Ram Vijay R, Dave Pragnesh N, Joshi Hitendra S
Department of Chemistry, Krantiguru Shyamji Krishna Verma Kachchh University, Bhuj-370 001, Gujarat, India.
J Chromatogr Sci. 2012 Sep;50(8):721-6. doi: 10.1093/chromsci/bms062. Epub 2012 May 28.
The objective of the current study was to develop and validate a simple, precise and accurate isocratic stability-indicating reversed-phase high-performance liquid chromatography (RP-HPLC) assay method for the determination of spironolactone and furosemide in solid pharmaceutical dosage forms. Isocratic RP-HPLC separation was achieved on an SGE 150 × 4.6 mm SS Wakosil II 5C8RS 5-μm column using a mobile phase of acetonitrile-ammonium acetate buffer (50:50, v/v) at a flow rate of 1.0 mL/min. The detection was carried out at 254 nm using a photodiode array detector. The drug was subject to oxidation, hydrolysis, photolysis and heat to apply stress conditions. The method was validated for specificity, linearity, precision, accuracy, robustness and solution stability. The method was found to be linear in the drug concentration range of 40-160 µg/mL with correlation coefficients of 0.9977 and 0.9953 for spironolactone and furosemide, respectively. The precision (relative standard deviation; RSD) among a six-sample preparation was 0.87% and 1.1% for spironolactone and furosemide, respectively. Repeatability and intermediate precision (RSD) among a six-sample preparation were 0.46% and 0.20% for spironolactone and furosemide, respectively. The accuracy (recovery) was between 98.05 and 100.17% and 99.07 and 100.58% for spironolactone and furosemide, respectively. Degradation products produced as a result of stress studies did not interfere with the detection of spironolactone and furosemide; therefore, the assay can be considered to be stability-indicating.
本研究的目的是开发并验证一种简单、精确且准确的等度稳定性指示反相高效液相色谱（RP-HPLC）测定方法，用于测定固体药物剂型中的螺内酯和呋塞米。在SGE 150×4.6 mm SS Wakosil II 5C8RS 5-μm色谱柱上进行等度RP-HPLC分离，流动相为乙腈-醋酸铵缓冲液（50:50，v/v），流速为1.0 mL/min。使用光电二极管阵列检测器在254 nm波长处进行检测。对药物进行氧化、水解、光解和加热处理以施加强制降解条件。该方法针对专属性、线性、精密度、准确度、稳健性和溶液稳定性进行了验证。结果发现该方法在40 - 160 µg/mL的药物浓度范围内呈线性，螺内酯和呋塞米的相关系数分别为0.9977和0.9953。六份样品制备的精密度（相对标准偏差；RSD），螺内酯和呋塞米分别为0.87%和1.1%。六份样品制备的重复性和中间精密度（RSD），螺内酯和呋塞米分别为0.46%和0.20%。准确度（回收率），螺内酯在98.05%至100.17%之间，呋塞米在99.07%至100.58%之间。强制降解研究产生的降解产物不干扰螺内酯和呋塞米的检测；因此，该测定法可被认为是稳定性指示的。
