Abstract
A simple, selective, sensitive, accurate, and precise RP-HPLC method was developed for determination of Ethamsylate (ET) in human breast milk samples. Sample preparation consisted of mixing equal volumes of milk with acetonitrile followed by filtration through 3000 Da molecular mass cut-off filters. The method uses C18 column (5 µm particle size) at ambient temperature with mobile phase consisting of 7.0 mM dibasic sodium phosphate, adjusted to pH 3.0 using phosphoric acid 8.15 mM tetraheptylammonium bromide in acetonitrile (50:50 v/v), at a flow rate of 1.0 mL/min. Quantitation was achieved with UV detection at 300 nm based on peak area with linear calibration curve at the concentration range of 0.08–20 µg/mL. The proposed method was completely validated according to the FDA guidelines and was applied for the determination of ET in breast milk obtained from breast feeding mothers after oral administration of ET. The breast milk secretion pattern of ET was investigated for breastfeeding women using the developed method.
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