Total Triterpenoids Research Articles

Optimization of Ultrasonic Extraction of Triterpenes from Loquat Peel and Pulp and Determination of Antioxidant Activity and Triterpenoid Components.

The aim of this paper was to study the optimal extraction process of total triterpenes from loquat peel and pulp assisted by ultrasound. The effects of solid–liquid ratio, ethanol concentration, ultrasonic time, ultrasonic power, and ultrasonic temperature on the yield of triterpenoid acid in loquat were investigated by single-factor and response surface methodology. FRAP (Ferric ion reducing antioxidant power) method, ABTS (2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)) method, and DPPH (1,1-Diphenyl-2-picrylhydrazyl) method were used to determine the antioxidant capacity of peel and pulp at different stages. LC-MS (Liquid Chromatograph Mass Spectrometer) was used to qualitatively analyze different tissues of loquat. The optimal extraction conditions were as follows: ethanol concentration 71%, ultrasonic time 45 min, ultrasonic power 160 W, solid–liquid ratio 1:10, and ultrasonic temperature 30 °C. The total triterpenoid content of loquat peel was 13.92 ± 0.20 mg/g. The optimal extraction conditions were ethanol concentration 85%, ultrasonic time 51 min, ultrasonic power 160 W, solid–liquid ratio 1:8, and ultrasonic temperature 43 °C. The total triterpenoid content of loquat pulp was 11.69 ± 0.25 mg/g. The contents of triterpenes and antioxidant capacity in the peel and pulp of loquat at the three stages were the highest in the fruit ripening stage (S3). LC-MS analysis showed that most of the triterpenes belonged to ursolic acid derivatives and oleanolic acid derivatives, which laid the foundation for further utilization and development of loquat peel and pulp.

New Insights into Methyl Jasmonate Regulation of Triterpenoid Biosynthesis in Medicinal Fungal Species Sanghuangporusbaumii (Pilát) L.W. Zhou & Y.C. Dai.

Triterpenoids are secondary metabolites produced by the fungus Sanghuangporus baumii that have important pharmacological activities. However, the yield of triterpenoids is low and cannot meet market demand. Here, we treated S. baumii with several concentrations of MeJA (methyl jasmonate) and found that the total triterpenoid content was highest (23.31 mg/g) when the MeJA concentration was 250 μmol/L. qRT-PCR was used to quantify the transcription of five key genes involved in triterpenoid biosynthesis. The results showed that the relative transcription of most genes increased with increasing MeJA concentration, indicating that MeJA is a potent inducer of triterpenoid biosynthesis in S. baumii. To further explore whether other terpenoid biosynthesis pathways are also involved in the accumulation of triterpenoids induced by MeJA, we measured the contents of cis-Zeatin (cZ), gibberellins (GAs), and the transcript levels of related biosynthesis genes. We found that MeJA significantly inhibited the biosynthesis of cZ, GAs, and the transcription of related genes. The repressive effects of MeJA on cZ and GA accumulation were further confirmed by growth rate and biomass assays. In conclusion, our study provides an effective method to enhance the triterpenoid content of S. baumii, and also provides novel insights into the mechanism of MeJA-induced triterpenoid biosynthesis.

Absorption characteristics of ilexgenin A and ilexsaponin B1 in human umbilical vein endothelial cells after administration of the total triterpenoid saponins from Ilex pubescens.

