Total Triterpenoid Content Research Articles

Comparative studies on characterizations and cytotoxicity of oil extracted from Lingzhi (Ganoderma lucidum) G2 spore using Soxhlet extraction and microwave-assisted extraction

This study aimed to compare the characteristics, antioxidant activity, and cytotoxicity of Ganoderma lucidum G2 spore (GLS) oil extracted using two different extraction methods, including Soxhlet extraction (SLE) and microwave-assisted extraction (MAE). The SLE method yielded higher oil content than the MAE method. MAE-extracted oil showed higher lightness (L*) and yellowness (b*) values but lower redness (a*) values. Both methods provided similar fatty acid compositions in GLS oils, including palmitic acid, oleic acid, and linoleic acid as major components. Higher saturated fatty acid content was found in SLE-extracted oil, while MAE-extracted oil had increased levels of mono- and poly-unsaturated fatty acids. The oil obtained from SLE showed slightly higher di- and tri-glycerides than that obtained from MAE (P < 0.05). Furthermore, MAE-extracted oil had high oxidative stability as indicated by lower levels of conjugated diene (CD), peroxide value (PV), thiobarbituric acid-reactive substances (TBARS), and free fatty acids (FFA) as compared to SLE (P < 0.05). MAE-extracted oil had a higher total triterpenoid content than SLE-extracted oil (P < 0.05). Also, MAE-extracted oil showed lower DPPH radical scavenging activity (DPPH-RSA) than SLE-extracted oil (P < 0.05). FTIR spectra of both samples exhibited similar functional groups. Additionally, oils obtained from both processes (at 0.1–10 µg/mL) showed low toxicity on Caco-2 cells (P > 0.05). Therefore, GLS oil with high antioxidant activity and oxidative stability, especially oil extracted by MAE, could be a food supplement for health. Also, the MAE used was a potential implement for extracting oil from GLS.

Enhanced metabolite yield with compensatory biomass reduction revealed by moisture stress induction in &lt;i&gt;Centella asiatica&lt;/i&gt; (L.)

The exposure to any kind of stress tends to accelerate the secondary metabolism in medicinal plants increasing the production of secondary metabolites. The present investigation was undertaken to study the effect of moisture stress (100, 75, 50, 25 and 10% pot capacity) and control (as without plant) on growth, yield and metabolite content of Centella asiatica var. ‘Arka Prabhavi’, for two growing seasons under polyhouse conditions. Results revealed that moisture stress treatments had a significant effect on all observed growth and yield traits. Plants maintained at 100% PC exhibited luxurious vegetative growth with maximum leaf length (6.28 cm), leaf breadth (8.14 cm), petiole length (22.32 cm) and fresh biomass yield (164 g/pot). Cumulative water transpired and water use efficiency of the plants was also observed to be maximum at 100% PC. In contrary to biomass yield, increased asiaticoside (1.864%, 1.892%), madecassoside (2.856%, 3.382%) and total triterpenoid content (5.356%, 5.578%) at higher moisture stress levels of 75% and 50% PC, respectively, was observed. Hence, it is appropriate to grow Centella either at 100% or 75% PC to get optimum biomass and metabolite yield on a commercial scale.

Assessing the genetic diversity of Centella asiatica (L.) Urb. and seasonal influence on chemotypes and morphotypes in Thailand

