Prenatal Developmental Evaluation of Phaleria macrocarpa Kombucha: An Experimental Study in Pregnant Mice with GC–MS-Based Metabolite Profiling

Abstract

Kombucha is a fermented beverage increasingly consumed for its reported functional properties, yet information on its developmental effects during pregnancy remains limited. This exploratory study evaluated prenatal developmental outcomes following exposure to Phaleria macrocarpa fruit infusion and kombucha during organogenesis in mice and characterized the fermented preparation using GC–MS. Twenty pregnant mice were allocated to four groups (n = 5 litters per group) and treated orally from gestation day (GD) 6 to GD 15. Maternal body weight, litter size, fetal viability, fetal body weight, crown–rump length, and external, visceral, and skeletal findings were assessed on GD 18. Maternal body-weight gain did not differ significantly among groups. Mean litter size was lower in the treated groups than in the control group, while fetal deaths were infrequent and occurred only in kombucha-treated litters. Mean fetal body weight and crown–rump length were significantly lower in treated groups, with the lowest values in the higher-exposure kombucha group. No gross external, visceral, or skeletal malformations were identified among the fetuses examined. GC–MS profiling tentatively identified several volatile and semi-volatile constituents, including phenolic derivatives, alcohol-related compounds, fatty-acid-related constituents, and 5-hydroxymethylfurfural. Given the limited number of litters and restricted maternal and reproductive assessments, these findings should be interpreted as preliminary growth-associated observations rather than definitive evidence of developmental toxicity or safety. Larger, adequately powered studies with comprehensive reproductive endpoints and quantitative chemical characterization are warranted.