Faculty of Pharmacy, Department of Pharmaceutical Analytical Chemistry, University of Alexandria, El-Messalah, Alexandria 21521, Egypt.
J Pharm Biomed Anal. 2012 Mar 5;61:78-85. doi: 10.1016/j.jpba.2011.11.032. Epub 2011 Dec 6.
An efficient chromatographic method for the simultaneous determination of triamterene (TRI) and xipamide (XIP) in urine samples, based on high performance liquid chromatography with photodiode array detector (HPLC-DAD) has been developed. The HPLC separation was performed on a RP stainless-steel C-18 analytical column (250 mm × 4.6 mm, 5 μm) with a gradient elution system of 0.05 M phosphate buffer adjusted to pH 4.0 and methanol as the mobile phase. The method was used to determine the urinary excretion profile and to calculate different urinary pharmacokinetic parameters following oral dose of their combination compared with single oral doses of each drug and hence comparing their bioavailability. Quantitation was performed using chlorthalidone as internal standard. The calibration graphs of each drug were rectilinear in the range of 0.2-40 μg/mL urine for TRI and 0.2-15 μg/mL urine for XIP. An HPLC-DAD method was also successfully developed for the simultaneous determination of the investigated drugs in pharmaceutical preparations. The methods were validated in terms of linearity, accuracy, precision, selectivity, limits of detection and quantitation and other aspects of analytical validation.
建立了一种高效液相色谱-光电二极管阵列检测（HPLC-DAD）法，用于同时测定尿液样品中的三甲氧苯嗪（TRI）和西米帕米（XIP）。HPLC 分离在 RP 不锈钢 C-18 分析柱（250mm×4.6mm，5μm）上进行，采用 0.05M 磷酸盐缓冲液（pH4.0）和甲醇作为流动相的梯度洗脱系统。该方法用于测定口服联合用药和单药后尿液排泄曲线，并计算不同的尿液药代动力学参数，从而比较它们的生物利用度。定量采用氯噻酮作为内标。每个药物的校准曲线在 0.2-40μg/mL 尿液范围内为 TRI，在 0.2-15μg/mL 尿液范围内为 XIP，均为线性。还成功建立了同时测定药物制剂中研究药物的 HPLC-DAD 法。该方法在线性、准确性、精密度、选择性、检测限和定量限以及分析验证的其他方面进行了验证。
