Green chemistry oriented synthesis of p-anisic acid and comparative toxicity versus synthetic product in zebrafish embryos and keratinocytes

Abstract

Abstract p -Anisic acid is an aromatic carboxylic acid used in cosmetic preservation systems. In this study, a trans-anethole-rich essential oil obtained from Pimpinella anisum seeds was converted to p -anisic acid by aqueous potassium permanganate oxidation with acetone as a co-solvent and hydrochloric acid precipitation. Because the route uses a renewable plant-derived feedstock, water as the principal reaction medium, moderate heating, and acetone recovery, it is described as green chemistry-oriented rather than as an intrinsically green process; no comparative life-cycle assessment, process mass intensity, or E-factor calculation was performed. The isolated product was characterized by ¹H and ¹³C nuclear magnetic resonance (NMR) and profiled by gas chromatography-mass spectrometry (GC-MS) against analytical standards. The effects of plant-derived p -anisic acid (NAA) and commercial synthetic p -anisic acid (SAA) on the viability of the HaCaT cells (24 h exposure) and on developmental toxicity in zebrafish embryos (up to 72 hpf) were investigated. In HaCaT cells, NAA caused a smaller concentration-dependent reduction in viability than SAA over 0.0125-0.05% (w/v). NAA preserved igf2 , fabp , vtg , and cyp1a expressions as biomarkers of embryonic development, hepatocellular stress, estrogenic activity, and xenobiotic response, while showing no significant elevation in LPO, NO levels, or GST activity. SAA disrupted oxidant-antioxidant status and the expression of developmental genes. Because both materials contain the same target molecule, the observed differences should be interpreted as batch-specific and potentially related to impurity profiles or other physicochemical attributes rather than as a source-dependent change in the intrinsic properties of p -anisic acid.