Comparative Phytoprofiling of Achillea millefolium Morphotypes: Assessing Antioxidant Activity, Phenolic and Triterpenic Compounds Variation across Different Plant Parts.

Achillea millefolium L., commonly known as yarrow, is a versatile and widely distributed plant species with a rich history of ethnopharmacological significance. This study aimed to evaluate the comparative differences of A. millefolium inflorescence morphotypes. The phytochemical profile of white and pink inflorescence morphotypes was characterised by a complex of thirty-four phenolic and triterpene compounds. The species has distinct morphotypes of white and pink inflorescence. Phenolic and triterpenic profiles were determined, and individual compounds were quantified in inflorescence, leaf, and stem samples of two morphotypes tested. The antioxidant activity of plant extracts was evaluated by free radical scavenging (ABTS) and ferric-reducing antioxidant power (FRAP) assays. Caffeoylquinic acids predominated in all parts of the plant tested. Chlorogenic acid and 3,5-dicaffeoylquinic acid were the principal compounds in the phenolic profile. Betulin, betulinic acid, and α-amyrin were the prevailing triterpenic components in the triterpenic profiles of Achillea millefolium morphotypes. The predominant flavonoids in inflorescences were flavones, while in leaves, flavonols were the organ-specific compounds. The quantitative differences were observed between plant parts of morphotypes. Leaves consistently displayed the highest amounts of identified compounds and have been testified as the main source of antioxidant activity. Overall, white inflorescences accumulated a higher total amount of compounds compared to pink ones. The observed differences between morphotypes derived from the same population reflect the differences in specialised metabolites and their chemotypes. This study addresses gaps in knowledge, particularly in phenolic and triterpenic profiling of coloured inflorescence morphotypes, enhancing our understanding of chemotypes and morphotypes within the species.

Comparative Analysis of Root Phenolic Profiles and Antioxidant Activity of Five Native and Invasive Solidago L. Species.

The high environmental importance of invasive goldenrod has prompted research to find potential benefits that can be derived from these species. This study aimed to identify differences in root phenolic profiles among five Solidago species, thus providing valuable information on their potential applications and the botanical origin of the raw material. The roots of native S. virgaurea L., two alien species S. gigantea Aiton and S. canadensis L. and their hybrids S. ×niederederi Khek and S. ×snarskisii Gudz. & Zaln. were sampled from mixed-species stands in Lithuania. A complex of twelve phenolic acids and their derivatives was identified and quantified in methanol-water root extracts using the HPLC-PDA and LC/MS systems. The radical-scavenging capacities of the extracts were assessed by ABTS. The chemical content of the roots of S. virgaurea, S. gigantea and S. ×niederederi were statistically similar, while the roots of S. canadensis and S. ×snarskisii contained lower amounts of compounds than the other species. The PCA score-plot models of the phenolic profiles only partially confirmed the identification of S. ×niederederi and S. ×snarskisii as crosses between native and alien species. The findings from the phenolic profiles and the observed radical-scavenging activity of root extracts of Solidago species provide valuable insights into their potential applications in various fields.

Trends in Phenolic Profiles of Achillea millefolium from Different Geographical Gradients.

The traditional widely used raw material of Achillea millefolium is currently mainly derived from wild populations, leading to diversification and uncertainty in its quality. The aim of the study was to determine the accumulation differences of phenolic compounds between geographically distant populations of Achillea millefolium from northern and southern gradients. Plant material was collected from Gaziantep and Nevsehir provinces in Turkey and from wild populations in Lithuania. A complex of nine hydroxycinnamic acids and eleven flavonoids was identified and quantified in the methanolic extracts of inflorescences, leaves, and stems using the HPLC-PDA method. Caffeoylquinic acids predominated in leaves, while inflorescences tended to prevail in flavonoids. The PCA score plot model represented the quantitative distribution pattern of phenolic compounds along a geographical gradient of populations. The content of phenolic compounds in plant materials from northern latitudes was more than twice that of plants from southern latitudes. A significant correlation of individual phenolic compounds with latitude/longitude corresponded to their differences between two countries. Differences in accumulation of caffeoylquinic acids and flavonoids revealed several intraspecific groups within A. millefolium. Our findings suggest that spatial geographical data on the distribution of phenolic compounds in A. millefolium populations could be used as a tool to find potential collection sites for high-quality raw materials.

