Caucasian Gentiana Species: Untargeted LC-MS Metabolic Profiling, Antioxidant and Digestive Enzyme Inhibiting Activity of Six Plants.

The members of Gentiana genus are widely distributed in the Caucasus region where they are used as phytoremedies, but they still have not been studied for their chemical composition and bioactivity. High-performance liquid chromatography with diode array and electrospray triple quadrupole mass detection (HPLC-DAD-ESI-QQQ-MS) was used to investigate metabolites of herb and roots of six gentians (Gentiana asclepiadea, G. cruciata, G. gelida, G. paradoxa, G. pneumonanthe, G. septemfida) grown in the Caucasus. In total, 137 compounds were found including three carbohydrates, 71 iridoid glycosides (mostly loganic acid), loganin, swertiamarin, gentiopicroside and sweroside derivatives, 40 flavones C-, O-, C,O-glycosides (such as luteolin, apigenin, chrysoeriol, and acacetin derivatives), two phenolic O-glycosides, five hydroxycinnamates, eight xanthones, and seven triterpene glycosides. Most of these compounds were identified in gentian samples for the first time. Quantitative differences were found in levels of seven iridoid glycosides, nine glycosylflavones, and two xanthones obtained by HPLC-DAD assay. The gentian extracts were evaluated for their radical-scavenging properties against DPPH and superoxide anion radicals, lipid peroxidation inhibition, and α-amylase/α-glycosidase inhibition. The herb extracts showed higher activity than root extracts. Positive correlations were found between the content of quantified phenolics and antioxidant and digestive enzymes inhibiting activity. The findings presented in our work suggest that the Caucasian gentians are a good source of bioactive phytocompounds with antioxidant and antidiabetic potential.

Caffeoylquinic Acids and Flavonoids of Fringed Sagewort (Artemisia frigida Willd.): HPLC-DAD-ESI-QQQ-MS Profile, HPLC-DAD Quantification, in Vitro Digestion Stability, and Antioxidant Capacity.

Fringed sagewort (Artemisia frigida Willd., Compositae family) is a well-known medicinal plant in Asian medical systems. Fifty-nine hydroxycinnamates and flavonoids have been found in A. frigida herbs of Siberian origin by high-performance liquid chromatography with diode array and electrospray triple quadrupole mass detection (HPLC-DAD-ESI-QQQ-MS). Their structures were determined after mass fragmentation analysis as caffeoylquinic acids, flavone O-/C-glycosides, flavones, and flavonol aglycones. Most of the discovered components were described in A. frigida for the first time. It was shown that flavonoids with different types of substitution have chemotaxonomic significance for species of Artemisia subsection Frigidae (section Absinthium). After HPLC-DAD quantification of 16 major phenolics in 21 Siberian populations of A. frigida and subsequent principal component analysis, we found substantial variation in the selected compounds, suggesting the existence of two geographical groups of A. frigida. The antioxidant activity of A. frigida herbal tea was determined using 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH•) and hydrophilic/lipophilic oxygen radical absorbance capacity (ORAC) assays and DPPH•-HPLC profiling, revealing it to be high. The effect of digestive media on the phenolic profile and antioxidant capacity of A. frigida herbal tea was assessed under simulated gastrointestinal digestion. We found a minor reduction in caffeoylquinic acid content and ORAC values, but remaining levels were satisfactory for antioxidant protection. These results suggest that A. frigida and its food derivate herbal tea could be recommended as new plant antioxidants rich in phenolics.

Khellactone Derivatives and Other Phenolics of Phlojodicarpus sibiricus (Apiaceae): HPLC-DAD-ESI-QQQ-MS/MS and HPLC-UV Profile, and Antiobesity Potential of Dihydrosamidin.

With obesity, the consumption of phenolic-enriched food additives as a part of traditional nutrition avoids the negative implications of eating high-calorie products. This study investigated the new herbal food additive, Phlojodicarpus sibiricus roots and herb, ubiquitously used in Siberia as a spice. Chromatographic techniques such as HPLC-DAD-ESI-QQQ-MS/MS and microcolumn HPLC-UV were the basic instruments for component profiling and quantification, and antiobesity potential was investigated using a differentiated 3T3-L1 adipocytes assay. We found that the roots and herb of P. sibiricus were high-coumarin-containing additives inhibiting triacylglycerol accumulation in 3T3-L1 preadipocytes. Forty-one phenolics were detected in P. sibiricus extracts, and 35 were coumarins, including 27 khellactone derivatives present as esters and glucosides. Total coumarin content varied from 36.16 mg/g of herb to 98.24 mg/g of roots, and from 0.32 mg/mL to 52.91 mg/mL in P. sibiricus preparations. Moreover, Siberian populations of P. sibiricus were characterised by a different HPLC-based coumarin profile. The most pronounced inhibiting effect on triacylglycerol accumulation in 3T3-L1 preadipocytes was shown for dihydrosamidin (khellactone 3'-O-isovaleroyl-4'-O-acetyl ester), which was more active than other khellactone esters and glucosides. The results demonstrated that if used as a food additive Phlojodicarpus sibiricus could be a source of bioactive coumarins of the khellactone group with high antiobesity potential.

