Biochemometrics has emerged as promising strategy for the targeted identification of bioactive constituents from natural sources. It is based on the correlation of bioactivity data with chemical data to reveal constituents contributing to activity. Providing complementary data and structural information, MS- and NMR-based biochemometric approaches have both been separately applied in the past. The herein presented study is dedicated to the evaluation of a combined MS- and NMR-based biochemometric workflow for the unambiguous identification of bioactives. As an example, a flower extract of Buddleja officinalis Maxim. was selected to unravel bioactive constituents in the context of dry eye disease pathology. While NMR-based biochemometrics relies on heterocovariance analysis (HetCA) of ¹H NMR spectra using the previously established ELINA approach, a biochemometric molecular network was generated for the MS-based approach. Both analyses were performed in parallel and were ultimately combined to increase their power to identify the bioactive constituents from the complex mixture. As a result, phenylethanoid glycosides and triterpene saponins were discovered as main contributors for the antioxidant and cytotoxic effects of the extract. This article illustrates the advantages, opportunities, and limitations of MS and NMR in the context of biochemometrics.

Dry eye disease (DED) is caused by inflammation on the ocular surface and insufficient quality or production of the tear film. Due to various harmful environmental conditions, a gradual increase of DED cases has been reported. This study aims for a comprehensive in vitro pharmacological and phytochemical profiling of two different Buddleja officinalis Maxim. extracts to assess their potential for the treatment of DED. A hydroethanolic (BO-HE) and a lead-like enhanced (BO-LLE) B. officinalis extract were phytochemically characterized by UHPLC-UV-MS and UHPLC-ELSD analyses. Afterwards, the effects of either BO-HE or BO-LLE on in vitro dry eye models, including human corneal epithelial cell-transformed (HCE-T) cells, immortalized human meibomian gland epithelial cells (IHMGECs) and human leukemia monocytic (THP-1) cells, and Jurkat cells, were investigated. Both extracts exhibited strong anti-inflammatory properties with free radical scavenging activities and reduction of intracellular reactive oxygen species (ROS) in UVB-exposed HCE-T cells. Treatment with BO-HE or BO-LLE showed wound healing capacities. Moreover, both extracts differentially modulate mediator secretion in UVB-exposed HCE-T cells. In IHMGECs, the size of secreted lipid droplets was larger in BO-LLE treated cells. As for immune cells, a significant reduction in levels of TNF-α and IL-6 secretion by lipopolysaccharide (LPS)-exposed THP-1 cells was observed. Additionally, BO-LLE effectively inhibited intracellular calcium influx in Jurkat cells even at low concentrations. The results of this study demonstrate pharmacological potential of B. officinalis flowers for the treatment of DED pathology with BO-LLE exerting a slightly more beneficial profile than BO-HE.

Maca (Lepidium meyenii Walp.), a Brassicaceae species native to the high Andes of Peru, has gained global attention as a functional food and herbal medicinal product due to its endocrine-modulating, fertility-enhancing, and neuroprotective properties. Although numerous studies have addressed its biological effects, a systematic and up-to-date summary of its chemical constituents and analytical methodologies is lacking. This review aims to provide a critical overview of the chemical constituents of L. meyenii and to evaluate analytical studies published between 2000 and 2025, focusing on recent advances in extraction strategies and qualitative and quantitative analytical techniques for quality control. Major compound classes include macamides, macaenes, glucosinolates, and alkaloids, each contributing to maca’s multifaceted activity. Ultra-(high-)performance liquid chromatography (U(H)PLC), often coupled with ultraviolet, diode array, or mass spectrometric detection, is the primary and most robust analytical platform due to its sensitivity, selectivity, and throughput, while ultrasound-assisted extraction improves efficiency and reproducibility. Emerging techniques such as metabolomics and chemometric approaches enhance quality control by enabling holistic, multivariate assessment of complex systems and early detection of variations not captured by traditional univariate methods. As such, they provide complementary, predictive, and more representative insights into maca’s phytochemical complexity. The novelty of this review lies in its integration of conventional targeted analysis with emerging approaches, comprehensive comparison of analytical workflows, and critical discussion of variability related to phenotype, geographic origin, and post-harvest processing. By emphasizing analytical standardization and quality assessment rather than biological activity alone, this review provides a framework for quality control, authentication, and safety evaluation of L. meyenii as a functional food and dietary supplement.

Ethnopharmacological relevanceEthnomedicinal fungi have a long history of use in the treatment of tumors and inflammatory disorders, pathologies often associated with dysregulated angiogenesis and immune-vascular interactions. However, their effects on endothelial cell functions relevant to angiogenesis and inflammation remain insufficiently characterized.Aim of the studyThe aim of this study was to identify the bioactive compounds from ethnopharmacologically relevant medicinal fungi by screening their ethanolic extracts for their effects on angiogenesis- and inflammation-related endothelial cell functions, followed by the identification and targeted isolation of these compounds using molecular networking and bioactivity-guided fractionation.Materials and MethodsTwenty-six ethanolic extracts from traditionally and popularly used macrofungi were screened in vitro for bioactivities related to angiogenesis and inflammation. Assays included endothelial cell proliferation and migration using human umbilical vein endothelial cells (HUVECs), as well as leukocyte adhesion to TNF-activated endothelial cells. Feature-based molecular networking was applied to correlate bioactivity with chemical profiles. Bioactive constituents from Inonotus obliquus were isolated and evaluated for anti-proliferative activity.ResultsEleven fungal extracts significantly inhibited HUVEC proliferation at 25 µg/µL, with Fomitopsis betulina and Inonotus obliquus showing the highest activity. None of the extracts affected endothelial migration, and only one inhibited leukocyte adhesion. Molecular networking revealed a rich and diverse occurrence of lanostane-type triterpenes (LTTs) in all anti-proliferative extracts. Chemical investigation of Inonotus obliquus led to the isolation of several LTTs, of which lanosterol and inotodiol inhibited HUVEC proliferation with IC50 values of 2.1 µM and 9.1 µM, respectively, whereas more oxidized LTTs showed reduced activity.ConclusionsExtracts rich in LTTs exhibit pronounced anti-proliferative effects on endothelial cells, supporting the traditional use of certain polypores in conditions involving aberrant angiogenesis.