Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist

We developed and validated a liquid chromatography technique to determine organic impurities of Pitolisant in Pitolisant film-coated tablets (4.45 mg and 17.8 mg), a medication used to treat excessive daytime drowsiness. The method uses a gradient elution process on an Inertsil ODS-3 V column (250 m...

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Main Authors: Jayaprakash Kanijam Raghupathi, Lova Gani Raju Bandaru, Kiran Kumar Madineni, Mastan Ali Syed, Sreekantha Babu Jonnalagadda, D. Ramachandran, Naresh Kumar Katari
Format: Article
Language:English
Published: Elsevier 2025-07-01
Series:Results in Chemistry
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Online Access:http://www.sciencedirect.com/science/article/pii/S2211715625003339
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author Jayaprakash Kanijam Raghupathi
Lova Gani Raju Bandaru
Kiran Kumar Madineni
Mastan Ali Syed
Sreekantha Babu Jonnalagadda
D. Ramachandran
Naresh Kumar Katari
author_facet Jayaprakash Kanijam Raghupathi
Lova Gani Raju Bandaru
Kiran Kumar Madineni
Mastan Ali Syed
Sreekantha Babu Jonnalagadda
D. Ramachandran
Naresh Kumar Katari
author_sort Jayaprakash Kanijam Raghupathi
collection DOAJ
description We developed and validated a liquid chromatography technique to determine organic impurities of Pitolisant in Pitolisant film-coated tablets (4.45 mg and 17.8 mg), a medication used to treat excessive daytime drowsiness. The method uses a gradient elution process on an Inertsil ODS-3 V column (250 mm × 4.6 mm, 3 μm), with a flow rate of 0.8 mL/min and detects substances at 210 nm. The analytical parameters consist of a sample injection volume of 10 μL, a sample temperature of 10 °C, and an analytical column compartment temperature of 55 °C. mobile phase A is made up of 70 mM sodium perchlorate monohydrate at a pH of 2.0 mixed with acetonitrile in a 75: 25 ratio, while mobile phase B is a mix of methanol and water in a 90:10 ratio. The recovery rate of impurities ranges from 98.9 % to 104.9 %. All impurities exhibited excellent linearity, with correlation values (r2) of 0.9987 or 0.9999. The method's stability-indicating nature was evaluated through forced degradation studies. Its robustness was assessed using a full factorial design, and we identified the method's operable design space.
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spelling doaj-art-8f815fc063a941749e8aab2faecf60b32025-08-20T03:39:09ZengElsevierResults in Chemistry2211-71562025-07-011610235010.1016/j.rechem.2025.102350Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonistJayaprakash Kanijam Raghupathi0Lova Gani Raju Bandaru1Kiran Kumar Madineni2Mastan Ali Syed3Sreekantha Babu Jonnalagadda4D. Ramachandran5Naresh Kumar Katari6Department of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, India; Department of Food Science, Purdue University, West Lafayette, IN 47906, USADepartment of Chemistry, GITAM School of Science, GITAM Deemed to be University, Hyderabad, Telangana 502329, IndiaDepartment of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, IndiaAnalytical Research & Development, LRF Pharma Pvt Ltd, Hyderabad 500076, IndiaSchool of Chemistry & Physics, College of Agriculture, Engineering & Science, Westville Campus, University of KwaZulu-Natal, P Bag X 54001, Durban 4000, South AfricaDepartment of Chemistry, University College of Sciences, Acharya Nagarjuna University, Nagarjuna Nagar- 522 510, Guntur, Andhra Pradesh, India; Corresponding author.School of Chemistry & Physics, College of Agriculture, Engineering & Science, Westville Campus, University of KwaZulu-Natal, P Bag X 54001, Durban 4000, South Africa; Research & Development, CHEMTEX Environmental Laboratory Inc., Port Arthur, TX, USA; Corresponding author at: School of Chemistry & Physics, University of KwaZulu-Natal, Durban, South Africa.We developed and validated a liquid chromatography technique to determine organic impurities of Pitolisant in Pitolisant film-coated tablets (4.45 mg and 17.8 mg), a medication used to treat excessive daytime drowsiness. The method uses a gradient elution process on an Inertsil ODS-3 V column (250 mm × 4.6 mm, 3 μm), with a flow rate of 0.8 mL/min and detects substances at 210 nm. The analytical parameters consist of a sample injection volume of 10 μL, a sample temperature of 10 °C, and an analytical column compartment temperature of 55 °C. mobile phase A is made up of 70 mM sodium perchlorate monohydrate at a pH of 2.0 mixed with acetonitrile in a 75: 25 ratio, while mobile phase B is a mix of methanol and water in a 90:10 ratio. The recovery rate of impurities ranges from 98.9 % to 104.9 %. All impurities exhibited excellent linearity, with correlation values (r2) of 0.9987 or 0.9999. The method's stability-indicating nature was evaluated through forced degradation studies. Its robustness was assessed using a full factorial design, and we identified the method's operable design space.http://www.sciencedirect.com/science/article/pii/S2211715625003339PitolisantImpuritiesHPLCMethod validationDesign of experimentsFactorial design
spellingShingle Jayaprakash Kanijam Raghupathi
Lova Gani Raju Bandaru
Kiran Kumar Madineni
Mastan Ali Syed
Sreekantha Babu Jonnalagadda
D. Ramachandran
Naresh Kumar Katari
Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist
Results in Chemistry
Pitolisant
Impurities
HPLC
Method validation
Design of experiments
Factorial design
title Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist
title_full Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist
title_fullStr Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist
title_full_unstemmed Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist
title_short Utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in Pitolisant, a Histamine-3 (H3) receptor antagonist/inverse agonist
title_sort utilizing quality by design methodology for liquid chromatography method development and validation in quantifying impurities in pitolisant a histamine 3 h3 receptor antagonist inverse agonist
topic Pitolisant
Impurities
HPLC
Method validation
Design of experiments
Factorial design
url http://www.sciencedirect.com/science/article/pii/S2211715625003339
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