Stability-indicating high performance liquid chromatography (HPLC), thin-layer chromatography (TLC) and first-derivative of ratio spectra (1DD) methods are developed for the determination of piretanide in presence of its alkaline induced degradates. HPLC method depends on separation of piretanide from its degradates on mu-Bondapak C18 column using methanol:water:acetic acid (70:30:1, v/v/v) as a mobile phase at flow rate 1.0 ml/min and UV detector at 275 nm. TLC densitometic method is based on the difference in Rf-values between the intact drug and its degradates on thin-layer silica gel. Iso-propanol:ammonia 33% (8:2, v/v) was used as a developing mobile phase and the chromatogram was scanned at 275 nm. The derivative of ratio spectra method (1DD) depends on the measurement of the absorbance at 288 nm in the first-derivative of ratio spectra for the determination of the cited drug in the presence of its degradates. Calibration graphs of the three suggested methods are linear in the concentration ranges 0.02-0.3 microg/20 microl, 0.5-10 microg/spot and 5-50 microg/ml, with mean percentage recovery 99.27+/-0.52, 99,17+/-1.01 and 99.65+/-1.01%, respectively. The three proposed methods were successfully applied for the determination of piretanide in bulk powder, laboratory-prepared mixtures and pharmaceutical dosage form with good accuracy and precision. The results were statistically analyzed and compared with those obtained by the official method. Validation of the method was determined with favourable specificity, linearity, precision, and accuracy was assessed by applying the standard addition technique.

中文翻译：

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在碱性诱导的降解物存在下测定吡咯他尼的稳定性指示方法。

建立了指示稳定性的高效液相色谱（HPLC），薄层色谱（TLC）和比率谱一阶导数（1DD）方法，用于测定吡咯他尼在碱诱导的降解物中的存在。HPLC方法取决于使用甲醇：水：乙酸（70：30：1，v / v / v）作为流动相，流速为1.0 ml / min和UV检测器在mu-Bondapak C18色谱柱上分离吡咯他尼类的产物在275 nm处。TLC光密度法基于完整药物及其在薄层硅胶上降解的药物之间的Rf值差异。使用33％的异丙醇：氨水（8：2，v / v）作为展开流动相，并在275 nm处扫描色谱图。比率光谱的导数方法（1DD）取决于比率光谱的一阶导数在288 nm处的吸光度的测定，用于在存在降解物的情况下确定所引用的药物。三种建议方法的校正曲线在0.02-0.3 microg / 20 microl，0.5-10 microg / spot和5-50 microg / ml的浓度范围内呈线性关系，平均回收率99.27 +/- 0.52,99,17 + / -1.01和99.65 +/- 1.01％。所提出的三种方法已成功地用于散装粉末，实验室制备的混合物和药物剂型中吡咯他尼的测定，具有良好的准确度和精密度。对结果进行统计分析，并与通过官方方法获得的结果进行比较。该方法的验证具有良好的特异性，线性，精密度，