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More recently a four-layer gradient has been described for fractionating Paracentrous lividus: OptiPrepTM was diluted with 0.5M NaCl, 10 mM EDTA to produce four solutions of 10%, 20% 30% and 70% (v/v) OptiPrep (equivalent to 6%, 12%, 18% and 42% w/v iodixanol)

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Absorbance was measured at 227nm and drug content was calculated