A small portion of Spat
was analyzed by HPLC to confirm that patulin disappeared
completely. The cells were washed with phosphate buffer
(50 mM, pH 7.0) three times. Three grams of wet cells were quickly
ground with quartz sand and liquid nitrogen and suspended in
7 ml phosphate buffer. The samples were centrifuged at 4 C at
13,000g for 10 min and the supernatant was collected. The intracellular
protein solutions made from Sample 1 and Sample 2 were
marked as I and I-pat, respectively. Fifteen micrograms of patulin
was added to 5 ml S, S-pat, I, and I-pat at 25 C and the patulin concentrations
were determined after 24 h.