Both genetic variation and harvesting season impact the amounts of bioactive compounds in Centella asiatica. This study investigated the seasonal influence on chemotypes and morphotypes and evaluated the genetic diversity of C. asiatica plants collected throughout Thailand. A total of 25 C. asiatica accessions was planted in the same area in Nakhon Sawan province and harvested during winter, summer, and rainy seasons. Results exhibited that variations in triterpenoid content were observed in different harvesting seasons, with total content of the four triterpenes highest during the summer season (5.98 %DW). Besides, we noted that the amount of phytochemical constituents (total phenolic, total flavonoid and total triterpenoid contents) and antioxidant were higher in winter and summer. The morphotypes were not influenced by season, while leaf area and leaf width negatively correlated with triterpenoid content. Based on chemotypes and morphtypes, we suggested the two elite accessions including Nakhon Prathom (NPT) and Nong Bua Lamphu (NBP) due to high centelloside content and large leaf. A total of 25 C. asiatica accessions was evaluated for genetic diversity by DNA fingerprints (ISSR, SCoT, and SRAP markers), resulting in a moderate genetic diversity, consisting of geography-related two populations without association with chemotypes and morphotypes. Our findings guide farmers in optimizing harvest times, and controlling centelloside content for future smart farming and insights on genetic diversity aid in future breeding and improvement of C. asiatica for medicinal industries.

Combined Metabolite and Transcriptomic Profiling Unveil a Potential Gene Network Involved in the Triterpenoid Metabolism of Rose roxburghii.

Rose roxburghii, a horticulturally significant species within the Rosa genus of the Rosaceae family, is renowned for its abundance of secondary metabolites and ascorbate, earning it the title 'king of vitamin C'. Despite this recognition, the mechanisms underlying the biosynthesis and regulation of triterpenoid compounds in R. roxburghii remain largely unresolved. In this study, we conducted high-performance liquid chromatography profiling across various organs of R. roxburghii, including fruit, root, stem, and leaves, revealing distinct distributions of triterpenoid compounds among different plant parts. Notably, the fruit exhibited the highest total triterpenoid content, followed by root and stem, with leaf containing the lowest levels, with leaf containing the lowest levels. Transcriptomic analysis unveiled preferential expression of members from the cytochrome P450 (CYP) and glycosyltransferase (UGT) families, likely contributing to the higher accumulation of both ascorbate and triterpenoid compounds in the fruits of R. roxburghii compared to other tissues of R. roxburghii. Transcriptomic analysis unveiled a potential gene network implicated in the biosynthesis of both ascorbate and triterpenoid compounds in R. roxburghii. These findings not only deepen our understanding of the metabolic pathways in this species but also have implications for the design of functional foods enriched with ascorbate and triterpenoids in R. roxburghii.

Biotransformation of bitter gourd (Momordica charantia) by Lactobacillus plantarum and its bioactivities

Bitter gourd is renowned for its various bioactivities, including antioxidant, antiglycation, and antiplatelet aggregation. However, the bioactivity and bioavailability of plant extracts could be low. Biotransformation through Lactobacillus plantarum fermentation is capable of enhancing these beneficial properties. Therefore, the present work aimed to investigate the physical and pH profiles, total phytochemical contents, bioactive contents, and bioactivities of the juices of unfermented bitter gourd (UBG) and fermented bitter gourd at 48 (FBG-48) and 96 (FBG-96) h. Fermentation gradually altered the physical and pH profile of bitter gourd, while non-significant decrease was seen in the total flavonoid content (p &gt; 0.05). The highest total phenolic content was observed in FBG-48, followed by UBG and FBG-96. UBG exhibited the highest total triterpenoid content, followed by FBG-48 and FBG-96. Interestingly, fermentation increased the antioxidant, antiglycation, and anti-aggregation activities of bitter gourd. FBG-48 demonstrated the highest antioxidant and antiglycation activities, with 10.77 and 8.68% higher activity, respectively, compared to UBG and FBG-96. Meanwhile, FBG-96 exhibited the highest antiglycation activity, with 60% increase. These could have been attributed to the biotransformation of tannic acid into syringic acid, and momordicoside Q into kuguacin P, along with the potential release of p-coumaric acid and caffeic acid from the cell walls. These findings demonstrated bitter gourd’s promising role in type-II diabetes complication treatment, particularly through its protein aggregation inhibition activity, and that fermentation could increase its bioactivities.

