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The aerial parts (Herba Saniculae) and the underground parts (Radix Saniculae) of Sanicula europaea (sanicle) have been used traditionally in Austrian folk medicine to treat wounds. Interestingly, in the Austrian vernacular, “Radix Saniculae” can also refer to the underground parts of Cardamine enneaphyllos. This ambiguity can lead to mistakes in using these two plants and, importantly, adulterations. The present work aims to shed light on using Radix Saniculae as a wound-healing agent. Thus, the main components in the aerial and the scarcely investigated underground parts of Sanicula europaea were identified and compared to the underground parts of Cardamine enneaphyllos. For this purpose, different analytical techniques were employed: TLC, HPLC-DAD/ELSD, UHPLC-ESI-MS, and GC-MS. The main components in both Sanicula and Cardamine plant extracts were saccharides. Both parts of Sanicula europaea showed similar compositions: hydroxycinnamic acid derivatives and triterpene saponins. In contrast, the underground parts of Cardamine enneaphyllos contain two glucosinolates and their breakdown products. These findings suggest the same wound-healing activity for the underground parts of Sanicula europaea as was already found for its aerial parts. The glucosinolates detected in Cardamine enneaphyllos substantiate its use in wound healing. Nevertheless, the presented analytical methods allow for easy discovery of adulterations.

The treatment of chronic wounds, an important issue with the growing elderly population, is increasingly hindered by antibiotic resistance. Alternative wound care approaches involve the use of traditional plant-derived remedies, such as purified spruce balm (PSB), with antimicrobial effects and the promotion of cell proliferation. However, spruce balm is difficult to formulate due to its stickiness and high viscosity; dermal products with satisfying technological properties and the scientific literature on this topic are scarce. Thus, the aim of the present work was to develop and rheologically characterize a range of PSB-based dermal formulations with different hydrophilic/lipophilic compositions. Mono- and biphasic semisolid formulations based on different compounds (petrolatum, paraffin oil, wool wax, castor oil, and water) were developed and characterized by their organoleptic and rheological measurements. A chromatographic method of analysis was established, and skin permeation data were collected for pivotal compounds. The results showed that the dynamic viscosity ranged from 10 to 70 Pas at 10/s for the different shear-thinning systems. The best formulation properties were observed for water-free wool wax/castor oil systems with 20% w/w PSB followed by different water-in-oil cream systems. Skin permeation through porcine skin was observed for different PSB compounds (e.g., pinoresinol, dehydroabietic acid, and 15-hydroxy-dehydroabietic acid) using Franz-type diffusion cells. The permeation potential of wool wax/castor oil- and lard-based formulations was shown for all the analyzed substance classes. The varying content of pivotal compounds in different PSB batches collected at different timepoints from different spruce individuals might have contributed to observed differences in vehicle performance.

Picea abies (L.) H. Karst. (Pinaceae) is native to Northern, Central and Eastern Europe. The fast-growing tree reaches up to 50 m in height, has modest nutritional requirements and depends on sufficient water supply. The conifer, commonly called Norway spruce, produces exudates which are traditionally used to treat skin wounds in Northern European countries. Major bioactive constituents of the conifer oleoresin are diterpene resin acids (DRAs) of the abietane and the pimarane type. To assure consistent pharmaceutical quality of Norway spruce balm and commercial products thereof, an analytical method for the quantitation of DRAs is the prerequisite. However, high structural similarity among DRAs and their poor UV absorption makes chromatographic separation and detection challenging: Conventional liquid chromatography systems often fail to achieve sufficient separation, moreover, they are not sustainable. Gas chromatography on the other hand requires time-consuming derivatization prior to unacceptably long analyses (>60 min). These drawbacks prompted the development of the first validated supercritical fluid-based protocol for the separation and quantitation of eight DRAs, i.e., pimaric acid ( 1 ), sandaracopimaric acid ( 2 ), palustric acid ( 3 ), isopimaric acid ( 4 ), levopimaric acid ( 5 ), abietic acid ( 6 ), dehydroabietic acid ( 7 ), and neoabietic acid ( 8 ). By using an ultra high-performance supercritical fluid chromatography (UHPSFC) device hyphenated to a quadrupole mass detector, the DRAs were separated in less than 20 min on a Torus 2-Picolylamin (2-PIC) column (3.0 mm × 100 mm; 1.7 µm particle size) applying supercritical CO 2 and ethanol as mobile phase. Regarding selectivity, accuracy (recovery rates: 87–108%), intermediate precision (between 6.6 and 11.1%), and linearity (R 2 ≥ 0.99; linear between 0.75 μg/ml and 2.5 mg/ml), results were obtained in line with ICH guidelines. The lowest limit of detection (LOD) was at 0.75 μg/ml ( 7 ) and the lowest limit of quantitation (LOQ) at 2 μg/ml ( 8 ). As application examples, 22 Norway spruce balm samples and five commercial products were analyzed. The here presented protocol not only simplifies and shortens the analytical workflow, but also reduces the amount of organic solvent waste by about two thirds compared to conventional liquid chromatographic set-ups. These advantages qualify this fast and efficient method as an ideal tool for an environmentally friendly quality control of traditionally used wound-healing Norway spruce balm products.

Balms and resins of Picea abies, Larix decidua, and Pinus nigra are traditionally used to treat wounds. Three chromatographic techniques differing in separation capacity and technical demands were employed to distinguish among these plant exudates. A TLC method was established for fingerprint comparison, providing a quick overview of a large number of samples at low cost. HPLC-DAD (RP18) and UHPSFC-DAD (Torus 2-Picolylamin), hyphenated to ESI-MS, represented orthogonal chromatographic systems with high separation performance. The developed methods allow for the separation and detection of major and minor constituents belonging to different compound classes (phenyl carboxylic acids, lignans, diterpene resin acids). The qualitative compositions of the diterpene resin acids, the main compounds in the exudates, were comparable in all three genera. Differences were detected in the distribution of hydroxylated diterpene resin acids, pinoresinol, and hydroxycinnamic acids. The three tested chromatographic systems with varying demands on lab equipment offer appropriate tools for the quality assessment of Picea abies, Larix decidua, and Pinus nigra. The extracts were furthermore tested at three different concentrations (10 µg/mL, 3 µg/mL, and 1 µg/mL) for boosted re-epithelialization, a crucial step in the wound-healing process, in an in vitro HaCaT keratinocyte-based scratch assay. Lysophosphatidic acid (LPA, 10 µM) and extracts of several medicinal plants well known for their wound-healing properties (birch, marigold, St. John’s wort, manuka honey) were used as positive controls. Picea abies and Pinus nigra showed concentration dependency; significant activity was measured for Larix decidua at 3 µg/mL.

We describe an unusual case of fulminant tracheobronchial and pulmonary aspergillosis presenting as acute respiratory distress syndrome. The patient, who was apparently immunocompetent, was admitted with severe Plas-modium falciparum malaria but died from aspergillosis.