Gupta Sakshi, Bansal Gulshan
Punjabi University, Department of Pharmaceutical Sciences and Drug Research, Patiala 147002, Punjab, India.
J AOAC Int. 2011 Mar-Apr;94(2):523-30.
A selective stability-indicating HPLC-UV method for simultaneous determination of glipizide and four impurities (DPs I-IV) formed under hydrolytic conditions was developed and validated. The drug and impurities were resolved on an XTerra C18 column (250 x 4.5 mm id) in a single gradient run using buffer (0.005 M KH2PO4; pH 3.0)-methanol (60 + 40, v/v; mobile phase A) and (20 + 80, v/v; mobile phase B) at a flow rate of 0.5 mL/min with 230 nm detection wavelength. The method was linear across concentration ranges of 0.2-100, 0.1-100, 0.5-100, 0.2-100, and 0.1-50 microg/mL for glipizide and DPs I-IV, respectively. The RSD for intraday and interday precision for the drug and impurities was < 1 and < 1.2%, respectively. Satisfactory recoveries (96.58-99.97%) of each of the three concentrations selected across the linearity range of each analyte were obtained, proving the method was sufficiently accurate. The LOD was 0.07, 0.05, 0.16, 0.08, and 0.05 microg/mL and the LOQ was 0.20, 0.14, 0.50, 0.23, and 0.14 microg/mL for the drug and DPs I-IV, respectively. Each peak was resolved with resolution of > 2 from the nearest peak. Insignificant changes in retention time (< 4%) and calculated amount (< 1.65%) of drug and each impurity upon small but deliberate changes in various chromatographic parameters were observed, suggesting the method was robust. The method was applied successfully to stability testing of glipizide tablets.
建立并验证了一种选择性的稳定性指示高效液相色谱 - 紫外检测法，用于同时测定格列吡嗪及在水解条件下形成的四种杂质（降解产物I - IV）。药物和杂质在XTerra C18柱（250×4.5 mm内径）上，通过单一梯度洗脱进行分离，流动相A为缓冲液（0.005 M KH2PO4；pH 3.0） - 甲醇（60 + 40，v/v），流动相B为缓冲液（0.005 M KH2PO4；pH 3.0） - 甲醇（20 + 80，v/v），流速为0.5 mL/min，检测波长为230 nm。该方法在格列吡嗪及降解产物I - IV的浓度范围分别为0.2 - 100、0.1 - 100、0.5 - 100、0.2 - 100和0.1 - 50 μg/mL时呈线性。药物和杂质的日内及日间精密度相对标准偏差分别<1%和<1.2%。在每个分析物线性范围内选择的三个浓度下，各物质均获得了满意的回收率（96.58 - 99.97%），证明该方法足够准确。药物及降解产物I - IV的检测限分别为0.07、0.05、0.16、0.08和0.05 μg/mL，定量限分别为0.20、0.14、0.50、0.23和0.14 μg/mL。每个峰与最近峰的分离度>2。在各种色谱参数进行微小但有意的改变后，观察到药物和各杂质的保留时间（<4%）和计算量（<1.65%）变化不显著，表明该方法具有稳健性。该方法成功应用于格列吡嗪片的稳定性测试。