Proximate Composition, Phytochemical Analysis and Toxicity Assessment of Extracts of Caulerpa lentillifera using Autoclave- and Microwave-Assisted Extractions

Caulerpa lentilliferais a marine alga widely cultured and consumed in Asia and Oceania. Emerging green techniques are currently popular for phytochemical extraction. Therefore, this study aimed to investigate the proximate compositions, phytochemicals, and antioxidant properties of the C. lentilliferaextracts using autoclave-assisted and microwave-assisted extraction methods (A-CLE and M-CLE, respectively). The toxicity of extracts was tested against human normal colon cells and freshwater crustacean Daphnia magna. We found that crude lipid, ash and moisture contents in A-CLE were lower than those in M-CLE, while crude protein and crude carbohydrate were higher in A-CLE. Total flavonoid content in A-CLE (1.474 ± 0.046 mg QE g-1 extract) was higher than that in M-CLE (1.179 ± 0.054 mg QE g-1 extract). There was no difference in total phenolic contents, triterpenoid contents and antioxidant activities between the extracts. The IC50 of A-CLE (12,260 ± 197 µg mL-1) was higher than that of M-CLE (10,950 ± 169 µg mL-1). In the acute toxicity test, the LC50 of A-CLE against D. magna (7.50 ± 0.28 g L-1) was lower than LC50 of M-CLE (8.76 ± 0.26 g L-1). This study suggest that autoclave-assisted and microwave-assisted extractions are effective green methods of extracting C. lentillifera. This study will be useful for further studies of C. lentillifera extracts to improve human and animal health.

Impact of Fermentation on Hyptolide and Phytochemical Composition of Hyptis pectinata (L.) Poit

Solid State Fermentation has been employed for the biotransformation of the herbal plant Hyptis pectinata (L.) Poit is widely distributed in tropical regions of Brazil and is also present in Indonesia. The plant has been used to treat anti-inflammatory, anti-cancer, bacterial infection, and wound healing. This study aims to investigate the effects of using a solid-state fermentation method to ferment the leaves of Hyptis pectinata (L.) Poit towards the composition of phytochemicals and hyptolides. This study was carried out in several stages, including fermentation of Aspergillus niger leaves, sample extraction with methanol, determination of total phenolic and total triterpenoid content, phytochemical screening, and analysis of hyptolide using HPLC. The research results showed the growth of Aspergillus niger on the leaves of Hyptis pectinata (L.) Poit got into the logarithmic phase at 36 hours. Triterpenes and phenolics were present in the phytochemical profile, while alkaloids, tannins, and flavonoids were absent. The total phenolic and triterpenoid content increased by 33.68% and 17.78% respectively. HPLC analysis showed a decrease in hyptolide content in the fermented product. These results indicate the potential of fermented Hyptis pectinata (L.) Poit as an additional cancer therapy.

Influence of cultivation substrate on antioxidant activities and triterpenoid profiles of the fruiting body of Ganoderma lucidum.

The fruiting body of Ganoderma lucidum has been believed to possess a wide range of therapeutic effects. There are two main methods for artificial cultivation of G. lucidum to produce the fruiting body, namely wood log cultivation and substitute cultivation. The impact of cultivation substrates on the composition of bioactive compounds remains largely unexplored. This study aims to compare the antioxidant activities and triterpenoid profiles of the fruiting bodies of G. lucidum that cultivated through wood log cultivation (WGL) and substitute cultivation (SGL) methods. The antioxidant activities, including the DPPH radical scavenging, hydroxyl radical scavenging, superoxide radical scavenging, and total antioxidant activities, were assessed in both WGL and SGL samples. Furthermore, the UHPLC-Q-Orbitrap-MS technique was employed to compare their phytochemical profiles, with a specific emphasis on triterpenoid constituents. It was found that WGL samples exhibited significantly higher total triterpenoid content, DPPH radical scavenging activity, and total antioxidant activity. Furthermore, an untargeted metabolomics approach employing UHPLC-Q-Orbitrap-MS tentatively identified a total of 96 triterpenoids. Distinguishingly different triterpenoid profiles between the two types of G. lucidum samples were revealed via the utilization of principal component analysis (PCA) and hierarchical cluster analysis (HCA). Specifically, 17 triterpenoids showed significant differences. Of these triterpenoids, 6 compounds, such as ganosporelactone B, ganoderol A, ganoderic acid A, ganoderic acid alpha, were significantly higher in SGL samples; 11 compounds, such as lucidenic acid A, lucidenic acid D1, lucidenic acid F, lucidenic acid G, lucidenic acid J, ganoderic acid E, and ganoderic acid O, were significantly higher in WGL samples. These findings expand our knowledge regarding the impact of cultivation substrate on the antioxidant activities and triterpenoid profiles of G. lucidum, and offer practical implications for its cultivation.

