Greenness profile assessment using the on-line SPE-LC method to determine cefuroxime in human breast milk

Abstract

In this study, a pretreatment-free, fully automated on-line SPE-LC method was developed and validated for the quantification of Cefuroxime (CEF) in human breast milk, offering a reliable tool to monitor potential drug transfer to breastfed infants. CEF, a broad-spectrum β-lactam antibiotic, may reach breast milk at low levels; however, according to LactMed, its excretion into human milk is generally minimal and considered unlikely to cause adverse effects in nursing infants. Nevertheless, monitoring its concentration remains clinically valuable, particularly in sensitive populations such as neonates or preterm infants where immature renal clearance may increase susceptibility. Analytical separation was achieved using a C18 analytical column (150 × 4.6 cm, 5 μm) and a self-packed SPE column with a hydrophilic modified strong anion exchange sorbent (20.0 × 1.0 mm). A gradient elution was executed at 1.0 mL/min using acetonitrile and 10 mM o-phosphoric acid solution as the mobile phase detecting CEF at 276 nm. The developed method, which integrates automation and miniaturization principles of green analytical chemistry, eliminates pretreatment steps, reduces solvent consumption, and minimizes waste generation. The method demonstrated satisfactory analytical performance, with a linear range of 0.31–200.00 μg/mL (r = 0.9989), recovery ≥96.52 %, RSD ≤8.84 %, and LOD/LOQ values of 0.10 and 0.31 μg/mL, respectively. The greenness of the method was evaluated using AGREE (score 0.59) and moGAPI (76 %), confirming its environmentally friendly character by enabling precise, eco-friendly, and high-throughput quantification of cefuroxime in breast milk. This method provides a valuable analytical tool to support lactation safety assessments and pharmacovigilance efforts in breastfeeding mothers.