slowly increases non-specific binding.