Distribution Patterns of Essential Oil Terpenes in Native and Invasive Solidago Species and Their Comparative Assessment.

The importance of invasive Solidago L. species to the environment creates a new approach to controlling their spread through the use of potentially high value raw materials. The aim of this study was to assess the distribution patterns of volatile compounds in the four Solidago spp., by identifying common and species-specific compounds with their potentials, and to confirm the origin of the spontaneous hybrid Solidago × niederederi on the basis of comparative assessment of essential oil (EO) profiles. Plant material in the flowering phase was collected in mixed populations from six different sites. The EOs were isolated separately from the leaf and the inflorescence samples by hydrodistillation for 3 h. The chemical analysis was performed by gas chromatography-mass spectrometry. Multivariate data analysis was employed to explain the interspecies relationships among Solidago spp. The results revealed the similarity among Solidago spp. EO profiles, which were dominated by monoterpenes and oxygenated compound fractions. Solidago spp. differed in species distinctive terpenes and their distribution between accessions and plant parts. Volatile compound patterns confirmed the origin of Solidago × niederederi between Solidago canadensis and Solidago virgaurea, with the higher contribution of alien species than native ones. Correct taxonomic identification of species is highly essential for the targeted collection of raw material from the wild for different applications. Solidago spp. can be considered to be underutilized sources of bioactive secondary metabolites.

Distribution of Phenolic Compounds and Antioxidant Activity in Plant Parts and Populations of Seven Underutilized Wild Achillea Species.

Evaluation of phytochemical composition of underutilized Achillea species provides the primary selection of germplasms with the desired quality of raw material for their further applications. The aim of the study was to evaluate the comprehensive distribution patterns of phenolic compounds in seven wild Achillea spp. and their plant parts, and to assess their antioxidant activity. Plant material was collected from different sites in Turkey. A complex of hydroxycinnamic acids, flavonols and flavones was identified and quantified in methanolic extracts using HPLC-PDA method. Antioxidant activity was assessed by radical scavenging assay. The results showed that qualitative and qualitative profiles of caffeoylquinic acids and flavonoids were species-specific, explaining the characteristic patterns of their variation in the corresponding species and plant parts. The highest total amount of caffeoylquinic acids was detected in A. setacea. A. arabica exposed the highest accumulation of mono-caffeoylquinic acids and flavonoids with the greatest levels of quercetin and luteolin derivatives and the flavonol santin. Santin was detected in all plant parts of A. cappadocica, A. setacea, A. santolinoides subsp. wilhelmsii, and A. arabica. A notable antiradical capacity was confirmed in A. arabica, A. setacea and A. cappadocica plant extracts. The leaves of all studied species were found to have priority over inflorescences and stems in terms of radical scavenging activity. The new data complemented the information that may be relevant for the continuation of chemophenetic studies in the heterogeneous genus Achillea.

Factors affecting the variation of bioactive compounds in Hypericum species.

The genus Hypericum (Hypericaceae) consists of 484 species from 36 sections with worldwide distribution in different areas. Turkey is considered as hot spot for diversity of Hypericum genus. Despite numerous publications, Hypericum species still attracted considerable scientific interest due to pharmaceutically relevant secondary metabolites: naphthodianthrones, acylphloroglucinol derivatives, phenolic acids, flavonoid glycosides, biflavonoids, and some other valuable constituents. Phytochemical investigations carried out on different Hypericum species provided highly heterogeneous results. The content of bioactive compounds varies significantly due to many internal and external factors, including plant organs, phenological stage, genetic profile, environmental abiotic and biotic factors, such as growing site, light, temperature, radiation, soil drought and salinity, pathogens, and herbivores attack. The variations in content of bioactive compounds in plants are regarded as the main problem in the standardization of Hypericum-derived pharmaceuticals and dietary supplements. The review discusses the main factors contributing to the variations of bioactive compounds and what kind of modulations can increase quality of Hypericum raw material.

Secondary metabolites of seven Hypericum species growing in Turkey.