Ilex pubescens is a famous Chinese herbal medicine, frequently used to treat cardiovascular disease in South China. In this study, we aim to explore the absorption properties of ilexgenin A (C1) and ilexsaponin B1 (C3) in vascular endothelial cells after administration of the total triterpenoid saponins from Ilex pubescens (IPTS) and clarify the possible transport mechanisms. A UPLC-qTOF-MS/MS system was used to identify the components in IPTS that could be intracellularly transported by human umbilical vein endothelial cells (HUVECs). Afterwards, a rapid, highly selective and sensitive method was established to simultaneously quantify the concentration of C1 and C3 in HUVECs after administration of IPTS. The results demonstrate that pretreatment with IPTS could promote the survival of HUVECs and reduce the damage caused by TNF-α to HUVECs. Among the main 11 components in IPTS, eight components could be absorbed by HUVECs, including seven triterpenoids and one phenolic acid. The uptake of C1 and C3 by HUVECs occurred in a time-, temperature- and concentration-dependent manner, indicating the participation of passive diffusion and active transportation mechanisms. The two triterpenoid saponins all exhibited rapid absorption and a bimodal phenomenon in their concentration-time profiles, and equilibrium could be achieved after 6h. Furthermore, C1 and C3 intracellular transportation was regulated by serum proteins, sodium-dependent glucose transporter 1 and P-glycoprotein. The current research for the first time demonstrates the in vitro pharmacokinetics characteristics of C1 and C3 in HUVECs lines, which could supply a new way of understanding the treatment of cardiovascular diseases.

병풀 부위별 기능성분 및 생리활성 분석

We investigated the antioxidant and anti-inflammatory activities in the leaf, stem, and flower of Centella asiatica (CA). CA is widely used in the medical and cosmetic industries. The leaf contained the highest total polyphenol content (236.88 mg%) and antioxidant activity when compared to the stem and flower. The total triterpenoid content was 60.65 mg/g, 54.20 mg/g, and 12.50 mg/g (leaf, flower, and stem, respectively), indicating the triterpenoids in the stem were approximately 4 to 5 times lower than the CA leaf or flower. The cell viability evaluation using RAW 264.7 and HaCaT cells revealed no significant levels of cytotoxicity (≤ 100 μg/mL). The anti-inflammatory activity assessed using RAW 264.7 cells revealed the flower had higher activity than the leaf and stem, with NO (88.3 % – inhibitory rate), TNF-α (64.8 %), IL-6 (65.7 %), and MCP-1 (28.9 %) inhibitory activity. An analysis of the inflammatory inhibition activity on the skin using HaCaT cells revealed the flower possessed a potent activity level of IL-6 (72.0 %) and IL-8 (89.4 %) inhibition. Finally, the flower and leaf had potent physiological effects and functional components. We expect that the leaf and flowers can be used as functional materials, such as antioxidant and anti-inflammatory activities.

EFFECT OF SOLVENT IN COMBINATION WITH ULTRASONIC WAVE ASSISTANCE ON TRITERPENOID EXTRACTION FROM RED LINGZHI MUSHROOM (Ganodermalucidum)

Triterpenoid is one of the compounds that contribute tored lingzhi mushroom's major significant therapeutic activities (G. lucidum). Triterpenoids have anti-allergic, anti-cancer, anti-tumor, and anti-aging effects, enhance oxygen utilization, free radical scavenging, lowerblood pressure, inhibit cholesterol biosynthesis, and enhance liver function and blood pressure, inhibit cholesterol biosynthesis, and improve liver functionetc. The beneficial effects of triterpenoids in Ganoderma lucidum on human health have generated interest in developing methods for extracting them from the fruit bodies of G. lucidum. This article presents the effect of solvent in combination with ultrasonic wave assistance in triterpenoid extraction ability from red lingzhi mushroom. Before carrying out triterpenoid extraction; Red lingzhi mushroom was ground 2mm in size. Four parameters of the extraction process, including the ethanol concentration, ratio (v:w) between ethanol solvent and mushroom, extraction time, , extraction temperature, ultrasonic time and ultrasonic intensity were determined. The extraction without ultrasonic waves for 180 minutes was used as a control. The total triterpenoid content was collected during the extraction process. The results showed that red lingzhi mushroom was extracted by ethanol solvent at a concentration of 85%, the ratio between ethanol solvent and mushroom was 3:1 (v:w), ultrasound intensity was 58 W/cm2, frequency was 20 kHz), extracting temperature was 55°C, removing time was 5 min had total triterpenoid content 3.06 times higher than the control.

