Flórez Borges Paloma, Pérez Lozano Pilar, García Montoya Encarna, Miñarro Montserrat, Ticó Josep R, Jo Enric, Suñe Negre Josep M
Pharmacy and Pharmaceutical Technology Department, Faculty of Pharmacy, University of Barcelona, Avda Joan XXIII s/n 08028, Barcelona, Spain.
Reig Jofre Group, c. Gran Capitá 6 08970, Sant Joan Despi, Barcelona, Spain.
Daru. 2014 Dec 9;22(1):82. doi: 10.1186/s40199-014-0082-5.
A new, simple and accurate stability-indicating reverse phase high performance liquid chromatography method was developed and validated during the early stage of drug development of an oral lyophilizate dosage form of cetirizine dihydrochloride.
For RP-HPLC analysis it was used an Eclipse XDB C8 column 150 mm × 4.6 mm, 5 μm (Agilent columns, Barcelona, Spain) as the stationary phase with a mobile phase consisted of a mixture of 0.2 M K2HPO4 pH 7.00 and acetonitrile (65:35, v/v) at a flow rate of 1 mL min (-1). Detection was performed at 230 nm using diode array detector. The method was validated in accordance with ICH guidelines with respect to linearity, accuracy, precision, specificity, limit of detection and quantification.
The method results in excellent separation between the drug substance and its stress-induced degradation products. The peak purity factor is >950 for the drug substance after all types of stress, which confirms the complete separation of the drug substance peak from its stress induced degradation products. Regression analysis showed r(2) > 0.999 for cetirizine dihydrochloride in the concentration range of 650 μg mL (-1) to 350 μg mL(-1) for drug substance assay and a r(2) > 0.999 in the concentration range of 0.25 μg mL(-1) to 5 μg mL(-1) for degradation products. The method presents a limit of detection of 0.056 μg mL (-1) and a limit of quantification of 0.25 μg mL(-1). The obtained results for precision and accuracy for drug substance and degradation products are within the specifications established for the validation of the method.
The proposed stability-indicating method developed in the early phase of drug development proved to be a simple, sensitive, accurate, precise, reproducible and therefore useful for the following stages of the cetirizine dihydrochloride oral lyophilizate dosage form development.
在盐酸西替利嗪口服冻干剂型药物研发的早期阶段，开发并验证了一种新的、简单且准确的稳定性指示反相高效液相色谱法。
对于反相高效液相色谱分析，使用Eclipse XDB C8柱（150 mm×4.6 mm，5μm，安捷伦色谱柱，西班牙巴塞罗那）作为固定相，流动相由0.2 M磷酸氢二钾（pH 7.00）和乙腈（65:35，v/v）的混合物组成，流速为1 mL·min⁻¹。使用二极管阵列检测器在230 nm处进行检测。该方法根据国际协调会议（ICH）指南在线性、准确度、精密度、特异性、检测限和定量限方面进行了验证。
该方法实现了原料药与其应激诱导降解产物之间的良好分离。在所有类型的应激后，原料药的峰纯度因子>950，这证实了原料药峰与其应激诱导降解产物的完全分离。回归分析表明，对于原料药含量测定，盐酸西替利嗪在650μg·mL⁻¹至350μg·mL⁻¹浓度范围内的r²>0.999，对于降解产物在0.25μg·mL⁻¹至5μg·mL⁻¹浓度范围内的r²>0.999。该方法的检测限为0.056μg·mL⁻¹，定量限为0.25μg·mL⁻¹。原料药和降解产物的精密度和准确度所得结果在该方法验证所规定的规格范围内。
在药物研发早期阶段开发的所提出的稳定性指示方法被证明是简单、灵敏、准确、精密、可重现的，因此对于盐酸西替利嗪口服冻干剂型研发的后续阶段是有用的。
