Abucayon Erwin G, Sweeney Scott, Matyas Gary R
U.S. Military HIV Research Program, Center for Infectious Diseases Research, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, Maryland 20910, United States.
Henry M. Jackson Foundation for the Advancement of Military Medicine, 6720A Rockledge Drive, Bethesda, Maryland 20817, United States.
ACS Omega. 2024 Apr 17;9(17):19637-19644. doi: 10.1021/acsomega.4c01524. eCollection 2024 Apr 30.
Cholesterol, as one of the major components of liposomes, plays a critical role in modulating membrane bilayer permeability, fluidity, and structural stability. Controlling these quality attributes is essential to maintaining the efficacy and fitness of the liposomes in various applications. However, during the manufacture and storage of liposomes, cholesterol has a propensity to undergo oxidative degradation. Hence, an analytical tool that is capable of determining not only the identity and quantity of cholesterol but also its associated degradants is a prerequisite to effective process control and product quality and safety assessments. In this view, a new liquid chromatography electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method with parallel reaction monitoring (PRM) was developed and qualified to accurately quantify cholesterol and monitor the formation of 25-hydroxycholesterol degradant in liposomal drug formulations without the use of an isotopic internal standard (IS). The method was qualified according to the FDA Quality Guidance for Industry: Q2(R1). Study results showed that the method presents good specificity for cholesterol and 25-hydroxycholesterol detection in the liposomal matrix, good sensitivity characterized by LOD/LOQ in the nanomolar range, and accuracy within the range of 80 to 120%. The described method enables accurate evaluation of in-process and product release samples of Army Liposome Formulation with QS21 (ALFQ).
胆固醇作为脂质体的主要成分之一，在调节膜双层的通透性、流动性和结构稳定性方面起着关键作用。控制这些质量属性对于维持脂质体在各种应用中的功效和适用性至关重要。然而，在脂质体的制造和储存过程中，胆固醇容易发生氧化降解。因此，一种不仅能够测定胆固醇的身份和数量，还能测定其相关降解产物的分析工具，是有效过程控制以及产品质量和安全性评估的先决条件。鉴于此，开发并验证了一种采用平行反应监测（PRM）的新型液相色谱-电喷雾电离-串联质谱（LC-ESI-MS/MS）方法，该方法无需使用同位素内标（IS）就能准确量化脂质体药物制剂中的胆固醇，并监测25-羟基胆固醇降解产物的形成。该方法根据美国食品药品监督管理局（FDA）的《工业质量指南：Q2(R1)》进行了验证。研究结果表明，该方法在脂质体基质中对胆固醇和25-羟基胆固醇的检测具有良好的特异性，具有纳摩尔范围内的检测限/定量限所表征的良好灵敏度，以及80%至120%范围内的准确度。所述方法能够准确评估含QS21的陆军脂质体制剂（ALFQ）的过程中和产品放行样品。