WITHDRAWN: Anti-apoptotic and Anti-oxidative Effects of Aqueous Ex-tract Prepared from Steamed Daylily Flower on Normal Cardiac and Hepatic Cells against Ethanol

It is a condition of publication that manuscripts submitted to this journal have not been published and will not be simultaneously submitted or published elsewhere. Furthermore, any data, illustration, structure or table that has been published elsewhere must be reported, and copyright permission for reproduction must be obtained. Plagiarism is strictly forbidden, and by submitting the article for publication the authors agree that the publishers have the legal right to take appropriate action against the authors, if plagiarism or fabricated information is discovered. By submitting a manuscript the authors agree that the copyright of their article is transferred to the publishers if and when the article is accepted for publication.

Molecular Cloning and Analysis of an Acetyl-CoA C-acetyltransferase Gene (EkAACT) from Euphorbia kansui Liou.

Terpenoids are the largest class of natural products and are essential for cell functions in plants and their interactions with the environment. Acetyl-CoA acetyltransferase (AACT, EC2.3.1.9) can catalyze a key initiation step of the mevalonate pathway (MVA) for terpenoid biosynthesis and is modulated by many endogenous and external stimuli. Here, the function and expression regulation activities of AACT in Euphorbia kansui Liou (EkAACT) were reported. Compared with wild-type Arabidopsis, the root length, whole seedling fresh weight and growth morphology of EkAACT-overexpressing plants were slightly improved. The transcription levels of AtAACT, AtMDC, AtMK, AtHMGR, and AtHMGS in the MVA pathway and total triterpenoid accumulation increased significantly in transgenic Arabidopsis. Under NaCl and PEG treatment, EkAACT-overexpressing Arabidopsis showed a higher accumulation of total triterpenoids, higher enzyme activity of peroxidase (POD) and superoxide dismutase (SOD), increased root length and whole seedling fresh weight, and a decrease in the proline content, which indicated that plant tolerance to abiotic stress was enhanced. Thus, AACT, as the first crucial enzyme, plays a major role in the overall regulation of the MVA pathway.

The protoplast two-way fusions and fusant characteristics of Antrodia cinnamomea and Cordyceps militaris

This study generated two fused protoplasts of Antrodia cinnamomea and Cordyceps militaris in two ways. The protoplasts of A. cinnamomea were inactivated by heat to inactivate biochemical processes and enzymatic activities in the cytoplasm, and the protoplasts of C. militaris were inactivated by UV radiation to invalidate their genome function, then they were fused under optimal conditions to get a fusion rate as (7.42 ± 0.8) × 10-6 fusants/mL; the new fusants were abbreviated as Ac-Cm. On the other hand, when A. cinnamomea and C. militaris were treated with heat and UV oppositely using similar experiments, the fusion rate was (9.70 ± 0.68) × 10-5 fusants/mL, and the new fusants were abbreviated as Cm-Ac. We selected each of two best-growing fused colonies Ac-Cm-1, Ac-Cm-2, Cm-Ac-1, and Cm-Ac-2, together with parental A. cinnamomea and C. militaris, and studied their morphology, growth antagonism tests, and genetic relationships by 18S rRNA sequencing. In comparison with the initial cultures of 4 fusants, the yields of adenosine, biomass, cordycepic acid, cordycepin, total polysaccharide, and total triterpenoids were increased up 1.305−50.1563 times in the optimal medium conditions. For gene stability tests, those of the four fusants and their outputs were stabilized within 10 generations.

Cytotoxic Effects of Some Nepeta Species against Breast Cancer Cell Lines and Their Associated Phytochemical Properties.

Nepeta is one of the largest genera of the Lamiaceae family. Nepeta species are commonly employed in traditional medicine for a variety of ailments, as well as food additives. In addition, they also come to the fore with their rich phytochemical content. In the present study, the quantitative phytochemical content of methanolic extracts and infusions prepared from the aerial parts of 14 Nepeta taxa collected from Turkey and their cytotoxic effects on two breast cancer cell lines, MCF-7 and MDA-MB-231, were investigated by using the MTT (3-(4,5-dimethylthiazol-2-yl))-2,5-diphenyltetrazolium-bromide) test. According to HPLC-PDA analysis, N. racemosa methanolic extract had the highest ursolic acid content with 165.9 mg/g extract. Total sterol, total iridoid, and total triterpenoid content were determined to be greatest in the methanolic extracts of N. meyeri, N. trichocalyx and N. phyllochlamys. The MTT experiment demonstrated that certain Nepeta species suppressed the growth of MCF-7 and MDA-MB-231 cells in a dose-dependent manner. Statistical analysis revealed a strong correlation between the cytotoxic effects of the extracts and their triterpene content. In conclusion, the data obtained from this study are important in terms of forming a basis for advanced anticancer activity studies on breast cancer with Nepeta sp.

