Elettra | SISSI-Bio Multianalytical investigation reveals psychotropic substances in a Ptolemaic Egyptian vase This multidisciplinary study explores the content of a Ptolemaic Egyptian ceramic “Bes-vase” dated around the 2nd century BCE and preserved at the Tampa Museum of Art, employing a multimodal molecular archaeology approach that integrates synchrotronbased μ-FTIR spectroscopy, proteomics, metabolomics, archaeogenetics, and LC-MS/MS. The objective was to characterize residual organic substances and reconstruct ancient ritual practices involving psychoactive, medicinal, and possibly bodily-derived ingredients. Bes, a popular apotropaic deity in Egyptian religion, was associated with fertility, sexuality, healing, and protective magic, and his image frequently adorned ceramic vessels, commonly referred to as Bes-vases, that were likely used in ritual contexts. Previous hypotheses suggested such vases contained beer, wine, milk, perfumes, or magical mixtures. The examined artifact, classified as type C (Guidotti) and GR1 (Kaiser), shows technological links to molds from El-Fayyūm, confirming its Egyptian provenance and allowing its contextualization within late Hellenistic cultic practices. To investigate the organic residues encrusted inside the vessel, samples were mechanically extracted from the inner surface and analyzed using three complementary techniques. Synchrotron radiation µ-FTIR and ATR-FTIR were carried out at the SISSI beamline of Elettra (Trieste, Italy), enabling identification of chemical substances directly within the ceramic matrix. LC-MS/MS was employed with a tailored metabolomics protocol designed to detect small molecules such as alkaloids, flavonoids, and other plant-derived metabolites. Proteomic analysis targeted ancient peptides, while plant DNA was amplified via trn-intron barcoding to establish botanical origin. The presence of ancient proteins was validated through peptide-spectrum matching and cross-referenced using UniPept, with careful consideration of deamidation rates to rule out modern contamination. Among the most significant findings were alkaloids from Peganum harmala, notably harmaline and harmine, identified through spectral fingerprinting and confirmed via comparison with modern reference samples. This plant is historically known for its psychoactive properties and use in shamanic rituals. Metabolomics and DNA analyses also detected Nymphaea nouchali var. caerulea (blue water lily) and Cleome species, plants associated with sedative, narcotic, or visionary effects. While proteins from these plants were only sporadically preserved, their biochemical signatures were robustly identified. The combined presence of these psychoactive taxa suggests intentional selection for their neurological effects, likely to induce altered states of consciousness. Additionally, proteomic results revealed human proteins, strongly implying the inclusion of bodily fluids in the mixture, a practice that aligns with ritual libation or ingestion and that reinforces the symbolic nature of the contents. Metabolomic data further confirmed the presence of sugars, flavonoids, and organic acids consistent with fermented fruit, likely grape must, and bee-derived products such as honey or royal jelly, as also indicated by μ-FTIR spectra. These ingredients may have served both as flavoring agents and pharmacological carriers for psychoactive compounds. The convergence of psychoactive plants, alcoholic fermentation, bee products, and human-derived material suggests the preparation of a complex psychotropic elixir, possibly consumed or ritually poured during ceremonies aimed at fertility, protection, healing, or divination. This “blood-like” mixture may have been intended to replicate mythological motifs, such as the sedative offering given to Hathor to halt her destructive rampage, and aligns with iconographic and textual sources linking Bes to ecstatic and liminal experiences. The vessel’s function likely extended beyond domestic or medicinal use, entering the domain of trance-inducing rites held in sanctuaries or tombs, such as those attested in the Bes chambers at Saqqara. This research exemplifies the diagnostic power of combined molecular techniques in the study of ancient residues. It reveals the respective strengths and detection limits of FTIR (functional group identification), LC-MS/ MS (low-mass metabolites), proteomics (ancient peptides), and DNA barcoding (botanical source assignment), offering a comprehensive protocol for the analysis of ritual ceramic contents. The ability to detect and cross-validate evidence for psychoactive and symbolic components provides a new methodological framework for the interpretation of similar objects and underscores the importance of integrating multi-omic approaches in archaeological science. The authors suggest that the pattern of ingredients recovered from this Besvase is not an isolated phenomenon but may represent a broader ritual tradition in the Hellenistic-Egyptian world. The implications challenge prior assumptions that Bes-vases served mundane domestic functions and instead position them within the pharmacological and symbolic repertoire of Egyptian sacred practices. In conclusion, the study presents compelling empirical evidence for the preparation and ritual use of a psychoactive and medicinal elixir in Ptolemaic Egypt. The ingredients identified correspond to known psychoactive and symbolic substances in Egyptian tradition, and their combination reflects sophisticated knowledge of pharmacobotany and ritual agency. By reconstructing this complex mixture using state-of-the-art chemical and biological tools, the research advances our understanding of ancient Mediterranean ritual pharmacology and highlights how modern analytical science can illuminate the spiritual and sensory dimensions of past religious experience. Elettra Highlights 2024–2025 56
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