A Sustainable Extraction Approach of Phytochemicals from Date (Phoenix dactylifera L.) Fruit Cultivars Using Ultrasound-Assisted Deep Eutectic Solvent: A Comprehensive Study on Bioactivity and Phenolic Variability.

The present study aimed to evaluate the efficacy of natural deep eutectic solvents (NADESs) on the extraction of phytochemicals from eight Algerian date fruit cultivars (Phoenix dactylifera L.). In this study, lactic acid/sucrose-based NADESs were used as an alternative to conventional chemical solvents using the ultrasound-assisted extraction (UAE) method. The obtained extracts were assessed for the determination of bioactive compound contents, phenolic composition, antioxidant activity, and enzyme inhibitory potential. The results showed a considerable variation in phytochemical compositions and related activities between cultivars, where the greatest contents of total phenolics (1288.7 mg GAE/100 g), total flavonoids (53.8 mg QE/100 g), proanthocyanidins (179.5 mg CE/g), and total triterpenoids (12.88 mg OAE/100 g) were detected in the fruits of the Ourous cultivar. The same cultivar displayed the highest antioxidant capacity against DPPH• free radical (595 mg AAE/100 g), ABTS•+ cation radical (839 mg TE/100 g), and ferric reducing antioxidant potential (704 mg AAE/100 g). All extracts manifested moderate antioxidant activities tested by phosphomolybdenum, NO•, and linoleic acid lipid peroxidation assays. These extracts also exhibited interesting levels of in vitro enzyme inhibition; the Ourous cultivar gave the best inhibitory activity against α-amylase and acetylcholinesterase with 45 and 37%, respectively. HPLC-DAD-MS detected a total of five compounds, with phenolic acids and flavonoids being the main phenolics identified in the extract. The phenolic composition exhibited significant variability among cultivars. Notably, the highest amounts were revealed in the Tazizaout cultivar, with the predominance of gallic acid. The results confirmed that the combination of UAE and NADESs provides a novel and important alternative to chemical solvents for sustainable and environmentally friendly extraction and can represent a good alternative in food and pharmaceutical industry applications.

Harnessing postharvest light emitting diode (LED) technology of Centella asiatica (L.) Urb. to improve centelloside content by up-regulating gene expressions in the triterpenoid pathway

Centella asiatica (L.) Urb. has wound healing, anti-inflammatory, cognitive improvement, and neuroprotective properties which have been attributed to its centelloside content. However, the quantities of these bioactive compounds are limited and vary due to genetic and environmental factors. Light qualities are known to enhance the production of secondary metabolites in several plant species, both preharvest and postharvest. In this study, fresh leaves of C. asiatica were subjected to different light emitting diode (LED) quality including white, dark, red, blue, and green to assess centelloside content, phytochemical constituents, and transcription level expression of triterpenoid biosynthesis genes. Results showed that white and blue LEDs significantly increased centelloside content in C. asiatica leaves at 3 days postharvest (dph) by 73 % over the control group at 0 dph. Blue LEDs stimulated the expression of triterpenoid biosynthesis genes including C. asiatica squalene synthase (CaSQS), C. asiatica β-amyrin synthase (CabAS), and C. asiatica UDP gluclosyltransferase-73AH1 (CaUGT73AH1; CaUGT), while different LED conditions gave diverse results. Red LED treatment triggered higher total flavonoid content (TFC) and total triterpenoid content (TTC) while white LEDs enhanced total triterpenoid content (TTC). Taken together, our findings suggest that postharvest under blue LEDs is a great approach to increase centelloside production of C. asiatica through gene up-regulation in triterpenoid pathway. Therefore, postharvest technology by LEDs serves as an effective tool for improving raw material quality for medicinal plant industries.