Bioactive Phenolics of the Genus Artemisia (Asteraceae): HPLC-DAD-ESI-TQ-MS/MS Profile of the Siberian Species and Their Inhibitory Potential Against α-Amylase and α-Glucosidase.

Artemisia genus of Asteraceae family is a source of medicinal plants known worldwide and used as ethnopharmacological remedies for the treatment of diabetes in Northern Asia (Siberia). The aim of this study was to determine the phenolic profile of 12 Siberian Artemisia species (A. anethifolia, A. commutata, A. desertorum, A. integrifolia, A. latifolia, A. leucophylla, A. macrocephala, A. messerschmidtiana, A. palustris, A. sericea, A. tanacetifolia, A. umbrosa) and to test the efficacy of plant extracts and pure compounds for antidiabetic potential. Finally, by HPLC-DAD-ESI-TQ-MS/MS technique, 112 individual phenolic compounds were detected in Artemisia extracts in a wide range of concentrations. Some species accumulated rare plant phenolics, such as coumarin-hemiterpene ethers (lacarol derivatives) from A. latifolia and A. tanacetifolia; melilotoside from A. tanacetifolia; dihydrochalcones (davidigenin analogs) from A. palustris; chrysoeriol glucosides from A. anethifolia, A. sericea, and A. umbrosa; eriodictyol glycosides from A. messerschmidtiana; and some uncommon flavones and flavonols. The predominant phenolic group from Artemisia species herb was caffeoylquinic acid (CQAs), and in all species, the major CQAs were 5-O-CQA (20.28-127.99 µg/g) and 3,5-di-O-CQA (7.35-243.61 µg/g). In a series of in vitro bioassays, all studied Artemisia extracts showed inhibitory activity against principal enzymes of carbohydrate metabolism, such as α-amylase (IC50 = 150.24-384.14 µg/mL) and α-glucosidase (IC50 = 214.42-754.12 µg/mL). Although many phenolic compounds can be inhibitors, experimental evidence suggests that the CQAs were key to the biological response of Artemisia extracts. Mono-, di- and tri-substituted CQAs were assayed and showed inhibition of α-amylase and α-glucosidase, with IC50 values of 40.57-172.47 µM and 61.08-1240.35 µM, respectively, and they were more effective than acarbose, a well-known enzyme inhibitor. The results obtained in this study reveal that Siberian Artemisia species and CQAs possess a pronounced inhibitory activity against α-amylase and α-glucosidase and could become a complement to synthetic antidiabetic drugs for controlling blood glucose level.

Isorhamnetin and Quercetin Derivatives as Anti-Acetylcholinesterase Principles of Marigold (Calendula officinalis) Flowers and Preparations.

Marigold (Calendula officinalis L.) is one of the most common and widespread plants used medicinally all over the world. The present study aimed to evaluate the anti-acetylcholinesterase activity of marigold flowers, detect the compounds responsible and perform chemical analysis of marigold commercial products. Analysis of 23 varieties of C. officinalis flowers introduced into Siberia allowed us to select the Greenheart Orange variety due to the superior content of flavonoids (46.87 mg/g) and the highest inhibitory activity against acetylcholinesterase (IC50 63.52 µg/mL). Flavonoids, isorhamnetin and quercetin derivatives were revealed as potential inhibitors with the application of high-performance liquid chromatography (HPLC) activity-based profiling. Investigation of the inhibitory activity of isorhamnetin glycosides demonstrated the maximal potency for isorhamnetin-3-О-(2'',6''-di-acetyl)-glucoside (IC50 51.26 µM) and minimal potency for typhaneoside (isorhamnetin-3-O-(2'',6''-di-rhamnosyl)-glucoside; IC50 94.92 µM). Among quercetin derivatives, the most active compound was quercetin-3-О-(2'',6''-di-acetyl)-glucoside (IC50 36.47 µM), and the least active component was manghaslin (quercetin-3-O-(2'',6''-di-rhamnosyl)-glucoside; IC50 94.92 µM). Some structure-activity relationships were discussed. Analysis of commercial marigold formulations revealed a reduced flavonoid content (from 7.18-19.85 mg/g) compared with introduced varieties. Liquid extract was the most enriched preparation, characterized by 3.10 mg/mL of total flavonoid content, and infusion was the least enriched formulation (0.41 mg/mL). The presented results suggest that isorhamnetin and quercetin and its glycosides can be considered as potential anti-acetylcholinesterase agents.