Metabolite and Gene Expression Analysis Underlying Temporal and Spatial Accumulation of Pentacyclic Triterpenoids in Jujube.

Jujube (Ziziphus jujuba Mill.) has attracted increasing attention because of its fruits’ high nutritional and medicinal value, which produce pentacyclic triterpenoids with valuable pharmacological activities beneficial to human health. However, the dynamic accumulation and metabolism pathway of triterpenoids remain unknown in jujube. Here, we performed metabolite assays of triterpenoids and expression analysis of genes involved in the corresponding metabolic processes on cultivated jujube (Z. jujuba cv. Junzao) and one type of wild jujube (Z. jujuba var. spinosa cv. Qingjiansuanzao). Our results showed that the triterpenoids accumulate predominantly in young leaves, annual stems, buds, and white-mature and beginning red stage fruit. Besides, the total triterpenoid content, ceanothic acid, oleanonic acid, and 3-ketoursolic acid were higher in ‘Qingjiansuanzao’ than in ‘Junzao’. Moreover, we found 23 genes involved in terpenoids metabolism were expressed in all organs, and the ZjSQS1, ZjCYP450/1, ZjCYP450/3, ZjOSC1, ZjFPS, and ZjAACT2 gene expression patterns were consistent with metabolites accumulation during fruit development. In addition, 100 μM MeJA induced ZjSQS1, ZjFPS, and ZjHMGR3 expression in leaves and enhanced triterpenoids accumulation. These findings will help understand the unique metabolism of terpenoids and will benefit further utilization and breeding of jujube as both edible fruit and functional food.

Phytochemical analysis, UPLC-ESI-Orbitrap-MS analysis, biological activity, and toxicity of extracts from Tripleurospermum limosum (Maxim.) Pobed

Tripleurospermum limosum (TL) has been used in folk medicine to treat gastritis. Toward the further development and use of TL, we report the phytochemical profiling, determination of active components, and antioxidant and enzyme inhibitory activities of TL. Nineteen compounds were identified by ultra-performance liquid chromatography-electrospray ionization-orbitrap-mass spectrometry for the first time in this plant. Phytochemical studies indicated that TL contained 11 types of phytochemicals. The active components [total carbohydrate content (TCC), total protein content (TProC), total triterpenoid content (TTC), total phenolic content (TPheC), total flavonoid content (TFC), total phenolic acid content (TPAC), condensed tannin content (CTC), and gallotannin content (GC)] of eight different solvent extracts were determined by ultraviolet–visible spectrophotometry. Aqueous extract had highest TProC, TPheC, and GC values. Methanol extract exhibited highest TCC and TFC values. Ethanol extract showed highest TPAC and CTC values and dichloromethane extract exhibited highest TTC value. Methanol extract showed strongest ability to scavenge 2,2-diphenyl-1-picrylhydrazyl radicals, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt cation radicals, and hydroxyl radicals, and also exhibited highest antioxidant activity in ferric-reducing antioxidant power and cupric ion-reducing antioxidant capacity assays. Best iron and copper chelating activity and H2O2 scavenging ability were shown by aqueous extract. Ethanol extract showed strongest ability to scavenge superoxide radicals and effectively prevent β-carotene bleaching. Acetone extract had highest inhibitory activity toward α-glucosidase, α-amylase, and xanthine oxidase. Ethyl ether extract had highest inhibitory activity towards urease and angiotensin converting enzyme. Aqueous and ethanol extracts had strongest inhibitory activity toward acetylcholinesterase. Methanol extract showed highest inhibitory activity toward tyrosinase. Methanol extract showed good stability and antioxidant capacity during heating, at different pH values, and after in vitro digestion and had low toxicity. The efficacy of methanol extract in stabilizing olive and sunflower oils was studied, the results suggested that methanol extract had a protective effect on the primary oxidation of the two oils. TL may be useful as a source of active components for application in human nutrition and/or phytomedicine and methanol extract of TL could be used as a natural oil stabiliser.