Exogenous Application of Coconut Water to Promote Growth and Increase the Yield, Bioactive Compounds, and Antioxidant Activity for Hericium erinaceus Cultivation

Hericium erinaceus (Bull.:Fr) Pers. is a medicinal mushroom that has various health benefits and is a rich source of bioactive compounds and antioxidant activity. In recent years, H. erinaceus has been considered for its many medicinal properties and is widely consumed in Asian countries. This work aimed to explore the potential improvement of coconut water utilization in order to promote growth and increase the yield, as well as the enhancement of bioactive compounds and antioxidant activity of H. erinaceus. The application of coconut water at a concentration of 20% (v/v) resulted in the optimal development and production of H. erinaceus. In terms of biological efficiency, it was demonstrated that the 22.09% increase over the control treatment resulted in a higher yield. Moreover, H. erinaceus cultivated from coconut water intake at all concentrations resulted in higher protein content. Additionally, bioactive compounds such as total triterpenoid content and total phenolic content of H. erinaceus were 67.87–89.24 mg Urs/g DW and 16.62–17.39 mg GAE/g DW, respectively, while H. erinaceus grown on a control treatment had the lowest at 56.84 mg Urs/g DW and 14.42 mg GAE/g DW, respectively. Applying coconut water at all concentrations resulted in H. erinaceus IC50 values of 0.58–0.69 mg/mL exhibiting higher DPPH activities than those grown on control treatment IC50 value of 0.77 mg/mL. Therefore, this finding of the study indicated that the utilization of coconut water as a therapy resulted in a significant enhancement in the development, production, and bioactive compounds of H. erinaceus, in comparison to the control treatment.

Phytochemical Constituent and Antioxidant Activity Evaluation of Red Seaweed Eucheuma sp.

Oxidative stress is a condition in which there is an imbalance between production of free radicals and protective response via antioxidant system. There are endogenous and exogenous antioxidants, however as age increases, there is a reduction in endogenous antioxidant, thus the search for potential exogenous antioxidants which could be derived from natural resources are needed. Indonesia is a megabiodiversity country which has more than 30,000 species of plants and animals. Red seaweed Eucheuma sp. is one of marine macroalgae species which shows potent biological activities. This study aims to determine the phytochemical constituent and to evaluate the antioxidant activity of red algae Eucheuma sp. Seaweed Eucheuma sp. obtained from Lombok, Nusa Tenggara Barat, Indonesia, were extracted by maceration process using three solvents, n-hexane, ethyl acetate, and ethanol, sequentially. Each extract was analyzed for its phytochemical constituents by phytochemistry screening, thin layer chromatography, total phenolic content, total flavonoid content, and total triterpenoid content. Evaluation of antioxidant activity for ethyl acetate extract and ethanol extract were done using DPPH method. Phytochemical analysis of Eucheuma sp. shows positive result for steroid and triterpenoid. Thin layer chromatography analysis of the Eucheuma sp. extracts showed total of 11 phytochemical constituents. Quantitative analysis revealed that the highest value in ethyl acetate extract, with total phenolic content of 29.57 mg gallic acid equivalent/g extract, total flavonoid content of 0.54 mg quercetin equivalent/g extract, and total triterpenoid content of 1.08 mg ursolic acid equivalent/g extract. Moreover, ethylacetate extract of Eucheuma sp. demonstrated an active antioxidant activity against DPPH free radical with IC50 value of 27.96 µg/mL. Thus, ethylacetate extract of Eucheuma sp. derived from Lombok, Indonesia, has a potential to be developed as a natural antioxidant.

