A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application

A novel, green, and robust Liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous quantification of sulopenem etzadroxil and probenecid in rat plasma, with application to pharmacokinetic studies. Method development was guided by a Box–Behnke...

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Main Authors: Niloufer Tasnim Khazi, Kumaraswamy Gandla, Lalitha Repudi
Format: Article
Language:English
Published: Elsevier 2025-12-01
Series:Journal of Pharmaceutical and Biomedical Analysis Open
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Online Access:http://www.sciencedirect.com/science/article/pii/S2949771X25000349
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author Niloufer Tasnim Khazi
Kumaraswamy Gandla
Lalitha Repudi
author_facet Niloufer Tasnim Khazi
Kumaraswamy Gandla
Lalitha Repudi
author_sort Niloufer Tasnim Khazi
collection DOAJ
description A novel, green, and robust Liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous quantification of sulopenem etzadroxil and probenecid in rat plasma, with application to pharmacokinetic studies. Method development was guided by a Box–Behnken Design and response surface methodology, optimizing key chromatographic variables—ethanol proportion (40–60 %), flow rate (0.8–1.2 mL/min), and mobile phase pH (2.8–3.2)—to achieve maximum resolution, peak area, and analytical reproducibility. Chromatographic separation was performed on a Kromasil C18 column using ethanol and 0.1 % TFA (50:50, v/v) as the mobile phase. Mass spectrometric detection employed selected reaction monitoring in positive ion mode using an LC-MS/MS instrument. The method exhibited excellent linearity (10–400 ng/mL), low limits of detection (LOD: ∼3 ng/mL), and quantification (LOQ: ∼9 ng/mL) for both analytes, with recovery rates > 93 % and %CVs < 15 %. Greenness and sustainability assessments using analytical GREEnness metric (AGREE), analytical GREEnness metric for sample PREParation (AGREEprep), complementary green analytical procedure index (ComplexGAPI), Eco-Scale, and Blue applicability grade index (BAGI) confirmed the method’s environmental compatibility and analytical reliability, with scores exceeding 65 across all tools. This validated method demonstrates high sensitivity, reproducibility, and environmental responsibility, rendering it suitable for routine bioanalytical and pharmacokinetic applications.
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spelling doaj-art-db7eee327abd4f319149584f0fed01d02025-08-20T02:47:09ZengElsevierJournal of Pharmaceutical and Biomedical Analysis Open2949-771X2025-12-01610008310.1016/j.jpbao.2025.100083A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic applicationNiloufer Tasnim Khazi0Kumaraswamy Gandla1Lalitha Repudi2Department of Pharmacy, Chaitanya (Deemed to be University), Gandipet, Himayathnagar (Vill), Monabad (Mandal), RangaReddy(Dist), Hyderabad, Telangana 500075, IndiaCorresponding author.; Department of Pharmacy, Chaitanya (Deemed to be University), Gandipet, Himayathnagar (Vill), Monabad (Mandal), RangaReddy(Dist), Hyderabad, Telangana 500075, IndiaDepartment of Pharmacy, Chaitanya (Deemed to be University), Gandipet, Himayathnagar (Vill), Monabad (Mandal), RangaReddy(Dist), Hyderabad, Telangana 500075, IndiaA novel, green, and robust Liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the simultaneous quantification of sulopenem etzadroxil and probenecid in rat plasma, with application to pharmacokinetic studies. Method development was guided by a Box–Behnken Design and response surface methodology, optimizing key chromatographic variables—ethanol proportion (40–60 %), flow rate (0.8–1.2 mL/min), and mobile phase pH (2.8–3.2)—to achieve maximum resolution, peak area, and analytical reproducibility. Chromatographic separation was performed on a Kromasil C18 column using ethanol and 0.1 % TFA (50:50, v/v) as the mobile phase. Mass spectrometric detection employed selected reaction monitoring in positive ion mode using an LC-MS/MS instrument. The method exhibited excellent linearity (10–400 ng/mL), low limits of detection (LOD: ∼3 ng/mL), and quantification (LOQ: ∼9 ng/mL) for both analytes, with recovery rates > 93 % and %CVs < 15 %. Greenness and sustainability assessments using analytical GREEnness metric (AGREE), analytical GREEnness metric for sample PREParation (AGREEprep), complementary green analytical procedure index (ComplexGAPI), Eco-Scale, and Blue applicability grade index (BAGI) confirmed the method’s environmental compatibility and analytical reliability, with scores exceeding 65 across all tools. This validated method demonstrates high sensitivity, reproducibility, and environmental responsibility, rendering it suitable for routine bioanalytical and pharmacokinetic applications.http://www.sciencedirect.com/science/article/pii/S2949771X25000349Sulopenem etzadroxilProbenecidMeropenemPharmacokineticsBioanalytical method validationGreen analytical chemistry
spellingShingle Niloufer Tasnim Khazi
Kumaraswamy Gandla
Lalitha Repudi
A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application
Journal of Pharmaceutical and Biomedical Analysis Open
Sulopenem etzadroxil
Probenecid
Meropenem
Pharmacokinetics
Bioanalytical method validation
Green analytical chemistry
title A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application
title_full A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application
title_fullStr A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application
title_full_unstemmed A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application
title_short A green and robust LC–MS/MS bioanalytical method for sulopenem etzadroxil and probenecid: Optimization, validation, and pharmacokinetic application
title_sort green and robust lc ms ms bioanalytical method for sulopenem etzadroxil and probenecid optimization validation and pharmacokinetic application
topic Sulopenem etzadroxil
Probenecid
Meropenem
Pharmacokinetics
Bioanalytical method validation
Green analytical chemistry
url http://www.sciencedirect.com/science/article/pii/S2949771X25000349
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