College of Pharmacy, Dankook University, Cheonan, Chungnam, South Korea.
Department of Pharmaceutical Engineering, Dankook University, Cheonan, Chungnam, South Korea.
PLoS One. 2024 Sep 6;19(9):e0309802. doi: 10.1371/journal.pone.0309802. eCollection 2024.
Donepezil (DPZ), a piperidine-based reversible cholinesterase inhibitor, finds extensive use in treating Alzheimer's disease (AD). Originally designed as an oral formulation, DPZ encounters drawbacks such as a brief duration of action and reduced treatment effectiveness in elderly patients with memory impairment or difficulty swallowing medications. To address these issues and improve patient compliance, researchers are actively exploring alternative DPZ formulations. Consequently, reliable methods are necessary to quantitate DPZ in biological samples for in vivo assessment. Therefore, we propose an efficient, sensitive, wide-dynamic, and cost-effective method for quantitating DPZ in rat plasma. Our method employs liquid-liquid extraction (LLE) followed by liquid chromatography and tandem mass spectrometry, enabling in vivo evaluation of novel DPZ formulations. Notably, our method requires only 20 μL of rat plasma and employs icopezil as the internal standard-a cost-effective compound with chemical similarity to DPZ. We meticulously optimized LLE conditions, taking into account factor interactions through design of experiments (DOE). Our rapid and straightforward extraction and purification involved using 500 μL of pure methyl tert-butyl ether to extract DPZ from the sample within five minutes. The dynamic range of the method extends from 0.5 ng/mL to 1,000 ng/mL, demonstrating excellent sensitivity and suitability for pharmacokinetic studies across diverse DPZ formulations. Following the FDA guidelines, we rigorously validated the developed method, evaluating selectivity, linearity (with a coefficient of determination ≥0.9999), accuracy (ranging from 96.0% to 109.6%), precision (≤13.9%), matrix effect (92.2% to 103.8%), recovery (98.5% to 106.8%), the lower limit of quantitation (0.5 ng/mL), and stability. Finally, we effectively employed the validated method for the long-term pharmacokinetic assessment of a DPZ formulation. We expect that this approach will make a substantial contribution to the advancement of new DPZ formulations, ultimately benefiting individuals afflicted by AD.
多奈哌齐（DPZ）是一种基于哌啶的可逆胆碱酯酶抑制剂，广泛用于治疗阿尔茨海默病（AD）。最初设计为口服制剂，DPZ 在记忆力减退或吞咽药物有困难的老年患者中，存在作用持续时间短和治疗效果降低等问题。为了解决这些问题并提高患者的依从性，研究人员正在积极探索 DPZ 的替代制剂。因此，需要可靠的方法来定量检测生物样本中的 DPZ，以便进行体内评估。因此，我们提出了一种高效、灵敏、宽动态范围和具有成本效益的方法，用于定量检测大鼠血浆中的 DPZ。我们的方法采用液-液萃取（LLE），然后进行液相色谱和串联质谱分析，能够对新型 DPZ 制剂进行体内评估。值得注意的是，我们的方法仅需要 20 μL 的大鼠血浆，并使用伊克培唑作为内标 - 一种与 DPZ 具有化学相似性的具有成本效益的化合物。我们通过实验设计（DOE）仔细优化了 LLE 条件，考虑了因子相互作用。我们的快速和直接提取和净化方法涉及在五分钟内使用 500 μL 纯甲基叔丁基醚从样品中提取 DPZ。该方法的动态范围为 0.5 ng/mL 至 1,000 ng/mL，显示出优异的灵敏度和适用于不同 DPZ 制剂的药代动力学研究。根据 FDA 指南，我们严格验证了所开发的方法，评估了选择性、线性（相关系数≥0.9999）、准确度（范围为 96.0%至 109.6%）、精密度（≤13.9%）、基质效应（92.2%至 103.8%）、回收率（98.5%至 106.8%）、定量下限（0.5 ng/mL）和稳定性。最后，我们有效地将验证后的方法用于 DPZ 制剂的长期药代动力学评估。我们期望这种方法将对新 DPZ 制剂的发展做出重大贡献，最终使 AD 患者受益。