Effects on the Synthesis and Accumulation of Triterpenes in Leaves of Cyclocarya paliurus under MeJA Treatment

Cyclocarya paliurus (Batal.) lljinskaja, the sole and multi-functional tree species of the family Juglandaceae, grows extensively in subtropical areas of China. Species-specific triterpenoids in its leaves have validated health-promoting effects, including hypoglycemic and hypolipidemic activities. To illustrate the effect of MeJA treatment on the accumulation and biosynthesis of triterpenoids in C. paliurus leaves at different stages of maturity, the contents of total triterpenoids and six triterpene compounds, along with the relative expression of three key genes, were detected. The results showed that the contents of triterpenes and expression patterns of the genes significantly differed among the samples. Different treatment times also had diverse effects on triterpenoid accumulation and gene expression. MeJA treatment had positive effects on total triterpenoids, cyclocaric acid B, and cyclocarioside A/B, especially in young leaves. Gene expression was highest in young leaves after 10 days of treatment, indicating that they were the most sensitive to MeJA. This study provided a reference for improving the accumulation of triterpenoids in C. paliurus plantations in the future.

Evaluation of bioactive compounds, antioxidant capacity and mineral elements in the leaves of guava (Psidium guajava L.) genotypes from China

Guava (Psidium guajava L.) is a medicinal tree species and its leaves are rich in beneficial bioactive compounds with antioxidant capacity. China has the second-largest planting area in the world however a comprehensive evaluation of the antioxidant capacity of guava genotypes in China has not yet been conducted yet. To identify the desirable genotypes with the greatest antioxidant capacity, 18 guava genotypes widely cultivated in China were collected in a germplasm garden. In this study, the antioxidant capacity, bioactive compound contents and mineral element contents of guava leaves were assessed. Genotypic variation was observed for antioxidant capacity, bioactive compound contents and mineral element contents in guava leaves. The bioactive compound contents were all significantly and positively correlated with the antioxidant capacity in guava leaves. Correlation analysis indicated that the total phenolic total triterpenoid contents were significantly related to the total phosphorus content. Principal component analysis and cluster analysis showed that DW was superior in antioxidant capacity (radical scavenging abilities of DPPH and ABTS) and total phenolic, total flavonoid and total triterpenoid contents, and the medicinal quality of superior guava genotypes was regulated by the leaf mineral element contents, especially phosphorus. This study is not only helpful for developing guava genotypes with superior medicinal quality in the future but can also provide theoretical support for fertilization management in the cultivation of superior guava genotypes to achieve higher medicinal quality.

Plant species richness regulated by geographical variation down-regulates triterpenoid compounds production and antioxidant activities in white birch bark

White birch (Betula platyphylla Suk.) is a pioneer tree species widely distributed in the high northern hemisphere, and its bioactive compounds have ecological and traditional medicinal roles. How ecological factors impact triterpenoid production is not well understood yet. This study examined the effects of geo-climate, soil properties and plant community properties on triterpenoid compounds synthesis and antioxidant activities of white birch bark at eight locations (13° latitude and 6° longitude ranges) in Northeast China. The results showed that the contents of total triterpenoids, betulin and betulinic acid in white birch bark were high at higher latitudes, and low at lower latitudes. The antioxidant activity of the extracts analyzed by DPPH (1, 1-diphenyl-2-picrylhydrazyl), ABTS (2, 2-azinobis-3-ethylbenzothiazoline-6-sulphonic acid), and FRAP (ferric reducing antioxidant power assay) methods varied across different locations and positively correlated with triterpenoid compound contents. Redundancy analysis indicated that more than 40% of the secondary metabolite variations could be explained by plant species richness in the community, largely regulated by geographical variation. Our findings highlighted that higher species richness reduced triterpenoid accumulation and antioxidant potential. At the same time, the harsh environments in cold and dry geo-climate increased triterpenoid production of white birch. This study favors the efficient use of white birch for triterpenoid production and implies that the stress of harsh environment will promote the synthesis of the secondary metabolite synthesis of dominant trees.

