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The use of solid-phase filtration column (MycoSep 228®) and liquid chromatography for determination of patulin in apple and hawthorn beverages was investigated. The method passed a series of validation tests, including separation efficiency, sensitivity, repeatability, validation within a collaborative trial, recoveries, and applicability. Accuracy and precision were very good for determination of patulin in both spiked and naturally contaminated apple/hawthorn beverage samples at levels of 10 μg/L and above, with limits of detection and quantification at 8 and 23 μg/L, respectively. The applicability of the method was tested by analyzing commercially available domestic apple and hawthorn beverages. Patulin was found in 3 of 52 (5.77%) apple beverages at concentrations ranging from 63.45 to 88.44 μg/L, and in 6 of 43 (13.95%) hawthorn beverages ranging from 19.8 to 206.88 μg/L. Among these were 3 apple and 4 hawthorn beverage samples with toxin levels that exceeded the Chinese legislative requirement of 50 μg/L in apple- and hawthorn-based products intended for human consumption. Comparable tests revealed that the results obtained by both the new MycoSep-based method and the official AOAC method are not significantly different (t value 0.003268 < t0.05, P > 0.05).

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To obtain high titer monoclonal antibodies (McAbs) which can react with mammalian prion protein (PrP), Balb/C mice were immunized with bovine (Bo) PrP peptide (BoPrP 209-228 aa) coupled to keyhole limpet hemocyanin (KLH). The hybridoma cell lines secreting monoclonal antibodies against the peptide were established by cell fusion and cloning. The obtained McAbs were applied to detect recombinant human, bovine and hamster PrP, cellular prion protein (PrP(c)) in normal bovine brain and pathogenic scrapie prion protein (PrP(Sc)) accumulated in the medulla oblongata of bovine spongiform encephalopathy(BSE)specimen with Western blot and immunohistochemical detection, respectively. The current procedure might offer a simple, feasible method to raise high titer antibodies for studying biological features of PrP in mammals, as well as detection of transmissible spongiform encephalopathy (TSE) and diagnosis of BSE, in particular.

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