A simplified HPLC-UV method for the analysis of triterpenoid acids from heritage apples (Malus domestica) from western North Carolina, USA

Background: Pentacyclic triterpenoid acids are common in a number of food and spice plant species. Apples (Malus domestica) are the most common human food source for these potentially beneficial phytochemicals. Pre-20th century heritage apples have long been grown in mountainous western North Carolina and may be a wide-ranging source of these phytochemicals. Objectives: Existing extraction and analytical methods were improved and used to assess the content of three triterpenoid acids in heritage apple cultivars grown in western North Carolina, USA.Methods: Apples from a local farmers market were collected during the fall apple season. Apple peels were freeze dried, ground, and extracted with ethanol thrice. Extracts were analyzed by HPLC against external standards for betulinic, oleanolic, and ursolic acids.Results: The improved method was used to extract and to analyze the triterpenoid acid levels in 16 heritage apple cultivars grown in the Appalachian region of western North Carolina. Total triterpenoid acids ranged from 2 to 29 mg/g dry weight of peels. Content did not vary by apple color or time of harvest. Russeted varieties contained noticeably less triterpenoids.Conclusions: An improved and simplified method was used for the analysis of heritage apple varieties in western North Carolinas. A wide range of values was found for these compounds of increasing interest in the human diet and in human health.Keywords:apples, phytochemicals, triterpenoids, ursolic acid, Appalachia

Effects of Compound Elicitors on the Biosynthesis of Triterpenoids and Activity of Defense Enzymes from Inonotus hispidus (Basidiomycetes).

Inonotus hispidus has various health-promoting activities, such as anticancer effects and immune-stimulating activity. The commercialization of valuable plant triterpenoids faces major challenges, including low abundance in natural hosts and costly downstream purification procedures. In this work, orthogonal design was used to compound methyl jasmonate (MeJA), salicylic acid (SA), oleic acid, and Cu2+, and the effects of combinations on the total triterpenes biosynthesized were studied. The optimal combination was screened out and its effect on the activity of PAL, CAT, and SOD was studied. The optimal concentration of oleic acid was 2% when MeJA was 100 mol/L, and the total triterpenoid content and mycelia production were 3.918 g and 85.17 mg/g, respectively. MeJA treatment induced oxidative stress, and at the same time increased the activity of related defense enzymes. Oleic acid is thought to regulate cell permeability by recombining cell membranes. It promotes the material exchange process between cells and the environment without affecting cell growth. When oleic acid was used in combination with MeJA, a synergistic effect on triterpene production was observed. In conclusion, our findings provide a strategy for triterpenoid enrichment of I. hispidus.

An accurate, cost-effective and simple colorimetric method for the quantification of total triterpenoid and steroidal saponins from plant materials

Saponins are heterosides widely distributed in the plant kingdom. Their properties are used in many industrial sectors, such as food, cosmetics, agriculture, and pharmaceuticals, and their use is increasing due to the market trend to use natural ingredients. Although many techniques exist to quantify saponins (e.g., gravimetric, foaming, spectrophotometric or chromatographic), none of these allow simultaneous accurate, rapid and inexpensive analysis of both triterpenoid and steroidal saponins. A new colorimetric method constituted of p-anisaldehyde and sulfuric acid was developed and avoids all of the above disadvantages. Parameters used in this method allow a similar molar absorptivity for steroidal and triterpenoid saponins with high specificity in complex matrices reducing the sample preparation step and allowing quantification of saponins blends.