Chemical Composition, Antioxidant and Anticholinesterase Activities of Gentianella azurea from Russian Federation.

Phytochemical study of Gentianella azurea (Bunge) Holub (Gentianaceae) collected in Buryatia Republic (Russian Federation) resulted in the isolation of twenty-one compounds including bellidifolin, bellidin, isobellidifolin, norswertianolin, isobellidifolin-8-O-ß-D-glucopyranoside, orientin, cynaroside, .cosmosiin, apigenin, 4'-O-caffeoylswertiamarin, swertiamarin-6'-O-ß-D-glucopyranoside and sweroside, firstly detected in this species. The extracts and individual compounds were shown to possess antioxidant and anticholinesterase potential.

From medical tradition to traditional medicine: A Tibetan formula in the European framework.

ETHNOPHARMACOLOGICAL RELEVANCE: The increasing prevalence of complex multi-factorial chronic diseases and multimorbidity reveals the need for an enlargement of the therapeutic options. Potent multicompound herbal formulations from traditional medicine systems such as Tibetan Medicine might meet the requirements. With its practice over the centuries Tibetan Medicine is one of the important medical heritages of the world. In the 20th century Tibetan formulas came to Switzerland, where the formula Gabur-25 was then registered as medicine in 1977 (Padma 28, Swissmedic No 35872). The new European directive 2004/24/EC opened the avenue for traditional herbal medicinal products and registrations followed in Austria (HERB-00037) and the UK (39568/0001). The aim of this review was to analyse not only the critical points and hazards but also chances that occur in the endeavour of bringing a ethnopharmacological based preparation to the market within a modern Western medical and regulatory framework and to discuss the necessary transformation steps from a traditional herbal formula towards a modern pharmaceutical product with the example of the Tibetan formula Gabur-25. METHODS: The historic transformation process from the 19th to the 21st century is analysed, using the registration documents and other material from the library of Padma AG, Hinwil, Switzerland. RESULTS: The transformation of a traditional formula into a modern traditional herbal medicinal product according to the present EU regulations is a multi faceted process. The modern indication represents only a small part of the possible traditional indications. Quality and product labelling has to be adopted to modern standards. The formula, once registered, is a fixed combination of herbal and mineral ingredients. Contrary to this the concept of Asian medical tradition allows a certain flexibility in the composition of an herbal formula. The ingredients are constantly adapted to local conditions, availability of raw material and therapeutic situation. CONCLUSIONS: The example shows that in principle complex herbal formulas from Asian medicine can meet the requirements of the European regime of traditional herbal medicinal products. A structured process of transformation from a traditional herbal formula to a modern medicinal product has to include selection of a suitable formula, development of an analytic concept and selection of a suitable indication with regard to the empirical set of possible indications. To extend the range of high quality medicinal products from other medical traditions within the European context the European legislators have to re-evaluate the imposed restrictions given in directive 2004/24/EC. Without amendment of the prerequisite of 15 years documented use in the EU and the limitation of indications for traditional herbal medicinal products, European citizens will be excluded from access to high quality medical traditions with their accumulated empirical knowledge.

[The Tibetan formula Padma Lax in medical practice: a retrospective case series study in constipation including 174 patients]. / Die Tibetische Rezeptur Padma Lax in der ärztlichen Praxis: Eine retrospektive Fallstudie bei Obstipation mit 174 Patienten.

