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5 mL 11 v/v methanol-acetonitrile followed by 0.5 mL methanol; eluate reduction to 0.25 mL compensated the initial sample dilution. The accuracy was assessed in certified blank fetal bovine serum and in human plasma, gaining satisfactory recovery in the range 65-122%, detection limits in the range 0.02-0.3 ng mL-1 (0.8 ng mL-1 for 17-β-estradiol) and suitable inter-day precision (relative standard deviation (RSD) less then 14%, n = 3). The method was evaluated in terms of selectivity, sensitivity, matrix-effect, instrumental carry-over,
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