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After incubation for 15 min at 30 C, 1,000 rpm using a 3 mm shaking diameter, cell debris was removed by centrifugation (4,400 g at 4 C for 30 min) and the supernatants were analysed by HPLCMS at two different dilutions (undiluted and four times diluted with 67% methanol, 0.2% hydrochloric acid in H 2 O)

doi:10.1152/physrev.00034.2006 9

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