Oliva Alexis, Santoveña Ana, Fariña José, Llabrés Matías
Dpto. Ingeniería Química y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de La Laguna, 38200 La Laguna, Tenerife, Spain.
J Pharm Biomed Anal. 2003 Sep 19;33(2):145-55. doi: 10.1016/s0731-7085(03)00223-1.
Size-exclusion chromatography (SEC) was used to monitor the time-course of protein degradation induced by high shear rates during the formulation and manufacture of controlled-release pharmaceutical dosage forms. SEC with multi-angle laser light-scattering (MALLS) detection was used to characterize the aggregation products, determining their absolute molecular weight. A stability-indicating method was developed and validated to obtain reliable drug degradation data. The results obtained according to the ICH guidelines confirm that the system and methods proposed are suitable for their intended use. The degradation kinetics are influenced by the type of protein and the effect of the shear rate on their stability. Reversible pseudo-first order degradation kinetics were observed for bovine beta-lactoglobulin, whereas for human (HSA) and bovine serum albumin (BSA), a monomer-dimer transition was observed, independently of the rate of shear. However, trimer formation was also observed for HSA, especially at high shear rates. The kinetic model may thus be described as a two-step process: a monomer-dimer, and dimer-trimer transition.
尺寸排阻色谱法（SEC）用于监测控释药物剂型的配制和生产过程中高剪切速率诱导的蛋白质降解的时间进程。采用多角度激光散射（MALLS）检测的尺寸排阻色谱法对聚集产物进行表征，确定其绝对分子量。开发并验证了一种稳定性指示方法，以获得可靠的药物降解数据。根据国际人用药品注册技术协调会（ICH）指南获得的结果证实，所提出的系统和方法适用于其预期用途。降解动力学受蛋白质类型以及剪切速率对其稳定性的影响。观察到牛β-乳球蛋白呈现可逆的假一级降解动力学，而对于人血清白蛋白（HSA）和牛血清白蛋白（BSA），则观察到单体-二聚体转变，且与剪切速率无关。然而，在HSA中也观察到三聚体的形成，尤其是在高剪切速率下。因此，动力学模型可描述为一个两步过程：单体-二聚体以及二聚体-三聚体转变。