Development of a Transformation System for the Medicinal Fungus Sanghuangporus baumii and Acquisition of High-Value Strain

To further explore the molecular mechanism of triterpenoid biosynthesis and acquire high-value strain of Sanghuangporus baumii, the Agrobacterium tumefaciens-mediated transformation (ATMT) system was studied. The key triterpenoid biosynthesis-associated gene isopentenyl diphosphate isomerase (IDI) was transformed into S. baumii by ATMT system. Then, the qRT-PCR technique was used to analyze gene transcript level, and the widely targeted metabolomics was used to investigate individual triterpenoid content. Total triterpenoid content and anti-oxidant activity were determined by spectrophotometer. In this study, we for the first time established an efficient ATMT system and transferred the IDI gene into S. baumii. Relative to the wild-type (WT) strain, the IDI-transformant (IT) strain showed significantly higher transcript levels of IDI and total triterpenoid content. We then investigated individual triterpenoids in S. baumii, which led to the identification of 10 distinct triterpenoids. The contents of individual triterpenoids produced by the IT2 strain were 1.76–10.03 times higher than those produced by the WT strain. The triterpenoid production showed a significant positive correlation with the IDI gene expression. Besides, IT2 strain showed better anti-oxidant activity. The findings provide valuable information about the biosynthetic pathway of triterpenoids and provide a strategy for cultivating high-value S. baumii strains.

Squalene epoxidase (SE) gene related to triterpenoid biosynthesis assists to select elite genotypes in medicinal plant: Cyclocarya paliurus (Batal.) Iljinskaja

Triterpenoids, known for their anti-inflammatory, anticancer, and hypoglycemic properties, are the major bioactive components in Cyclocarya paliurus (Batal.) Iljinskaja. Selecting elite individuals with high triterpenoids content is the basis of C. paliurus industry for medicinal use. In this study, seasonal variation patterns of total triterpenoids and five triterpene monomers accumulation for three groups with different total triterpenoid contents (TTC; H: 59.74–64.03 mg g−1; M: 47.66–57.08 mg g−1, and L: 35.26–42.22 mg g−1) were surveyed. Seasonal expression dynamics of 6 key genes relevant to triterpenoids biosynthesis, including HMGR, DXR, SQS, SE, LUS, and β-AS, were described by quantitative real-time PCR (qRT-PCR) for three groups. The expression levels of HMGR, SE, LUS, and β-AS genes in group H were higher than in groups M and L. In addition, Pearson correlation analysis showed that they were significantly positively correlated with triterpene accumulation, and the expression level of SE gene not only was significantly correlated with downstream genes, but also exhibited a linear relationship with TTC, especially in September. These results suggest that SE gene could serve as an effective make for screening elite individuals with high TTC from the germplasm of C. paliurus for medicinal use. Further testing on randomly selected individuals in next September proved the feasibility and reliability of SE gene in assisted selection. Also, we successfully cloned the full-length cDNA of SE. Thus, our work provides an efficient way to attain superior genotypes to develop medicinal industry of C. paliurus in practice.

Triterpenoids of Three Apple Cultivars-Biosynthesis, Antioxidative and Anti-Inflammatory Properties, and Fate during Processing.