Context The genus Hypericum (Hypericaceae) has attracted remarkable scientific interest as its members have yielded many bioactive compounds. Objective The current study presents investigations on the accumulation of hypericin, pseudohypericin, hyperforin, adhyperforin, chlorogenic acid, neochlorogenic acid, caffeic acid, 2,4-dihydroxybenzoic acid, 13,118-biapigenin, hyperoside, isoquercitrin, quercitrin, quercetin, avicularin, rutin, (+)-catechin and (-)-epicatechin in seven Hypericum (Hypericaceae) species growing wild in Turkey, namely, H. aviculariifolium Jaup. and Spach subsp. aviculariifolium (Freyn and Bornm.) Robson var. albiflorum (endemic), H. bithynicum Boiss., H. calycinum L., H. cardiophyllum Boiss., H. elongatum L. subsp. microcalycinum (Boiss. and Heldr.) Robson, H. hirsutum L. and H. xylosteifolium (Spach) N. Robson. Materials and methods The plant materials were collected at flowering period and dissected in different tissues. Air-dried plant material including stems, leaves and flowers was mechanically powdered with a laboratory mill and samples (0.1 g) were extracted in 10 mL of 100% methanol by ultrasonication at 40 °C for 30 min for HPLC-PDA analyses. Results Accumulation levels of the investigated compounds varied greatly depending on species and plant part. Discussion For the first time, the detailed chemical profiles of corresponding Turkish Hypericum species were reported and the results were discussed from a phytochemical point of view. Conclusions The present data have importance in evaluation of plant resources of Hypericum genus in selecting the new potential sources of bioactive compounds.

Development of an HPLC post-column antioxidant assay for Solidago canadensis radical scavengers.

The aim of this work was to modify and validate the post-column high-performance liquid chromatography (HPLC)-ABTS and DPPH methods for evaluating the antioxidant activity of the methanolic extracts of Solidago canadensis (Canadian goldenrod) leaves and flowers. Separation of the analytes was performed via the HPLC-PDA method on a YMC analytical column using a gradient elution program. Three compounds with antioxidant properties - chlorogenic acid, rutin and isoquercitrin - and two unidentified antioxidants were established. The research showed that the coil temperature regimes and loop length combinations influence the optimised post-column assay method for detecting the antioxidant activity of goldenrod radical scavengers. Investigations established that the temperature in the reaction coil was a substantial factor contributing to the signal strength of the analytes after reacting with the DPPH and ABTS radicals.

Population Variability of Main Secondary Metabolites in Hypericum lydium Boiss. (Hypericaceae).

In the present study, we investigated the variation in the content of naphthodianthrones hypericin and pseudohypericin, phloroglucinol derivatives hyperforin and adhyperforin, the phenolic acids as chlorogenic acid, neochlorogenic acid, 2,4-dihydroxybenzoic acid, and the flavonols, namely, hyperoside, isoquercitrin, quercitrin, quercetin, avicularin, rutin, (+)-catechin and (-)-epicatechin, and biflavonoid amentoflavone among wild H. lydium Boiss. populations from five different growing sites of Turkey for the first time. The aerial parts representing a total of 30 individuals were collected at full flowering and dissected into floral, leaf and stem tissues. After dried at room temperature, the plant materials were assayed for chemical contents by HPLC. The populations varied significantly in the content of chemical compounds. Among different plant parts, flowers were found to be main repository site of hyperforin, adhyperforin, hypericin, pseudohypericin, amentoflavone, quercetin, avicularin, rutin and (+)-catechin accumulations whereas rest of the compounds tested were accumulated primarily in leaves in all growing localities. The stems were the least accumulative organ that did not yield hyperforin, adhyperforin and rutin. The chemical diversity among the populations and plant parts is discussed as being possibly the result of different environmental, morphological and genetic factors.

Changes in the content of bioactive substances among Hypericum montbretii populations from Turkey

In the present study, we investigated the variation in the content of seventeen secondary metabolites among Hypericum montbretii Spach., Hypericaceae, populations from five different growing zones in Turkey for the first time.The plants were collected at full flowering, and after they were dried at room temperature, they were assayed for chemical contents by HPLC. Chemical constituents of plants varied significantly among populations except for 2,4-dihydroxybenzoic acid which was accumulated at similar levels. Plants from population - 1 yielded the highest amount of hypericin and pseudohypericin (1.27 and 2.97 mg/g, respectively) while hyperforin and adhyperforin accumulations were the highest in plants from population - 2 (6.64 and 1.24 mg/g, respectively). (+)-Catechin and (-)-epicatechin were accumulated at significantly higher levels by plants of population - 4 (1.54 and 4.35 mg/g, respectively). The highest accumulation level of the rest compounds namely, chlorogenic and neochlorogenic acids, amentoflavone, hyperoside, isoquercitrin, quercitrin, quercetin, avicularin and rutin was reached in plants from population-5 (2.64, 4.37, 2.35, 10.26, 3.52, 4.37, 1.55, 1.56 and 20.54 mg/g, respectively). The pronounced chemical diversity between populations is discussed to possibly be the result of different environmental, morphological and genetic factors.