In vitro antioxidant activity and bioactive compounds from Calocybe indica

Nowadays, the use of mushrooms in medicine is ubiquitous and has achieved particular success. The antioxidants in mushrooms can deactivate free radicals. This study assesses the antioxidant potential of mushroom Calocybe indica with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging methods and the total antioxidant capacity. The mushroom’s ethanol extract exhibits acceptable activity with a low IC50 value (240.11 μg/mL), approximately 2.9 times lower than that of the mushroom Ophiocordyceps sobolifera extract. The ABTS scavenging rate of the extract is around 60% at 500 µg/mL, and the total antioxidant capacity is equivalent to 64.94 ± 1.03 mg of GA/g or 77.42 ± 0.42 μmol of AS/g. The total phenolics, flavonoids, polysaccharides, and triterpenoids are equivalent to 29.33 ± 0.16 mg of GAE/g, 17.84 ± 0.11 mg of QUE/g (5.04 ± 0.04%), and 4.96 ± 0.04 mg of oleanolic acid/g, respectively. Specifically, the total triterpenoid content has been reported for the first time. The mushroom can have potential biomedical applications.

Extraction of total triterpenoids from raspberry fruit and evaluation of their effects on human hepatocellular carcinoma cells

In this study, the total triterpenoids of raspberry fruit (TTRF) were prepared, and their effects on human hepatocellular carcinoma SMMC-7721 cells were investigated. SMMC-7721 cells were treated with TTRF with concentration of 0 (control), 50, 100, 200 and 400 μg/mL, respectively. The cell proliferation, cycle and apoptosis were detected. The reactive oxygen species (ROS) in cells and the expression levels of B-cell lymphoma-2 (Bcl-2) and B-cell lymphoma-2 associated X (Bax) in cells were determined. Results showed that, TTRF could obviously inhibit the proliferation of SMMC-7721 cells, arrest most cells in S and G2/M phases, and promote their apoptosis. In addition, TTRF could increase the ROS level, down-regulate the Bcl-2 protein expression and up-regulate the Bax protein expression in cells. In conclusion, cucurbitacin I has inhibitory effect on growth of SMMC-7721cells. The action mechanism may be related to its increasing ROS level and regulating of Bcl-2 and Bax expression in cells.

Plant growth, physiological variation and homological relationship of Cyclocarya species in ex situ conservation.

Natural forests of Cyclocarya paliurus have been seriously damaged because of the extreme demand for leaf medicinal uses, making conservation of this valuable, medicinal woody species necessary. Because of geographical differentiation and diverse adaptability, in this study we analysed the variations in plant growth and physiological response to environmental factors at a resource plantation of ex situ conservation and determined the homological relationships between local provenance (from Fujian Province, FJ) and introduced provenances showing high-survival rate and better growth (from Zhejiang, Hubei, Guizhou and Jiangxi Province). Our results suggested the following: (i) Plant growth: FJ had the highest plant height but not the largest basal diameter in comparison to that of other provenances. (ii) Physiological responses during the growth periods: water content in leaf of FJ had similar change with that of other provenances, except for the provenance from Guizhou Province; total soluble sugar content in leaf of FJ was more than that of other provenances; calcium content in leaf of all provenances was higher as compared to K, Mg and Na; the highest activity among four kinds of antioxidant enzymes in all provenances was superoxide dismutase, then was polyphenol oxidase and peroxidase, finally was catalase; and total flavonoid among three kinds of secondary metabolites in all provenances showed the greatest content, followed by polysaccharides and total triterpenoid. (iii) Relation analysis: plant growth and physiological responses related with environmental factors, especially temperature and precipitation. (iv) Homological relationships: leaf characteristics among six provenances varied in colour, area and common petiole length, but not the shape of leaf base or apex. Cyclocarya paliurus distributed in Fujian Province showed a very close homological relationship with that distributed in Zhejiang Province by simple sequence repeat. These findings will provide knowledge on physiological response to environmental factors and aid to select suitable provenances for Cyclocarya cultivation.

