Terpene transfer to milk from fresh leaves of hemp (Cannabis sativa L.) and savory (Satureja hortensis L.)

ABSTRACT: This study aimed to investigate the transfer of terpenes from fresh aromatic plants, hemp (Cannabis sativa L.) and savory (Satureja hortensis L.), into cow milk, quantify their transfer efficiency, and explore their dynamics. Six Simmental dairy cows were allocated to a single concurrent 3 × 3 Latin square design with 3 dietary treatments: a TMR as control (CTRL) and the same ration supplemented with fresh hemp (HEMP) or savory (SAV) leaves. Each period lasted 14 d, with herb supplementation administered from d 1 to d 6 at increasing doses (0.2–0.8 kg/d DM). Milk samples were collected daily and analyzed to determine terpene concentrations and transfer kinetics. Terpenes were extracted using headspace solid-phase microextraction and analyzed by GC-MS. A total of 24 terpenes were identified across feed and milk. The HEMP had the highest terpene content (85,962 µg/kg), followed by SAV (11,313 µg/kg) and CTRL (228 µg/kg). Corresponding milk concentrations were 388.8 µg/kg (HEMP-diet), 48.7 µg/kg (SAV-diet), and 50.8 µg/kg (CTRL). Despite the lower absolute terpene content in CTRL, its relative carryover to milk was highest (15.4%) compared with HEMP-diet (8.7%) and SAV-diet (6.2%). Key terpenes such as α-farnesene, β-caryophyllene, and p-cymene were transferred to milk. The results demonstrate that terpenes can be transferred to milk in a compound-dependent manner, with hemp producing the highest absolute terpene concentration in milk. This research supports the application of terpene-rich plants in dairy diets to enhance milk quality, traceability, and value, contributing to more sustainable and differentiated dairy production systems.