Changes in the contents of main secondary metabolites in two Turkish Hypericum species during plant development.

CONTEXT: The genus Hypericum (Guttiferae) has received considerable scientific interest as a source of biologically active compounds. OBJECTIVE: The study determined the morphogenetic and ontogenetic variation in the main bioactive compounds of two Hypericum species, namely, Hypericum aviculariifolium subsp. depilatum var. depilatum (Freyn and Bornm.) Robson var. depilatum and Hypericum orientale L. through HPLC analyses of whole plants as well as individual plant parts (stems, leaves, and reproductive tissues). MATERIALS AND METHODS: The plant materials were harvested at five phenological stages: vegetative, floral budding, full flowering, fresh fruiting, and mature fruiting; dried at room temperature, then assayed for chemical content. RESULTS: In H. aviculariifolium, no kaempferol accumulation was observed and the highest level of hypericin, pseudohypericin, and quercitrin was reached at full flowering (0.71, 1.78, and 4.15 mg/g DW, respectively). Plants, harvested at floral budding produced the highest amount of rutin, hyperoside, and isoquercitrine (32.96, 2.42, 1.52 mg/g DW, respectively). H. orientale did not produce hypericin, pseudohypericin, or kaempferol. Rutin, hyperoside, and isoquercetine levels were the highest at floral development (1.76, 11.85, and 1.21 mg/g DW, respectively) and plants harvested at fresh fruiting produced the highest amount of quercitrine and quercetine (0.20 and 1.30 mg/g DW, respectively). DISCUSSION: For the first time, the chemical composition of the Turkish species of Hypericum was monitored during the course of ontogenesis to determine the ontogenetic and morphogenetic changes in chemical content. CONCLUSIONS: Plant material should be harvested during flower ontogenesis for medicinal purposes in which the content of many bioactive substances tested reached their highest level.

Secondary metabolites of Hypericum leptophyllum Hochst., an endemic Turkish species.

In the present study, the presence of the phloroglucinol derivative hyperforin, the naphthodianthrones hypericin and pseudohypericin, the phenylpropane chlorogenic acid and the flavonoids rutin, hyperoside, kaempferol, isoquercetine, quercitrine, and quercetine was investigated in Hypericum leptophyllum Hochst., an endemic Turkish species for the first time. The aerial parts representing a total of 30 individuals were collected at full flowering and dissected into floral, leaf, and stem tissues. After being dried at room temperature, the plant materials were assayed for secondary metabolite concentrations by HPLC. Aerial plant parts accumulated chlorogenic acid, hyperoside, isoquercetine, quercitrine, and quercetine, but they did not accumulate hyperforin, hypericin, pseudohypericin, rutin, and kaempferol. Accumulation levels of the detected compounds varied with plant tissues. Such kind of data could be useful for elucidation of the chemotaxonomical significance of the corresponding compounds and phytochemical evaluation of this endemic species.

Secondary metabolites of Hypericum confertum and their possible chemotaxonomic significance.

The phloroglucinol derivative hyperforin, the naphthodianthrones hypericin and pseudohypericin, the phenylpropane chlorogenic acid, and the flavonoids rutin, hyperoside, apigenin-7-O-glucoside, kaempferol, quercitrin, quercetin and amentoflavone were investigated in Hypericum confertum growing wild in Turkey. After drying at room temperature, the plant materials were assayed for secondary metabolite concentrations by HPLC. All the listed compounds were detected at various levels. This is the first report on the chemistry of H. confertum.

The quantitative effects of temperature and light intensity on phenolics accumulation in St. John's wort (Hypericum perforatum).