Antioxidant Properties of Triterpenoids Isolated from Bagasse-Cultivated Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), at Different Developmental Stages

Although triterpenoids are one of the main active ingredients in Ganoderma lucidum, their accumulation and antioxidant activity during the different developmental stages of G. lucidum cultivation in bagasse remains unclear. In this study, we investigated the content and antioxidant activity of total triterpenoids extracted from G. lucidum strain GL102 during the five growth stages. The obtained results showed that the highest content (12.06 mg/g) was detected in stage 3 (young fruiting body), similar to the contents of ganoderic acids B and G. However, ganoderic acids A and D exhibited maximal contents in stage 5 (spore-ejected fruiting body). The triterpenoids extracted during stage 5 were most capable of scavenging DPPH, OH, and ABTS(+) radicals, with scavenging rates of 65.88%, 86.45%, and 97.91%, respectively. Based on in vivo antioxidant assays conducted on zebrafish, the safe concentration of these triterpenoids was 0.03 mg/mL. At this concentration, the G. lucidum triterpenoids extracted during stage 5 could decrease lipopolysaccharide-induced intracellular reactive oxygen species to a level that was nearly normal (similar to the control group). The accumulation profile and antioxidant activity results reported herein provide the scientific basis needed to promote the utilization of triterpenoids derived from bagasse-cultivated G. lucidum.

Total Saponin Content (TSC) of different alfalfa (Medicago sativa L.) cultivars cultivated in field experiment

Saponins are secondary metabolites produced by various plants. These compounds have important role in the defence system. The word saponin refers to a group of different chemical compounds. Basically, sugar conjugates of triterpenoids or steroids are called saponins. Triterpene-type saponins are more specific among dicotyledonous plants, while steroid-type saponins are more characteristic of plants belonging to the monocotyledonous taxonomic group. Alfalfa is a large-scale cultivated and foraged fodder plant in Hungary. In the defence mechanism of alfalfa, saponins also play an important role. However, large amount of saponins can be toxic in animal fodder, especially in the poultry farming and piggery. As a dicotyledonous plant, the alfalfa saponins are mainly triterpenoid type. In our study we measure the total triterpenoid saponin content and leaf stem ratio of field cultivated alfalfa cultivars. Samples were collected from a randomized block design experiment, planted in the Demonstration Garden and Arboretum of Institutes of Agricultural Research and Educational Farm, in Debrecen, in 2018. Three different cultivar were investigated, and the samples were collected three different times of the growing season at growing stage of early flowering, for three years (2018-2020.). There was no difference in the TSC of examined varieties on the average of all measured data, although the sampling time showed significant effect on TSC. Our results attract attention to the fact that ageing of alfalfa stands can cause increase in TSC.

Evaluation of Nutritional Ingredients, Biologically Active Materials, and Pharmacological Activities of Stropharia rugosoannulata Grown under the Bamboo Forest and in the Greenhouse

Evaluations of the nutritional ingredients, biologically active materials, and pharmacological activities (antibacterial and antioxidant) of two types of Stropharia rugosoannulata, grown under the bamboo forest and in the greenhouse, respectively, were carried out in this study. The nutritional ingredient tests included moisture, ash, crude protein, crude fat, crude fiber, total carbohydrates, total sugar, mineral elements, vitamins, and amino acids. The biologically active material assays included total flavonoids, total polysaccharides, total triterpenoids, and reducing sugars. In addition, a broth microdilution susceptibility test was performed to evaluate antibacterial activity. The antioxidant activity was measured by ABTS+ radical scavenging, ferric-reducing power, and β-carotene bleaching assays. Results showed that two types of S. rugosoannulata were rich in protein, carbohydrate, fiber, mineral, B-group vitamins, polysaccharides, triterpenoids, and reducing sugars, with low content of fat and low Na/K ratio. Besides, they were effective in antibacterial and antioxidant activities, while S. Rugosoannulata grown under the bamboo forest was better than the greenhouse one in some respects such as higher content of protein, calcium, selenium, nicotinic acid, folic acid, polysaccharides and triterpenoids, and stronger antibacterial and antioxidant activities. They were appropriate for functional food and nutritional supplements, especially bamboo forest.