Abstract：
β-propiolactone (BPL) is an alkylating agent used for inactivation of biological samples such as vaccines. Due to its known carcinogenic properties, complete hydrolysis of BPL is essential, and the detection of trace amounts is crucial. In this study a novel High-Performance Liquid Chromatography-Ultraviolet (HPLC-UV) method was developed. Rhodamine B hydrazide (RBH) was synthesized and utilized as a derivatizing reagent to react with BPL. The reaction was optimized in a weak acidic solution, resulting in a high yield. The separation of the RBH-derivatized BPL was achieved on a C8 column and detected by a UV detector at a wavelength of 560 nm. The method\'s validation demonstrated a high linearity (r2 > 0.99) over a concentration range of 0.5-50 µg/mL, with detection and quantification limits of 0.17 µg/mL and 0.5 µg/mL, respectively. The average recovery of samples was 85.20 % with a relative standard deviation (RSD) of 1.75 %. This method was successfully applied for BPL residue analysis in inactivated COVID-19 vaccines. This novel derivatization method offers a promising solution for monitoring BPL residues in the vaccine production process for quality control purposes and compliance with regulatory standards.
摘要：
β-丙内酯(BPL)是用于灭活生物样品如疫苗的烷化剂。由于其已知的致癌特性，BPL的完全水解是必不可少的，痕量的检测至关重要。在这项研究中，开发了一种新型的高效液相色谱-紫外线（HPLC-UV）方法。合成了罗丹明B酰肼（RBH），并将其用作衍生试剂与BPL反应。在弱酸性溶液中优化反应，导致高产量。RBH衍生的BPL的分离在C8柱上实现，并通过波长为560nm的UV检测器进行检测。该方法的验证表明在0.5-50µg/mL的浓度范围内具有高线性（r2>0.99），检测和定量限为0.17µg/mL和0.5µg/mL，分别。样品的平均回收率为85.20%,相对标准偏差(RSD)为1.75%。该方法已成功应用于COVID-19灭活疫苗中BPL的残留分析。这种新颖的衍生方法为监测疫苗生产过程中的BPL残留提供了一种有前途的解决方案，用于质量控制目的和符合监管标准。
