Roy Chinmoy, Chakrabarty Jitamanyu
Analytical Research and Development, Integrated Product Development, Dr. Reddy's Laboratories Ltd., Bachupally, Hyderabad-500090, Andhra Pradesh, India. ; Department of Chemistry, National Institute of Technology, Durgapur-713209, West Bengal, India.
Department of Chemistry, National Institute of Technology, Durgapur-713209, West Bengal, India.
Sci Pharm. 2014 Feb 27;82(3):519-39. doi: 10.3797/scipharm.1312-20. Print 2014 Jul-Sep.
A stability-indicating RP-HPLC method has been developed and validated for the simultaneous determination of methylparaben (MP), propylparaben (PP), diethylamino hydroxybenzoyl hexyl benzoate (DAHHB), and octinoxate (OCT) in topical pharmaceutical formulation. The desired chromatographic separation was achieved on the Kinetex(TM) C18 (250 × 4.6 mm, 5 μm) column using gradient elution at 257 nm detection wavelength. The optimized mobile phase consisted of a buffer : acetonitrile : tetrahydrofuran (60 : 30 : 10, v/v/v) as solvent A and acetonitrile : tetrahydrofuran (70 : 30, v/v) as solvent B. The method showed linearity over the range of 0.19-148.4 μg/mL, 0.23-15.3 μg/mL, 1.97-600.5 μg/mL, and 1.85-451.5 μg/mL for MP, PP, DAHHB, and OCT, respectively. Recovery for all the components was found to be in the range of 98-102%. The stability-indicating capability of the developed method was established by analysing the forced degradation samples in which the spectral purity of MP, PP, DAHHB, and OCT, along with the separation of the degradation products from the analyte peaks, was achieved. The proposed method was successfully applied for the quantitative determination of MP, PP, DAHHB, and OCT in the lotion sample. The design expert with ANOVA software with the linear model was applied and a 2(4) full factorial design was employed to estimate the model coefficients and also to check the robustness of the method. Results of the two-level full factorial design, 2(4) with 20 runs including four centrepoint analysis based on the variance analysis (ANOVA), demonstrated that all four factors, as well as the interactions of resolution between DAHHB and OCT are statistically significant.
已开发并验证了一种稳定性指示反相高效液相色谱法，用于同时测定局部用药物制剂中的对羟基苯甲酸甲酯（MP）、对羟基苯甲酸丙酯（PP）、二乙氨基羟基苯甲酰基己基苯甲酸酯（DAHHB）和桂皮酸盐（OCT）。在Kinetex(TM) C18（250×4.6 mm，5μm）色谱柱上，于257 nm检测波长下采用梯度洗脱实现了所需的色谱分离。优化后的流动相由缓冲液：乙腈：四氢呋喃（60:30:10，v/v/v）作为溶剂A和乙腈：四氢呋喃（70:30，v/v）作为溶剂B组成。该方法在0.19 - 148.4μg/mL、0.23 - 15.3μg/mL、1.97 - 600.5μg/mL和1.85 - 451.5μg/mL范围内分别显示出线性关系，对应MP、PP、DAHHB和OCT。所有成分的回收率在98% - 102%范围内。通过分析强制降解样品建立了所开发方法的稳定性指示能力，其中实现了MP、PP、DAHHB和OCT的光谱纯度以及降解产物与分析物峰的分离。所提出的方法成功应用于洗剂样品中MP、PP、DAHHB和OCT的定量测定。应用带有线性模型的设计专家ANOVA软件，并采用2(4)全因子设计来估计模型系数并检查方法的稳健性。基于方差分析（ANOVA）的包含四个中心点分析的20次运行的二级全因子设计2(4)结果表明，所有四个因素以及DAHHB和OCT之间的分辨率相互作用在统计学上均具有显著性。
