Cannabis is gaining increasing attention due to the high pharmacological potential and updated legislation authorizing multiple uses. The development of time- and cost-efficient analytical methods is of crucial importance for phytocannabinoid profiling. This review aims to capture the versatility of analytical methods for phytocannabinoid profiling of cannabis and cannabis-based products in the past four decades (1980–2021). The thorough overview of more than 220 scientific papers reporting different analytical techniques for phytocannabinoid profiling points out their respective advantages and drawbacks in terms of their complexity, duration, selectivity, sensitivity and robustness for their specific application, along with the most widely used sample preparation strategies. In particular, chromatographic and spectroscopic methods, are presented and discussed. Acquired knowledge of phytocannabinoid profile became extremely relevant and further enhanced chemotaxonomic classification, cultivation set-ups examination, association of medical and adverse health effects with potency and/or interplay of certain phytocannabinoids and other active constituents, quality control (QC), and stability studies, as well as development and harmonization of global quality standards. Further improvement in phytocannabinoid profiling should be focused on untargeted analysis using orthogonal analytical methods, which, joined with cheminformatics approaches for compound identification and MSLs, would lead to the identification of a multitude of new phytocannabinoids.

Background: Wild-growing Cannabis sativa L. represents an underexplored source of hemp-type plant material, but its value for future selection depends on standardized morphological and phytochemical characterization. This study evaluated cultivated Macedonian Cannabis sativa plants of wild-origin (CsW) in comparison with selected commercial cultivars and literature-reported European hemp varieties, based on the assessment of phenotypic differentiation and cannabinoid diversity. Methods: Seeds from five wild-growing plant groups from two N. Macedonian regions, Kočani (W1, W2, W6) and Strumica (W3 and W4), were planted outdoors together with four commercial cultivars with differing cannabinoid composition. Morphological traits were assessed using selected UPOV descriptors and multivariate analysis. Cannabinoids were determined by HPLC-DAD. Results: UPOV-based PCA clearly differentiated CsW from the commercial cultivars (R²X = 0.92, Q² = 0.86), while the expanded comparative UPOV analysis indicated phenotypic affinity with registered hemp-type material, particularly Kompolti, supporting its potential relevance as a phenotypic reference in future DUS-oriented evaluation. Among the cultivated wild-origin groups, W1 from the Kočani region showed the highest proportion of female plants (10:1), representing a favorable agronomic characteristic for flower-oriented production. All CsW were CBD-predominant chemotype III class, with total CBD of 5.14–7.61%, while total Δ9-THC did not exceed 0.20%. This CBD/THC balance positioned the CsW within the higher-CBD range reported for selected European hemp cultivars, while maintaining a favorable low-THC profile. Conclusions: CsW represent morphologically distinct, CBD-rich and low-THC plant material displaying characteristics consistent with a hemp-type profile. The combined UPOV and cannabinoid data support its relevance as a promising resource for further agronomic evaluation, phytochemical selection and future registration-oriented hemp research.

Chronic wounds remain a challenging medical problem due to the involvement of multiple biological processes that can delay normal tissue repair. Medicinal plants may contain diverse bioactive compounds with potential relevance for wound management. However, detailed characterization of their chemical composition is essential for understanding their biological potential. The phytochemical composition of five medicinal plants, Verbena officinalis, Plantago major, Achillea millefolium, Hypericum perforatum, and Calendula officinalis, was investigated using ultra-high-performance liquid chromatography coupled with high-resolution electrospray ionization tandem mass spectrometry (UHPLC-ESI-HRMS/MS). A total of 162 metabolites were identified across the investigated extracts, representing diverse classes including flavonoids, phenolic acids, phenylethanoid glycosides, xanthones, naphthodianthrones, acylphloroglucinols and iridoid glycosides. A comparison with the available literature data and databases confirmed previously reported constituents and revealed 115 unique compounds that had not previously been reported for at least one of the five investigated plant species. The obtained results demonstrate the remarkable chemical diversity of the selected medicinal plants and provide a comprehensive basis for further investigation of their biological activities and potential use in multicomponent herbal mixtures for wound management.

Rutin (a flavonoid glycoside) and verbascoside (a phenylethanoid glycoside) are bioactive phytochemical constituents widely recognized in traditional and modern phytotherapy for their antioxidant, anti-inflammatory and antibacterial activities. The presence of both rutin and verbascoside in a multicomponent herbal mixture (Verbena officinalis L., Hypericum perforatum L., Achillea millefolium L. and Plantago major L.) is expected to enhance its therapeutic efficacy in wound healing. In this study, an HPLC-DAD method was developed and optimized for the simultaneous determination of these two marker compounds within a single analytical run. The method was confirmed for selectivity, specificity, linearity, accuracy, precision, robustness, detection and quantification limits, as well as the stability of the standard mixture. Complete chromatographic resolution (Rs > 1.5) was achieved for both analytes. Excellent linearity (R2 ≥ 0.999) was achieved for both compounds. Accuracy was verified at 50 %, 100 %, and 150 % of the working concentration, while precision was demonstrated by relative standard deviation values below 2 %. Robustness testing showed consistent performance under minor variations in chromatographic conditions. Low detection and quantification limits were obtained for both analytes, and the standard mixture remained stable for up to 24 hours at room temperature. The validated method provides a rapid, reliable, and cost-effective tool for routine quality control and standardization of complex multicomponent herbal formulations.