Method Development and Validation of Promethazine HCl by Using RP-HPLC and Its Forced Degradation Studies.

IF 1.8 4区医学 Q4 BIOCHEMICAL RESEARCH METHODS

Assay and drug development technologies Pub Date : 2026-09-22 DOI:10.1177/1540658X261486549

Deepshikha Nirwan, Kandasamy Nagarajan, Parul Grover, Snigdha Bhardwaj, Ashutosh Mishra

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Abstract

A simple, rapid, sensitive, and stability-indicating reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the quantitative determination of promethazine hydrochloride (HCl) in pharmaceutical formulations. Chromatographic separation was achieved using a SunFire® C18 column (4.6 × 250 mm, 5 µm) with a mobile phase consisting of acetonitrile and 0.1% orthophosphoric acid in water (80:20, v/v) at a flow rate of 1.0 mL/min. Detection was performed at 249 nm using a photodiode array detector. The developed method was validated according to the International Council for Harmonisation Q2 guidelines with respect to specificity, linearity, precision, accuracy, robustness, limit of detection, and limit of quantitation.The method exhibited good linearity over the concentration range of 2-64 µg/mL with a correlation coefficient (R²) of 0.9993. The percentage relative standard deviation for precision studies was below 2%, indicating satisfactory repeatability and reproducibility. Forced degradation studies were performed under acidic, alkaline, neutral, oxidative, and thermal stress conditions to evaluate the stability indicating capability of the method. Additional chromatographic peaks corresponding to degradation products were observed in acid, base, neutral, and oxidative stress conditions, while minimal degradation was observed under thermal stress. Furthermore, liquid chromatography-mass spectrometry (LC-MS) analysis was carried out to characterize the degradation products formed during stress studies. LC-MS results indicated that alkaline stress produced the highest degree of degradation, whereas thermal stress generated only minor residual impurities, confirming the relative stability of promethazine HCl under thermal conditions. The developed RP-HPLC method was found to be suitable for routine quality control analysis and stability assessment of promethazine HCl in pharmaceutical formulations.

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方法采用反相高效液相色谱法对盐酸异丙嗪进行开发、验证及强制降解研究。

建立了一种简便、快速、灵敏、稳定性好的反相高效液相色谱法（RP-HPLC），并对其进行了验证。色谱分离采用SunFire®C18色谱柱（4.6 × 250 mm, 5µm），流动相为乙腈和0.1%正磷酸水溶液（80:20,v/v），流速为1.0 mL/min。使用光电二极管阵列检测器在249 nm处进行检测。根据国际协调委员会Q2指南对所开发的方法进行了特异性、线性、精密度、准确性、鲁棒性、检出限和定量限的验证。在2 ~ 64µg/mL范围内线性良好，相关系数（R2）为0.9993。精密度研究的相对标准偏差小于2%，重复性和再现性令人满意。在酸性、碱性、中性、氧化和热应力条件下进行了强制降解研究，以评估该方法的稳定性和性能。在酸、碱、中性和氧化应激条件下观察到降解产物对应的附加色谱峰，而在热应激条件下观察到最小的降解。此外，采用液相色谱-质谱（LC-MS）分析对应力研究过程中形成的降解产物进行了表征。LC-MS结果表明，碱胁迫降解程度最高，而热胁迫仅产生少量残留杂质，证实了异丙嗪HCl在热条件下的相对稳定性。所建立的反相高效液相色谱法适用于盐酸异丙嗪制剂的常规质量控制分析和稳定性评价。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Assay and drug development technologies 医学-生化研究方法

CiteScore

3.60

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： ASSAY and Drug Development Technologies provides access to novel techniques and robust tools that enable critical advances in early-stage screening. This research published in the Journal leads to important therapeutics and platforms for drug discovery and development. This reputable peer-reviewed journal features original papers application-oriented technology reviews, topical issues on novel and burgeoning areas of research, and reports in methodology and technology application. ASSAY and Drug Development Technologies coverage includes: -Assay design, target development, and high-throughput technologies- Hit to Lead optimization and medicinal chemistry through preclinical candidate selection- Lab automation, sample management, bioinformatics, data mining, virtual screening, and data analysis- Approaches to assays configured for gene families, inherited, and infectious diseases- Assays and strategies for adapting model organisms to drug discovery- The use of stem cells as models of disease- Translation of phenotypic outputs to target identification- Exploration and mechanistic studies of the technical basis for assay and screening artifacts