BACKGROUND: The Tibetan Medicine Padma Lax has been approved in Switzerland for the treatment of constipation and bloating for over 40 years. In this study, data on the application in medical practice, as well as on effectiveness in various constipation-related symptoms and on safety and tolerability, were collected and assessed. MATERIAL AND METHODS: Physicians practicing in Switzerland documented retrospectively the course of treatment for chronic constipation, using an especially developed questionnaire. In addition to defecation frequency, data on the intensity of 12 specific symptoms of constipation were collected. RESULTS: 17 participating physicians provided data from a total of 174 patient cases, about 30% of which originated from gynecological practices. Among others, lifestyle factors and various underlying diseases, e.g. neurological or hormonal disturbances, were indicated as causes of the constipation. Safety and tolerability were very good; only 1 adverse event (a worsening of flatulence) was reported. The frequency of defecation increased significantly during treatment and 97% reached a frequency of >2 per week. On average, the frequency and intensity of all symptoms decreased, with the exception of the symptom soft/loose stools, which increased in as many patients as it decreased. The overall intensity of all symptoms decreased from a mean of 15.8 by 63% to 5.8. The proportion of patients with severe complaints (overall intensity score >20) decreased from 20.1% to 1.7%, and as a result, the group with hardly any complaints (overall intensity score ≤ 5) rose from 1.2% to 57%. 86% of the physicians and the patients rated the efficacy as good. CONCLUSIONS: Padma Lax was used in constipation of different causes and in a broad spectrum of patients. Efficacy and safety were assessed as good or very good. In spite of the limitation of the retrospective study design, the data indicate that Padma Lax is an effective and well-tolerated treatment option for chronic constipation from different causes, especially also in gynecological practice, in geriatric patients, and in patients with neurogenic constipation.

Clinical studies on the efficacy and safety of Padma 28, a complex herbal formulation from Tibetan medicine: an overview.

BACKGROUND: Padma 28 is an herbal formula from Tibetan Medicine, which since 35 years has been registered in Switzerland as a drug for the symptoms of circulatory disorders. Over this time, a large body of scientific literature has accumulated. The aim of this article was to give an overview of the clinical studies. METHODS: A systematic literature search was done in PubMed, EMBASE, and the Cochrane Central Register of Controlled Trials (CCRCT). The clinical trials found were assessed regarding fields of application, efficacy, and safety, as well as methodological quality and level of evidence. RESULTS: 29 trials (1 meta-analysis, 21 controlled trials, 7 open trials) and 3 retrospective case studies were found. They deal with different indications and include a total of 1,704 verum (of these, 697 children), 333 placebo, and 394 untreated or healthy patients. Dropouts and withdrawals were 2.5 and 3.5 times higher in the placebo than in the verum group, respectively. The highest level of evidence for the use of Padma 28 was found in the indication of intermittent claudication (11 trials). Indications of efficacy were also found in other vascular (6 trials) and different inflammatory diseases (12 trials). CONCLUSIONS: The results suggest a favorable safety profile for Padma 28, also in the children examined (41% of the study population). Furthermore, the results show a broad field of applications. According to clinical evidence, Padma 28 has shown to be a safe and effective symptomatic treatment option for atherosclerosis-related diseases such as intermittent claudication. It also seems to have a potential for application in certain chronic inflammatory diseases such as recurrent respiratory tract infections, viral hepatitis, and multiple sclerosis. However, further randomized controlled trials (RCT) are needed to confirm these findings.

[Tibetan formulas as pleiotropic signatures--application of network medicines in multimorbidity]. / Tibetische Rezepturen als pleiotrope Signaturen--Einsatz von Netzwerk-Arzneien bei Multimorbidität.

In the context of the network model of the organism, multimorbid states (≥ 2 chronic diseases at the same time) can be considered as a complex disease pattern which can be mapped as characteristic signatures. From the perspective of system theory, living systems such as the human body are viewed as networks of interacting parts. These in turn can themselves be subnetworks assigned to different complexity levels. They range, e.g., from the gene to the transcriptome, proteome, metabolome, and epigenome up to the network of the entire molecular interactions, the so-called interactome. In multimorbidity, the disease signature affects different networks at all levels, e.g., cell systems, organs, and functional systems. Based on this semiotics, certain signatures of effectiveness and profiles of action can be assigned to each drug. A drug signature represents the physicochemical stimuli that cause a reaction by the system, as well as the cross-links by which the entire connected system is affected at all levels. Phytotherapeutics, which chemically represent multi-component mixtures, have especially complex signatures. As multi-target medicines with a pleiotropic effect profile, they therapeutically affect different levels of the network, which is why they are also referred to as network medicines. Herbal formulas from traditional medicine systems such as Tibetan Medicine are an example for phytotherapeutics with a particularly complex pleiotropic signature. Also from the traditional point of view, a disease signature is set in relation with a corresponding drug signature. However, in this case, it is based on the traditional energetic understanding of diseases. Modern research results clearly indicate a widely diversified signature range of Tibetan Medicines and thus provide a rationale for their use in integrative treatment approaches for diseases with complex signatures, e.g. in multimorbidity. The system-theoretical approach discussed here represents a method to enable a connectivity of traditional methods from complementary and alternative medicine to the other disciplines of modern medicine.