The quantitative effects of temperature and light intensity on accumulation of phenolics were examined on greenhouse-grown plants of Hypericum perforatum L. Plants were grown in a greenhouse separated into two parts: shaded by 50% transparent polyethylene cover and un-shaded. Temperature values and light intensities were measured daily during the experiment, while plants were harvested weekly for HPLC analyses. Multi regression analyses were performed to describe the quantitative effects of temperature and light intensity on phenolics accumulation. According to the results, increases in temperatures from 24 degrees C to 32 degrees C and light intensities from 803.4 microMm(-2)s(-1) to 1618.6 microMm(-2)s(-1) resulted in a continuous increase in amentoflavone, apigenin-7-glucoside, cholorogenic acid, hyperoside, kaempferol, rutin, quercetin and quercitrin contents. The relationships between temperature, light intensity and phenolics accumulation were formulized as P= [a + (b1 x t) + (b2 x l) + [b3 x(t x l)]] equition, where P is the content of the corresponding phenolic, t temperature (degrees C), l light intensity (microMm(-2)s(-1)) and a, b1, b2 and b3 the coefficients of the produced equation. The regression coefficient (R2) value for amentoflavone was 0.84, for apigenin-7-glucoside 0.87, for cholorogenic acid 0.83, for hyperoside 0.95, for kaempferol 0.76, for rutin 0.70, for quercetin - 0.93, and for quercitrin - 0.86. All R2 values and standard errors of the equations were found to be significant at the p<0.001 level. The mathematical models produced in the present study could be applied by Hypericum researchers as useful tools for the prediction of phenolics content instead of routine chemical analyses.

Chemical constituents of Hypericum adenotrichum Spach, an endemic Turkish species.

The present study was conducted out to determine hyperforin, hypericin, pseudohypericin, chlorogenic acid, rutin, hyperoside, quercitrin, quercetin, kaempferol, apigenin-7-O-glucoside and amentoflavone contents of Hypericum adenotrichum, an endemic plant species to Turkey. The aerial parts representing a total of 30 individuals were collected at full flowering, dried at room temperature and assayed for secondary metabolite concentrations by HPLC. All of the chemicals were detected at various levels except for hyperforin. This is the first report on polar chemistry of this endemic species.

Variability of phenolic compounds in flowers of Achillea millefolium wild populations in Lithuania.

Achillea millefolium L. sensu lato (yarrow) is the best-known species of the genus Achillea due to numerous medicinal applications both in folk and conventional medicine. Phenolic compounds such as flavonoids and phenol carbonic acids are present in yarrow and constitute one of the most important groups of pharmacologically active substances. In the present study, yarrow flowers gathered from native populations in different locations of Lithuania were analyzed for phenolic compound composition. High-performance liquid chromatography (HPLC) was used for chemical analyses. Eight phenolic compounds--chlorogenic acid and flavonoids, namely vicenin-2, luteolin-3',7-di-O-glucoside, luteolin-7-O-glucoside, rutin, apigenin-7-O-glucoside, luteolin, and apigenin--were identified in the extracts from yarrow flowers. Considerable variation in accumulation of phenolic compounds among the flowers from different locations was observed. The samples were divided into two main groups based on chemical composition: the first group was characterized by lower than the mean total amount of the identified phenolics; the second was formed from samples accumulating higher concentrations of investigated secondary metabolites. The total amount of the identified phenolics in yarrow flowers from different populations varied from 13.290 to 27.947 mg/g.

Pseudohypericin and hyperforin in Hypericum perforatum from Northern Turkey: variation among populations, plant parts and phenological stages.

Hypericum perforatum is a perennial medicinal plant known as "St. John's wort" in Western Europe and has been used in the treatment of several diseases for centuries. In the present study, morphologic, phenologic and population variability in pseudohypericin and hyperforin concentrations among H. perforatum populations from Northern Turkey was investigated for the first time. The aerial parts of H. perforatum plants representing a total of 30 individuals were collected at full flowering from 10 sites of Northern Turkey to search the regional variation in the secondary metabolite concentrations. For morphologic and phenologic sampling, plants from one site were gathered in five phenological stages: vegetative, floral budding, full flowering, fresh fruiting and mature fruiting. The plant materials were air-dried at room temperature and subsequently assayed for chemical concentrations by high performance liquid chromatography. Secondary metabolite concentrations ranged from traces to 2.94 mg/g dry weight (DW) for pseudohypericin and traces -6.29 mg/g DW for hyperforin. The differences in the secondary metabolite concentrations among populations of H. perforatum were found to be significant. The populations varied greatly in hyperforin concentrations, whereas they produced a similar amount of pseudohypericin. Concentrations of both secondary metabolites in all tissues increased with advancing of plant development and higher accumulation levels were reached at flowering. Among different tissues, full opened flowers were found to be superior to stems, leaves and the other reproductive parts with regard to pseudohypericin and hyperforin accumulations. The present findings might be useful to optimize the processing methodology of wild-harvested plant material and obtain increased concentrations of these secondary metabolites.

