Dr. Risch Ostschweiz AG, Buchs, Switzerland.
Roche Diagnostics GmbH, Penzberg, Germany.
Clin Chem Lab Med. 2024 Mar 29;62(7):1327-1338. doi: 10.1515/cclm-2023-1032. Print 2024 Jun 25.
Primidone is an anticonvulsive drug used in the treatment of epilepsy and essential tremor. It offers beneficial effects in controlling seizures, but its usage is also associated with possible side effects. To ensure optimal therapy, it is crucial to measure its concentration through accurate quantification methods. Therefore, our main goal was to develop and validate a new reference measurement procedure (RMP) for accurately measuring primidone levels in human serum and plasma.
In our study, we focused on the separation of primidone from both known and unknown interferences using a C18 column. To achieve accurate sample preparation, we developed a protocol involving protein precipitation followed by a high dilution step. The validation of the assay and determination of measurement uncertainty were carried out following guidelines from organizations such as the Clinical and Laboratory Standards Institute, the International Conference on Harmonization, and the Guide to the Expression of Uncertainty in Measurement. These rigorous validation processes ensure the reliability and accuracy of our method for quantifying primidone levels in human serum and plasma samples.
The RMP was shown to be highly selective and specific, with no evidence of matrix interference. It can be used to quantify primidone in the range of 0.150-30.0 μg/mL. Intermediate precision was less than 4.0 %, and repeatability CV ranged from 1.0 to 3.3 % across all concentration levels. The relative mean bias ranged from 0.1 to 3.9 % for native serum levels, and from -2.6 to 2.8 % for lithium-heparin plasma levels. The measurement uncertainties for single measurements and target value assignment were 1.5-4.1 % and 0.9-1.0 %, respectively.
In this study, we introduce an innovative LC-MS/MS-based candidate RMP specifically designed for primidone in human serum and plasma. Our RMP offers a traceable platform, facilitating the standardization of routine assays and enabling the evaluation of clinically relevant samples. With this novel approach, we aim to enhance the accuracy and reliability of primidone measurements, ultimately benefiting the field of clinical research and patient care.
扑米酮是一种抗惊厥药物，用于治疗癫痫和特发性震颤。它在控制癫痫发作方面具有有益的效果，但它的使用也与可能的副作用有关。为了确保最佳的治疗效果，通过准确的定量方法来测量其浓度是至关重要的。因此，我们的主要目标是开发和验证一种新的参考测量程序（RMP），以准确测量人血清和血浆中的扑米酮水平。
在我们的研究中，我们专注于使用 C18 柱从已知和未知干扰物中分离扑米酮。为了实现准确的样品制备，我们开发了一种涉及蛋白质沉淀和高稀释步骤的方案。该测定的验证和测量不确定度的确定是按照临床和实验室标准协会、国际协调会议和测量不确定度表示指南等组织的指南进行的。这些严格的验证过程确保了我们用于定量人血清和血浆样品中扑米酮水平的方法的可靠性和准确性。
该 RMP 被证明具有高度的选择性和特异性，没有基质干扰的证据。它可以用于定量 0.150-30.0μg/mL 范围内的扑米酮。中间精密度小于 4.0%，所有浓度水平的重复性 CV 范围为 1.0-3.3%。内源性血清水平的相对平均偏差范围为 0.1-3.9%，锂-肝素血浆水平的相对平均偏差范围为-2.6-2.8%。单次测量和目标值赋值的测量不确定度分别为 1.5-4.1%和 0.9-1.0%。
在这项研究中，我们引入了一种基于 LC-MS/MS 的创新候选 RMP，专门用于人血清和血浆中的扑米酮。我们的 RMP 提供了一个可溯源的平台，促进了常规测定的标准化，并能够评估临床相关样本。通过这种新方法，我们旨在提高扑米酮测量的准确性和可靠性，最终造福临床研究和患者护理领域。