Tibetan herbal formula Padma Digestin modulates gastrointestinal motility in vitro.

AIM: To examine the effects of Padma Digestin on the smooth muscle motility of different gastrointestinal segments in vitro. METHODS: The effects of the ethanolic extract of Padma Digestin (at 8.16 mg/mL or 81.6 mg/mL) on the contractility and susceptibility to acetylcholine (ACh) of muscle strips from the cardia, antrum, pylorus, duodenum, jejunum, ileum and colon of male Wistar rats were analyzed. RESULTS: Compared with the control treatment, the Padma Digestin extract had a procontractile effect on the antral smooth muscle strips. Padma Digestin decreased ACh sensitivity in cardia muscle strips and increased it in those from the antrum and pylorus. In the intestinal segments, spontaneous contractility was inhibited in both the duodenal and jejunal strips, whereas reactivity to ACh was inhibited in the jejunal strips only. In the colonic samples, Padma Digestin inhibited spontaneous and ACh-stimulated contractility at a low dose but seems to have increasing effects at a high dose. CONCLUSION: Padma Digestin extract has region-specific effects on the contractility and excitability of gastrointestinal smooth muscle. Our results support the traditional use of Padma Digestin for maldigestion and functional gastrointestinal disorders.

[Early encounters of German-language explorers with the Tibetan medicine in Siberia in the modern era]. / Frühe neuzeitliche Begegnungen deutschsprachiger Forscher mit der tibetischen Medizin in Sibirien.

The spreading of Tibetan Buddhism and with it the Tibetan medicine in the region east of Lake Baikal, goes back to the 17th century. At the beginning of the 18th century, German speaking scholars were among the first to undertake scientific expeditions through Siberia. As such they were amongst the first scientists of the modern era who encountered the traditions, concepts, and therapeutic methods of Tibetan medicine. The aim of this article is to describe and analyze these first encounters with Tibetan medicine by the example of selected men of science of the 18th and 19th century. This work is based on extensive studies of sources in archives and libraries in Russia and Switzerland. We found documents related to the following scientists: Daniel Gottlieb Messerschmidt (1685-1735), Johann Georg Gmelin (1709-1755), Erik Laxmann (1737-1796), Friedrich Adelung (1768-1843), and Joseph Rehmann (1779-1831). They mentioned the distribution of Tibetan medicine within Russia, the use of medicinal plants and formulas as well as therapeutic techniques. For the scientific community of the time these first encounters of Europeans with practitioners of Tibetan medicine could not lift Tibetan medicine out of other exotic context in the field of ethnography. For today's researchers, these encounters are an important evidence for more than 300 years of development of Tibetan medicine on the vast territory of Siberia. The practice and the scientific examination of Tibetan medicine in Siberia is an active endeavor until today. The present work shows that it is possible and rewarding to follow up the historic and cultural connections from Europe to Asia via the Siberian link.

[Anti-inflammatory potential of Padma 28--review of experimental data on the antiatherogenic activity and discussion of the multi-component principle]. / Das anti-inflammatorische Potential von Padma 28--Ubersicht experimenteller Daten zur antiatherogenen Wirkung und Diskussion des Vielstoffkonzepts.

BACKGROUND: The Tibetan remedy Padma 28 has been used in Europe for decades and has proved to be effective in inflammatory and atherosclerotic conditions. Beyond clinical trials, a large number of in vitro and ex vivo studies report various properties and biochemical activities of this complex herbal multicompound. OBJECTIVE: To give an overview of the complex efficacy profile of Padma 28, to review available data, to relate findings to the development of atherosclerosis and thus to discuss the antiatherogenic potential of Padma 28. METHODS: Published non-clinical original papers on Padma 28 were collected and classified according to the studied mechanisms of action. Results were correlated to the briefly described sequences of atherogenesis and various mechanisms of action were elaborated, laying particular emphasis on more recent articles. RESULTS: The complex activity profile of Padma 28 spans mainly direct and indirect anti-inflammatory proper-ties as well as further categories of biochemical actions. These can be related to the complex processes of atherogenesis. CONCLUSIONS: The described mechanisms support the therapeutic field of application of Padma 28, i.e. peripheral circulatory disorders as well as chronic inflammatory disorders. Moreover, the numerous effects as well as the diversity of sites of action allow to draw first conclusions on the conceptual design of this multicomponent formula.