Chen Hui, Guo Junping, Zhu Zhenyu, Huang Xiaomei, Guo Jincai
Department of Pharmacy, Changsha Stomatological Hospital, Changsha, China.
School of Stomatology, Hunan University of Chinese Medicine, Changsha, China.
Front Pharmacol. 2025 May 23;16:1592731. doi: 10.3389/fphar.2025.1592731. eCollection 2025.
To develop a rapid, sensitive, and high-throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for prazosin quantification in human plasma, validate its application in bioequivalence studies, and investigate sex-specific pharmacokinetic differences in a Chinese population.
Plasma samples were processed by protein precipitation with methanol and analyzed using a Waters ACQUITY UPLC HSS T3 column. Prazosin-d8 was used as an isotopic internal standard (IS) to enhance quantification accuracy. Chromatographic separation was performed with methanol (A) and 0.1% formic acid in water (B) as the mobile phases, using gradient elution at 0.35 mL/min. Quantification was achieved using positive ionization mode with multiple reaction monitoring (MRM) transitions of m/z 384.2→95.0 for prazosin and m/z 392.2→95.0 for IS.
The method demonstrated excellent linearity (0.1000-30.00 ng/mL, LLOQ: 0.1000 ng/mL), surpassing the sensitivity of prior methods. Bioequivalence analysis confirmed that the 90% confidence interval (CI) for AUC, AUC, and C geometric mean ratios fell within the regulatory acceptance range (90.00%-111.11%). Sex analysis revealed significantly higher AUC (+48%) and AUC (+46%) medians in females (n = 4) than in males (n = 16) ( < 0.05), suggesting potential sex-based differences in prazosin pharmacokinetics.
This study establishes the first LC-MS/MS method integrating isotopic IS and sex-specific pharmacokinetic profiling for prazosin, offering regulatory-compliant bioequivalence validation and insights into precision dosing strategies. These findings support China's generic drug policy and highlight the need for sex-stratified pharmacokinetic evaluations in bioequivalence assessments.
ChiCTR2100050626.
建立一种快速、灵敏且高通量的液相色谱-串联质谱（LC-MS/MS）法用于测定人血浆中哌唑嗪的含量，验证其在生物等效性研究中的应用，并研究中国人群中特定性别的药代动力学差异。
血浆样本采用甲醇蛋白沉淀法处理，并用沃特世ACQUITY UPLC HSS T3柱进行分析。哌唑嗪-d8用作同位素内标（IS）以提高定量准确性。以甲醇（A）和0.1%甲酸水溶液（B）为流动相，采用梯度洗脱，流速为0.35 mL/min进行色谱分离。采用正离子模式和多反应监测（MRM），哌唑嗪的m/z 384.2→95.0和内标的m/z 392.2→95.0进行定量分析。
该方法显示出优异的线性（0.1000 - 30.00 ng/mL，定量下限：0.1000 ng/mL），超过了先前方法的灵敏度。生物等效性分析证实，AUC、AUC和C几何平均比值的90%置信区间（CI）落在监管接受范围内（90.00% - 111.11%）。性别分析显示，女性（n = 4）的AUC中位数（+48%）和AUC中位数（+46%）显著高于男性（n = 16）（P < 0.05），表明哌唑嗪药代动力学可能存在基于性别的差异。
本研究建立了首个用于哌唑嗪的整合同位素内标和特定性别药代动力学分析的LC-MS/MS方法，提供了符合监管要求的生物等效性验证以及对精准给药策略的见解。这些发现支持了中国的仿制药政策，并强调了在生物等效性评估中进行性别分层药代动力学评估的必要性。
ChiCTR2100050626。
