高效液相色谱测定人血浆中米托坦浓度OA

Objective To establish a more clinically applicable high performance liquid chromatography(HPLC)method for the determination of mitotane concentration in human plasma,and to complete a method validation in accordance with the requirements of The International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use(ICH)《BIO ANALYTICAL METHOD VALIDATION AND STUDY SAMPLE ANALYSIS,M10》.Methods Using acetonitrile as the protein precipitant,plasma samples were pre-treated by protein precipitation and then analysed by HPLC using a Shim-pak CLC-ODS column(150.0 mm × 60.0 mm,5.0 μm),with a mobile phase of water-acetonitrile=20∶80(v/v,pH adjusted to 3.5 with phosphoric acid),a flow rate of 1.00 mL·min-1,a column temperature of 35 ℃,a UV detection wavelength of 230 nm,an injection volume of 50 μL,and a run time of 30 min.Results The standard curve of mitotane was y=1.08 × 10-1x-0.18 × 10-1(r=0.999 2),the linear range was 1.00-100.00 mg·L-1.The intra-batch accuracy was-8.80％-14.39％,with the precision was 0.07％-6.30％,the inter-batch accuracy was-1.63％-4.47％,with the precision was 6.67％-9.20％.The average extraction recovery rate was greater than 87.84％,and the plasma samples demonstrated good stability under various conditions with no significant carry-over.The commonly concomitant medications in clinical practice had no interference,and the whole blood remained stable when stored at room temperature for 2 h.Conclusion The method complied with the ICH M10 for method validation.It was highly sensitive,accurate,safe and simple to operate,and was suitable for the routine clinical TDM of mitotane concentrationin in human plasm.