Douglas Pharma US Inc, 1035 Louis Drive, Warminster, PA, USA 18974; Department of Chemistry, GITAM School of Science, GITAM deemed to be University, Hyderabad, Telangana, India 502329.
Department of Chemistry, GITAM School of Science, GITAM deemed to be University, Hyderabad, Telangana, India 502329.
J Chromatogr A. 2022 Aug 30;1679:463380. doi: 10.1016/j.chroma.2022.463380. Epub 2022 Jul 30.
An anti-inflammatory skin condition is treated with fluocinolone acetonide (FLA), a synthetic corticoid. The current study aims to develop a stability-indicating UPLC method for the determination of impurities present in fluocinolone acetonide and its topical oil formulation. The method development was performed by implementing Analytical Quality by Design (AQbD) and green chemistry principles. A detailed risk assessment was conducted based on the cause-and-effect relationship. d-optimal split-plot design was employed to screen the critical method parameters (CMPs). The central composite design (CCD) was employed to optimize the final method conditions. p-values for the model and lack of fit were <0.0001 and >0.05, respectively, which indicates the best fit statistical model for the studied responses (peak resolutions R1 - R5). The critical method attributes (CMAs) and CMPs such as the ratio of ACN: Water in mobile phase-B as 600:400 (v/v), the ratio of mobile phase-A & B in initial gradient program as 60:40, flow rate as 0.3 mL min, and column oven temperature as 50 °C were optimized from the CCD. The best possible separation among all components was achieved with a gradient elution using Waters Acquity UPLC HSS C, 100 mm × 2.1 mm, 1.8 µm analytical column. The optimized gradient program is time (min)/%B: 0.0/40, 1.5/40, 6.0/60, 8.0/70, 9.0/80, 12.0/100, 15.0/100, 15.1/40 & 18.0/40. Optimization of diluent is highly critical for any oil-based formulations. The experimental results show that acetonitrile is the most suitable diluent for the current study. The method validation was executed in compliance with ICH and USP 〈1225〉 guidelines. Mean recovery of the impurities ranged between 95.7 and 105.7%, the correlation coefficient(r) was> 0.999, the RSD values (n = 6) ranged between 0.9 - 3.2% across the range for LOQ - 150% levels. The peaks from the specificity study did not interfere with the known and active analyte peaks. The major degradation products were identified as Imp-C, B, and A, and established their degradation pathways from FLA based on the stress studies. The method greenness was evaluated using GAPI, AGREE and analytical eco scale and found that the method is green.
一种名为氟轻松丙酮（FLA）的合成皮质激素被用于治疗抗炎性皮肤病。本研究旨在建立一种稳定性指示的 UPLC 方法，用于测定氟轻松丙酮及其局部油制剂中的杂质。方法开发通过实施分析质量源于设计（AQbD）和绿色化学原则进行。基于因果关系进行了详细的风险评估。采用 d-最优分割-区组设计筛选关键方法参数（CMPs）。采用中心组合设计（CCD）优化最终方法条件。模型和缺乏拟合的 p 值分别小于 0.0001 和大于 0.05，表明该研究响应（峰分辨率 R1-R5）的最佳拟合统计模型。从 CCD 中优化了关键方法属性（CMAs）和 CMPs，如流动相-B 中 ACN：水的比例为 600:400（v/v）、初始梯度程序中流动相-A 和 B 的比例为 60:40、流速为 0.3 mL min 和柱温箱温度为 50°C。使用 Waters Acquity UPLC HSS C、100mm×2.1mm、1.8µm 分析柱进行梯度洗脱，实现了所有成分的最佳分离。优化的梯度程序是时间（min）/%B：0.0/40、1.5/40、6.0/60、8.0/70、9.0/80、12.0/100、15.0/100、15.1/40 和 18.0/40。对于任何油基制剂，稀释剂的优化都非常关键。实验结果表明，乙腈是本研究最适合的稀释剂。方法验证符合 ICH 和 USP 〈1225〉指南。杂质的平均回收率在 95.7%至 105.7%之间，相关系数（r）大于 0.999，在 LOQ-150%水平范围内，RSD 值（n=6）在 0.9-3.2%之间。专属性研究中的峰没有干扰已知和活性分析物的峰。确定了主要降解产物为 Imp-C、B 和 A，并根据应激研究确定了它们从 FLA 产生的降解途径。使用 GAPI、AGREE 和分析生态尺度评估方法的绿色度，发现该方法是绿色的。
