One-pot HPLC method for simultaneous quantification of hydrogen peroxide and organic peroxides

Gabrič, Alen (author), Hodnik, Žiga (author), Ilaš, Janez (author), Pajk, Stane (author)

Hydrogen peroxide and organic peroxides are widely used in the pharmaceutical and food industries as disinfectants, preservatives, and bleaching agents. They can also form as by-products during polymerization, drying, or degradation processes, potentially compromising product quality. While current analytical methods typically measure total peroxide content, they do not distinguish between hydrogen peroxide and organic peroxides, despite their differing reactivity and implications for product stability. We developed a one-pot HPLC method for the simultaneous quantification of hydrogen peroxide and organic peroxides. The method utilizes borinic acid for the selective reaction with hydrogen peroxide, followed by the reaction of triphenylphosphine with the remaining organic peroxides. The resulting phenol derivative and triphenylphosphine oxide are detected using HPLC with UV detection. The method demonstrated strong performance across a variety of complex matrices, including pharmaceutical excipients, milk, honey, and vegetable oil. Quantification limits were 4.4 nmol/g for hydrogen peroxide and 9 nmol/g for organic peroxides. The method's practical utility was demonstrated using famotidine, a pharmaceutical compound selectively oxidized by hydrogen peroxide but not by organic peroxides. This application highlights the method's ability to guide excipient selection based on specific peroxide profiles and to support antioxidant use in formulation development. By enabling the differentiation of peroxide species, this approach facilitates the development of more stable and cost-effective formulations. This novel HPLC method offers a simple, robust, and versatile tool for the selective quantification of hydrogen peroxide and organic peroxides in a wide range of applications, including pharmaceutical and food analysis, where precise peroxide monitoring is essential.

• Language: English
• Keywords: peroxide quantification, hydrogen peroxide, organic peroxides, borinic acid, liquid chromatography, HPLC
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2025
• Number of pages: 8 str.
• Numbering: Vol. 217, art. 114889
• PID: 20.500.12556/RUL-175790
• UDC: 543.426
• ISSN on article: 1095-9149
• DOI: 10.1016/j.microc.2025.114889
• COBISS.SI-ID: 255033859
• Publication date in RUL: 07.11.2025

Record is a part of a journal

• Title: Microchemical journal
• Publisher: Elsevier
• ISSN: 1095-9149
• COBISS.SI-ID: 23088645

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: kvantifikacija peroksida, vodikov peroksid, organski peroksidi, borova kislina, tekočinska kromatografija, HPLC, analizna kemija