Hadad Ghada M, Abdel Salam Randa A, Emara Samy
Pharmaceutical Analytical Chemistry Department, Faculty of Pharmacy, University of Suez Canal, Ismailia 41522, Egypt.
J Chromatogr Sci. 2012 Jul;50(6):509-15. doi: 10.1093/chromsci/bms041. Epub 2012 Apr 23.
A high-performance liquid chromatographic method was optimized and validated for the determination of desacetyl nitazoxanide (tizoxanide), the main active metabolite of nitazoxanide in human plasma, urine and breast milk. The proposed method used a CN column with mobile phase consisting of acetonitrile-12mM ammonium acetate-diethylamine in the ratio of 30:70:0.1 (v/v/v) and buffered at pH 4.0 with acetic acid, with a flow rate of 1.5 mL/min. Quantitation was achieved with UV detection at 260 nm using nifuroxazide as internal standard. A simplified direct injection of urine samples without extraction in addition to the urinary excretion pattern were calculated using the proposed method. Also, the effectiveness of protein precipitation and a clean-up procedure were investigated for biological plasma and human breast milk samples. The validation study of the proposed method was successfully carried out in an assay range between 0.2 and 20 µg/mL.
优化并验证了一种高效液相色谱法，用于测定硝唑尼特的主要活性代谢物去乙酰硝唑尼特（替唑尼特）在人血浆、尿液和母乳中的含量。该方法采用氰基柱，流动相由乙腈 - 12mM醋酸铵 - 二乙胺按30:70:0.1（v/v/v）的比例组成，并用乙酸调节pH值至4.0，流速为1.5 mL/min。以硝呋柳胺作为内标，在260 nm处进行紫外检测实现定量分析。使用该方法计算了尿液样品简化的直接进样（无需萃取）以及尿排泄模式。此外，还研究了蛋白质沉淀和净化程序对生物血浆和人母乳样品的有效性。所提出方法的验证研究在0.2至20 µg/mL的测定范围内成功进行。