Triterpenoids are a group of secondary plant metabolites, with a remarkable pharmacological potential, occurring in the cuticular waxes of the aerial parts of plants. The aim of this study was to analyze triterpenoid variability in the fruits and leaves of three apple cultivars during the growing season and gain new insights into their health-promoting properties and fate during juice and purée production. The identification and quantification of the compounds of interest were conducted using gas chromatography coupled with mass spectrometry. The waxes of both matrices contained similar analytes; however, their quantitative patterns varied: triterpenic acids prevailed in the fruits, while higher contents of steroids and esterified forms were observed in the leaves. The total triterpenoid content per unit area was stable during the growing season; the percentage of esters increased in the later phases of growth. Antioxidative and anti-inflammatory properties were evaluated with a series of in vitro assays. Triterpenoids were found to be the main anti-inflammatory compounds in the apples, while their impact on antioxidant capacity was minor. The apples were processed on a lab scale to obtain juices and purées. The apple purée and cloudy juice contained only some of the triterpenoids present in the raw fruit, while the clear juices were virtually free of those lipophilic compounds.

Protective Effect of Hawthorn Fruit Extract against High Fructose-Induced Oxidative Stress and Endoplasmic Reticulum Stress in Pancreatic β-Cells

Hyperglycemia has deleterious effects on pancreatic β-cells, causing dysfunction and insulin resistance that lead to diabetes mellitus (DM). The possible causes of injury can be caused by glucose- or fructose-induced oxidative and endoplasmic reticulum (ER) stress. Hawthorn (Crataegus pinnatifida) fruit has been widely used as a hypolipidemic agent in traditional herbal medicine. The study aimed to investigate whether high fructose-induced pancreatic β-cell dysfunction could be reversed through amelioration of ER stress by the treatment of polyphenol-enriched extract (PEHE) from hawthorn fruit. The extract was partitioned using ethyl acetate as a solvent from crude water extract (WE) of hawthorn fruits, followed by column fractionation. The results showed that the contents of total polyphenols, flavonoids and triterpenoids in PEHE could be enhanced by 2.2-, 7.7- and 1.1-fold, respectively, in comparison to the original obtained WE from hawthorn fruit. In ER stress studies, a sharp increase in the inhibitory activity on the gene expression levels of GRP79, ATF6, IRE1α and CHOP involved in ER stress was evident when dosages of PEHE at 50-100 μg/mL were used against high-fructose (150 mM)-treated cells. HPLC-MS/MS analysis showed that polyphenols and flavonoids collectively accounted for 87.03% of the total content of PEHE.

Persuasive and Strategic Approaches for Preliminary Phytochemical Screening, Quantitative Analysis and Antioxidant Activity of Crude Plant Extracts from Clematis napaulensis DC Indigenous to Nagaland, India

The present study is designed to extract and investigate the bioactive phytoconstituents, their secondary metabolites, and antioxidant potential from the leaf extracts of Clematis napaulensis belonging to the family Ranunculaceae. Extract was investigated for its preliminary phytoconstituents by chemical analysis method. Quantification of total phenolic content by Folin-Ciocalteau assay; Total Flavonoid Content and Total Triterpenoid Content (TTC) were quantified by colorimetric method. Antioxidant potential was evaluated in vitro using Phosphomolybdenum assay, Hydroxyl radical scavenging activity, Reducing power assay, and the DPPH method. The preliminary phytochemical screening results suggested that a wide variety of pharmacologically active phytoconstituents such as phenolic compounds, alkaloids, glycosides, flavonoids, saponins, tannins, triterpenoids, gums, proteins, and reducing sugars were present whereas non-reducing sugar was absent. The total phenolic content in the methanolic extract was 0.8308 μg/mL of gallic acid equivalent; the total flavonoid content of the plant was 1.3850 μg/mL of quercetin equivalent and the total triterpenoid content was found to be 1.414 μg/mL of oleanolic acid equivalent. In vitro antioxidant studies revealed that Clematis napaulensis possess good antioxidant potential. This is the first report of Clematis napaulensis, the phytochemical screening results suggested that the extract would be a potential alternative also it can be explored to discover and development of new chemical moieties for treating various diseases.