A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China

ObjectiveTo 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 populati...

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Main Authors: Hui Chen, Junping Guo, Zhenyu Zhu, Xiaomei Huang, Jincai Guo
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-05-01
Series:Frontiers in Pharmacology
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Online Access:https://www.frontiersin.org/articles/10.3389/fphar.2025.1592731/full
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author Hui Chen
Hui Chen
Junping Guo
Zhenyu Zhu
Xiaomei Huang
Jincai Guo
Jincai Guo
author_facet Hui Chen
Hui Chen
Junping Guo
Zhenyu Zhu
Xiaomei Huang
Jincai Guo
Jincai Guo
author_sort Hui Chen
collection DOAJ
description ObjectiveTo 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.Materials and methodsPlasma 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.ResultsThe 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 AUC0-24, AUC0-∞, and Cmax geometric mean ratios fell within the regulatory acceptance range (90.00%–111.11%). Sex analysis revealed significantly higher AUC0-24 (+48%) and AUC0-∞ (+46%) medians in females (n = 4) than in males (n = 16) (P < 0.05), suggesting potential sex-based differences in prazosin pharmacokinetics.ConclusionThis 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.Clinical trial registration numberChiCTR2100050626.
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institution Kabale University
issn 1663-9812
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spelling doaj-art-9e1f6145fbc64c04945c2ef4c3dd08da2025-08-20T03:53:56ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122025-05-011610.3389/fphar.2025.15927311592731A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in ChinaHui Chen0Hui Chen1Junping Guo2Zhenyu Zhu3Xiaomei Huang4Jincai Guo5Jincai Guo6Department of Pharmacy, Changsha Stomatological Hospital, Changsha, ChinaSchool of Stomatology, Hunan University of Chinese Medicine, Changsha, ChinaDepartment of Pharmacy, Hubei Province Corps Hospital, The Chinese Armed Police Force (CAPF), Wuhan, ChinaDepartment of Phase I Clinical Trial Research Center, XiangYa BoAi Rehabilitation Hospital, Changsha, ChinaDepartment of Phase I Clinical Trial Research Center, XiangYa BoAi Rehabilitation Hospital, Changsha, ChinaDepartment of Pharmacy, Changsha Stomatological Hospital, Changsha, ChinaSchool of Stomatology, Hunan University of Chinese Medicine, Changsha, ChinaObjectiveTo 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.Materials and methodsPlasma 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.ResultsThe 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 AUC0-24, AUC0-∞, and Cmax geometric mean ratios fell within the regulatory acceptance range (90.00%–111.11%). Sex analysis revealed significantly higher AUC0-24 (+48%) and AUC0-∞ (+46%) medians in females (n = 4) than in males (n = 16) (P < 0.05), suggesting potential sex-based differences in prazosin pharmacokinetics.ConclusionThis 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.Clinical trial registration numberChiCTR2100050626.https://www.frontiersin.org/articles/10.3389/fphar.2025.1592731/fullprazosinprazosin-d8LC-MS/MSpharmacokineticsbioequivalence
spellingShingle Hui Chen
Hui Chen
Junping Guo
Zhenyu Zhu
Xiaomei Huang
Jincai Guo
Jincai Guo
A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China
Frontiers in Pharmacology
prazosin
prazosin-d8
LC-MS/MS
pharmacokinetics
bioequivalence
title A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China
title_full A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China
title_fullStr A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China
title_full_unstemmed A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China
title_short A sex-sensitive LC-MS/MS method with isotopic internal standard for prazosin bioequivalence: bridging precision medicine and generic drug policy in China
title_sort sex sensitive lc ms ms method with isotopic internal standard for prazosin bioequivalence bridging precision medicine and generic drug policy in china
topic prazosin
prazosin-d8
LC-MS/MS
pharmacokinetics
bioequivalence
url https://www.frontiersin.org/articles/10.3389/fphar.2025.1592731/full
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