Optimizing hexane, KOH, and H2O2 methods for lipid removal and organic matter digestion in microplastic analysis of human milk

Zainuddin, Norafiza and Shamsul, Nur Rasyiqah and Ahmad Nasri, Dhea Maisarah and Ismail, Hamizah and Faudzi, Fikriah and Tuan Anuar, Sabiqah and Chinglenthoiba, Chingakham and Ezraneti, Riri (2026) Optimizing hexane, KOH, and H2O2 methods for lipid removal and organic matter digestion in microplastic analysis of human milk. Toxicology and Environmental Health Sciences, 18 (1). pp. 125-133. ISSN 2005-9752 E-ISSN 2233-7784

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Abstract

Objective Microplastics (MPs) contamination is a growing emerging concern with potential health risks, particularly for vulnerable populations such as infants. This study aims to develop and optimize a method for isolating MPs from human breast milk, a complex biological matrix with high lipid and protein content that complicates detection. The goal is to evaluate the effectiveness of a combined chemical extraction approach for isolating MPs and identifying their physical characteristics and polymer types. Methods Breast milk samples were ethically obtained from the Halimatussaadia Mother’s Milk Centre (HMMC). Lipids were removed using hexane followed by potassium hydroxide (KOH) and hydrogen peroxide (H2O2) treatments to digest organic matter. The chemical extraction process was optimized with varying concentrations: hexane (0.5:1, 1:1), 30% H2O2 (1:2, 1:4), and 10% KOH (1:1, 1:3). The treated samples were analysed using Attenuated Total Reflectance Fourier-Transform Infrared (ATR-FTIR) spectroscopy to identify MPs and determine polymer types. Results This study presents an optimized method combining hexane, KOH, and H2O2 to isolate microplastics (MPs) from human breast milk, identifying 12 MPs polymer types, including polyethylene (PE), polypropylene (PP), and polyethylene terephthalate (PET). This approach improves detection accuracy and provides a foundation for future research into potential health risks associated with MPs exposure in infants. However, due to technological limitations, the most prevalent MPs polymer in the samples could not be confirmed. Despite this, the study demonstrated the reliability of the combined hexane-KOH –H2O2 approach for isolating MPs from human milk. Conclusions This study presents an effective and optimized method for isolating MPs from complex biological fluids such as human breast milk. The findings provide a foundation for future research investigating potential health risks associated with MPs exposure in infants, highlighting the need for larger sample sizes and more advanced analytical techniques.

Item Type:	Article (Journal)
Uncontrolled Keywords:	Microplastics · Human breast milk · Lipid extraction · ATR-FTIR spectroscopy · Polymer identification
Subjects:	Q Science > QD Chemistry R Medicine > RA Public aspects of medicine > RA1190 Toxicology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Allied Health Sciences Kulliyyah of Allied Health Sciences > Department of Biomedical Science (Effective:1st July 2011) Kulliyyah of Science Kulliyyah of Science > Institute of Oceanography and Maritime Studies Sultan Ahmad Shah Medical Centre (SASMEC)
Depositing User:	Dr. Norafiza Zainuddin
Date Deposited:	27 Feb 2026 16:38
Last Modified:	27 Feb 2026 16:38
Queue Number:	2026-02-Q2355
URI:	http://irep.iium.edu.my/id/eprint/127078

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