Evaluation of phenolic acids and phenylpropanoids in the crude drugs.

Phenolic acids and phenylpropanoids have an important biological activity and are therapeutic agents of crude drugs. Development of validated analysis techniques of these phytotherapeutic agents (fingerprinting and assay procedures) is an important practice for efficacy, safety, and quality control of herbal drug preparations. The aim of the present work was to study analytical capabilities of the evaluation of selected phenolic acids and phenylpropanoids: caffeic acid, chlorogenic acid, cinnamic acid, coumaric acid, ferulic acid, gallic acid, protocatechuic (3,4-dihydroxybenzoic) acid, rosmarinic acid, vanillic acid, and vanillin. Optimization and validation procedures of rapid and simple method of reversed-phase high-performance liquid chromatography were carried out. The mobile phase of the optimized chromatographic method consisted of methanol and 0.5% acetic acid solvent in water. For the application of method, two kinds of raw materials were chosen: propolis and the Herba Origani. Coumaric acid is the dominating phenolic acid of propolis (2785 microg/g). Results of analysis of Herba Origani demonstrated high quantities (6376 microg/g) of rosmarinic and protocatechuic (1485 microg/g) acids in the samples.

Variation of bioactive substances in Hypericum montbretii during plant growth.

The present study was conducted to determine phenologic and morphogenetic variation of chlorogenic acid and flavonoids, as rutin, hyperoside, apigenin-7-O-glucoside, quercitrin, quercetin and viteksin content of Hypericum montbretii growing in Turkey. Wild growing plants were harvested at vegetative, floral budding, full flowering, fresh fruiting and mature fruiting stages and dissected into stem, leaf and reproductive tissues and assayed for bioactive compounds by HPLC method. Accumulation of rutin and quercetin was not detected in plant parts of H. montbretii during plant growth. Chlorogenic acid and hyperoside content in whole plant was decreased linearly with advancing of development stages and reached their highest level at vegetative stage. On the contrary, apigenin-7-O-glucoside, quercitrin and viteksin content in whole plant increased during the course of seasonal development and the highest level of those compounds was observed at the stage of full flowering. Leaves did not produce apigenin-7-O-glucoside, while viteksin was not detectable in stem and reproductive tissues. Depending on development stages, reproductive parts had the highest level of apigenin-7-O-glucoside and leaves produced major amount of chlorogenic acid, hyperoside and viteksin whereas accumulation of quercitrin was prevailed in stem tissue. Such kind of data could be useful for elucidation of the chemotaxonomical significance of these compounds and medicinal evaluation of this species.

[Improvement of diversity, trade and conservation of medicinal and aromatic plants]. / Vaistiniu ir aromatiniu augalu ivairoves tyrimo, ju panaudojimo bei issaugojimo tendencijos.

The significance of medicinal and aromatic plant selection has risen in recent years due to increasing requirements on quality, safety, and efficiency of herbal products. Only selected varieties could guaranty the homogeneity of raw material. The chemical diversity of spontaneous populations is one of the main directions of investigations on medicinal and aromatic plants at the Institute of Botany. The target species under research are following: Origanum vulgare, Thymus spp., Hypericum spp., Achillea millefolium, and Helichrysum arenarium. The analysis of trade figures pointed out that the imported raw material mainly supplies the demands of pharmaceutical industry in Lithuania. The cultivated production covers only 4-6% of the demand for raw material. Wild plant material represents about 29% of total volume used in pharmaceutics. The trade in wild plant species and their resources is regulated by the legislations. In situ and ex situ conservation methods are used to avoid potential danger and existing threats to genetic diversity of medicinal plant species.