A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study

Background: After surgery, non-depolarizing neuromuscular blocking medications include neostigmine (NEO) and glycopyrrolate (GLY). Numerous traditional approaches, such as HPLC and UPLC procedures, are established for the quantification of GLY and NEO; nevertheless, they lack sensitive and specific...

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Main Authors: P. Vasu Babu, D. Akiladevi
Format: Article
Language:English
Published: Creative Pharma Assent 2025-06-01
Series:Journal of Applied Pharmaceutical Research
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Online Access:https://japtronline.com/index.php/joapr/article/view/834
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author P. Vasu Babu
D. Akiladevi
author_facet P. Vasu Babu
D. Akiladevi
author_sort P. Vasu Babu
collection DOAJ
description Background: After surgery, non-depolarizing neuromuscular blocking medications include neostigmine (NEO) and glycopyrrolate (GLY). Numerous traditional approaches, such as HPLC and UPLC procedures, are established for the quantification of GLY and NEO; nevertheless, they lack sensitive and specific analysis, especially in complex matrices.  Using the LC/MS approach, this work develops a bioanalytical method for quantifying both drugs in rat plasma and applies it to pharmacokinetic studies. Methodology: The plasma was extracted using acetonitrile, and Rivastigmine was employed as an internal standard. An MRM method with positive ions was used for multiple reactions. A  C18 column and a mobile phase - 70:30 mixture of acetonitrile and buffer was utilised at a flow rate of 1 ml/min. Plasma vortex for 10 minutes and centrifuged at 4000 rpm at 20°C. Validation and stability studies are conducted according to the ICH guidelines. The pharmacokinetic study by WinNonlin (Version 5.2) software.  Results and Discussion: Rt for Glycopyrrolate and Neostigmine at 1.838 and 2.800min. GLY has a precision (%CV) of 0.45 at HQC and 3.57 at LQC. NEO had a precision (%CV) of 1.13 at HQC and 2.79 at LQC. From 2 to 40 ng/mL of GLY and 10 to 200 ng/mL of NEO, the standard curves showed a linear relationship. LOD and LOQ for both drugs were 3pg/mL and 10pg/mL. Conclusion: A simple, affordable, reliable, and sensitive approach for quantifying GLY and NEO in rat plasma using LC-MS, with Rivastigmine serving as the internal standard, was developed, validated, and successfully applied in the pharmacokinetic study of rat plasma.
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spelling doaj-art-6cbc3e9e5da94ccab83c836829e494d82025-08-20T04:00:44ZengCreative Pharma AssentJournal of Applied Pharmaceutical Research2348-03352025-06-0113311912810.69857/joapr.v13i3.834835A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic studyP. Vasu Babu0D. Akiladevi1Department of Pharmaceutics, School of Pharmaceutical Sciences, Vels Institute of Sciences, Technology & Advanced Studies, Pallavaram, Chennai-600117, IndiaDepartment of Pharmaceutics, School of Pharmaceutical Sciences, Vels Institute of Sciences, Technology & Advanced Studies, Pallavaram, Chennai-600117, IndiaBackground: After surgery, non-depolarizing neuromuscular blocking medications include neostigmine (NEO) and glycopyrrolate (GLY). Numerous traditional approaches, such as HPLC and UPLC procedures, are established for the quantification of GLY and NEO; nevertheless, they lack sensitive and specific analysis, especially in complex matrices.  Using the LC/MS approach, this work develops a bioanalytical method for quantifying both drugs in rat plasma and applies it to pharmacokinetic studies. Methodology: The plasma was extracted using acetonitrile, and Rivastigmine was employed as an internal standard. An MRM method with positive ions was used for multiple reactions. A  C18 column and a mobile phase - 70:30 mixture of acetonitrile and buffer was utilised at a flow rate of 1 ml/min. Plasma vortex for 10 minutes and centrifuged at 4000 rpm at 20°C. Validation and stability studies are conducted according to the ICH guidelines. The pharmacokinetic study by WinNonlin (Version 5.2) software.  Results and Discussion: Rt for Glycopyrrolate and Neostigmine at 1.838 and 2.800min. GLY has a precision (%CV) of 0.45 at HQC and 3.57 at LQC. NEO had a precision (%CV) of 1.13 at HQC and 2.79 at LQC. From 2 to 40 ng/mL of GLY and 10 to 200 ng/mL of NEO, the standard curves showed a linear relationship. LOD and LOQ for both drugs were 3pg/mL and 10pg/mL. Conclusion: A simple, affordable, reliable, and sensitive approach for quantifying GLY and NEO in rat plasma using LC-MS, with Rivastigmine serving as the internal standard, was developed, validated, and successfully applied in the pharmacokinetic study of rat plasma.https://japtronline.com/index.php/joapr/article/view/834lc-msglycopyrrolateneostigminepharmacokinetics studies
spellingShingle P. Vasu Babu
D. Akiladevi
A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study
Journal of Applied Pharmaceutical Research
lc-ms
glycopyrrolate
neostigmine
pharmacokinetics studies
title A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study
title_full A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study
title_fullStr A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study
title_full_unstemmed A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study
title_short A bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by LC-MS and its application to pharmacokinetic study
title_sort bioanalytical method development and validation for quantification of glycopyrrolate and neostigmine in rat plasma by lc ms and its application to pharmacokinetic study
topic lc-ms
glycopyrrolate
neostigmine
pharmacokinetics studies
url https://japtronline.com/index.php/joapr/article/view/834
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AT pvasubabu bioanalyticalmethoddevelopmentandvalidationforquantificationofglycopyrrolateandneostigmineinratplasmabylcmsanditsapplicationtopharmacokineticstudy
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