Panjab University, UGC Centre of Advanced Studies, University Institute of Pharmaceutical Sciences, Chandigarh, 160 014, India.
Parul University, Parul Institute of Pharmacy, Vadodara, Gujarat, 391760, India.
J AOAC Int. 2021 Jun 12;104(3):620-632. doi: 10.1093/jaoacint/qsab014.
Ceftazidime, a third-generation cephalosporin, is widely used in the treatment of lung infections, often given as "off-label" nebulization. There is a need to develop a sensitive and robust analytical method to compute aerodynamic properties of ceftazidime following nebulization.
The current study entails development of a simple, accurate, and sensitive HPLC method for ceftazidime estimation, employing the principles of analytical quality-by-design (AQbD) and Monte Carlo simulations.
Selection of critical material attributes (CMAs) affecting method performance was accomplished by factor screening exercises. Subsequently, the influential CMAs, i.e., mobile phase ratio and flow rate, were systemically optimized using a face-centered cubic design for the chosen critical analytical attributes (CAAs). The factor relationship(s) between CMAs and CAAs was explored employing a 3 D-response surface and 2 D-contour plots, followed by numerical as well as graphical optimization, for establishing the optimal chromatographic conditions. The obtained method operable design region was validated by Monte Carlo simulations for defect rate analysis.
The optimized HPLC conditions for estimating ceftazidime were acetonitrile to acetic acid solution (75:25) as mobile phase at a flow rate of 0.7 mL/min, leading to Rt of 4.5 min and peak tailing ≤2. Validation studies, as per International Conference on Harmonization Q2(R1) guidance, demonstrated high sensitivity, accuracy, and efficiency of the developed analytical method with an LOD of 0.075 and LOQ of 0.227 µg/mL. Application of this chromatographic method was extrapolated for determining aerodynamic performance by nebulizing ceftazidime at a flow rate of 15 L/min using a next-generation impactor. The study indicated superior performance, sensitivity, and specificity of the developed analytical system for quantifying ceftazidime.
Application of an AQbD approach, coupled with Monte Carlo simulations, aided in developing a robust HPLC method for estimationof ceftazidime per se and on various stages of impactor.
(i) QbD-enabled development of robust RP-HPLC method for ceftazidime quantification, (ii) Analytical method optimization employing Risk Assessment and Design of Experiments, (iii) Design space verification and defect rate analysis using Monte Carlo simulations, (iv) Chromatographic method validation as per ICH Q2 R1 guidelines and (v) Quantitative estimation of ceftazidime on various stages of impactor.
头孢他啶是一种第三代头孢菌素，广泛用于治疗肺部感染，常作为“超适应证”雾化剂使用。需要开发一种灵敏且稳健的分析方法来计算雾化后头孢他啶的空气动力学特性。
本研究旨在开发一种简单、准确、灵敏的 HPLC 法用于头孢他啶的估算，采用分析质量源于设计（AQbD）和蒙特卡罗模拟的原理。
通过因子筛选实验选择影响方法性能的关键物料属性（CMA）。随后，系统地优化了具有影响力的 CMA，即流动相比例和流速，采用中心复合设计（CCD）针对所选关键分析属性（CAA）进行优化。使用 3D 响应面和 2D 等高线图探讨了 CMA 与 CAA 之间的关系，然后进行数值和图形优化，以确定最佳的色谱条件。通过蒙特卡罗模拟对获得的方法操作设计区域进行验证，进行缺陷率分析。
用于估算头孢他啶的优化 HPLC 条件为乙腈-乙酸溶液（75：25）作为流动相，流速为 0.7 mL/min，Rt 为 4.5 min，峰拖尾≤2。根据国际人用药品注册技术协调会（ICH）Q2（R1）指南进行的验证研究表明，所开发的分析方法具有高灵敏度、准确性和效率，检测限（LOD）为 0.075，定量限（LOQ）为 0.227 μg/mL。该色谱方法的应用可推断出通过以 15 L/min 的流速使用下一代撞击器雾化头孢他啶来确定空气动力学性能。该研究表明，所开发的分析系统在定量测定头孢他啶方面具有优越的性能、灵敏度和特异性。
应用 AQbD 方法并结合蒙特卡罗模拟，有助于开发一种用于自身和撞击器各个阶段估算头孢他啶的稳健 HPLC 方法。
（i）通过 QbD 实现稳健的 RP-HPLC 方法开发，用于头孢他啶的定量测定，（ii）使用风险评估和实验设计优化分析方法，（iii）通过蒙特卡罗模拟验证设计空间和缺陷率分析，（iv）根据 ICH Q2 R1 指南进行色谱方法验证，（v）在撞击器的各个阶段对头孢他啶进行定量估计。
