A comparative study of phytochemical metabolites and antioxidant properties of Rhodiola

Rhodiola crenulata (RC) and Rhodiola fastigiata (RF) are representative species of Rhodiola with well-accepted health benefits; the roots are the medicinal part. However, prior to this study, the differences in phytochemicals between these two species and different parts of the same species remained unclear. Using LC-ESI-MS/MS, HS-SPME-GC–MS, chemical and sensory analyses, volatile compounds and non-volatile compounds, and antioxidant activities of the roots of Rhodiola crenulata and Rhodiola fastigiata and four parts (roots, leaves, flowers, and above-ground stems) of RC were investigated. The volatile compounds and non-volatile compounds of RC roots exhibited upregulation overall compared to those of RF roots, and the odorousness, phenolic content, and antioxidant activity were more pronounced in the RC roots. The phenolic content and antioxidant activity of roots and leaves, alongside the odorousness of roots and flowers, were more significant among the four parts of RC, and the RC roots and RC flowers exhibited similar odorousness. Comparison of non-volatile differential metabolites between RC roots and RC leaves showed upregulations of saccharides and phospholipids, and minor upregulations of flavonoids and phenylpropanoids in the roots; in addition, amino acids, organic acids, and vitamins were upregulated in the leaves. These results revealed the following: 1) RC roots are superior to RF roots regarding volatile compounds and non-volatile compounds, and antioxidant activity; 2) it is more favorable to select RC roots for exploiting volatile compounds compared with RC flowers in consideration of the biomass available; 3) in terms of non-volatile compounds, and antioxidant ability, RC leaves are also of great value in addition to RC roots, though these two parts show distinct characteristics.     

Characterization of the chemical constituents in Hongjingtian injection by liquid chromatography quadrupole time‐of‐flight mass spectrometry

Hongjingtian injection is made from Rhodiola wallichiana and used in the treatment of stable angina pectoris associated with coronary heart disease. In this study, the chemical constituents in Hongjingtian injection were comprehensively studied using liquid chromatography quadrupole time‐of‐flight mass spectrometry. A total of 49 compounds were identified or assumed, including 10 organic acids, nine phenylethanoids, 10 phenylpropanoids, two flavonoid glycosides, seven monoterpene glycosides, seven octylglycosides and four other types of compounds. The structures of seven compounds were confirmed by comparing their retention times, MS and UV spectra with the corresponding authentic standards. Amongst the 49 compounds, 35 were firstly found in R. wallichiana, while they have been reported in other species of the genus Rhodiola, including Rhodiola crenulata, Rhodiola sacra, Rhodiola rosea and Rhodiola kirilowii. The possible fragmentation pathways in the mass spectrometry of the major types of compounds are proposed and summarized. Our study demonstrates a rapid method for characterizing the chemical constituents present in the Hongjingtian injection, which could also be applied to the identification of chemical constituents in other TCM formulae containing R. wallichiana.     

Brain metabolomics study for the protective effects of Rhodiola crenulata extract on Alzheimer's disease by HPLC coupled with Fourier transform‐ion cyclotron resonance mass spectrometry

In order to investigate the protective effects of Rhodiola crenulata extract on Alzheimer's disease, a brain metabolomics study in rats was conducted by high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry. Rat model was constructed by bilateral hippocampal injection of amyloid‐β peptide and immunohistochemistry was performed to evaluate the pharmacological effect of Rhodiola crenulata extract. Multivariate statistical analysis was used to discover potential biomarkers in rat brain and related metabolic pathways analysis was conducted to elucidate the action mechanism of Rhodiola crenulata extract. As a result, a total of 19 metabolites contributing to Alzheimer's disease progress were identified and nine of them were restored to the normal levels after drug administration. Pathway analysis revealed that the protective effects of Rhodiola crenulata extract are related to the regulation of glutathione metabolism and arachidonic acid metabolism in rat brain. In conclusion, this work demonstrates that the developed metabolomics method is useful to investigate the protective effects of Rhodiola crenulata extract against Alzheimer's disease. These outcomes may further provide reliable evidence to illuminate the intervention mechanism of other traditional Chinese medicines on Alzheimer's disease.     

Characterization of chemical constituents in Rhodiola Crenulate by high‐performance liquid chromatography coupled with Fourier‐transform ion cyclotron resonance mass spectrometer (HPLC‐FT‐ICR MS)

In this work, an approach using high‐performance liquid chromatography coupled with diode‐array detection and Fourier‐transform ion cyclotron resonance mass spectrometer (HPLC‐FT‐ICR MS) for the identification and profiling of chemical constituents in Rhodiola crenulata was developed for the first time. The chromatographic separation was achieved on an Inertsil ODS‐3 column (150 mm × 4.6 mm,3 µm) using a gradient elution program, and the detection was performed on a Bruker Solarix 7.0 T mass spectrometer equipped with electrospray ionization source in both positive and negative modes. Under the optimized conditions, a total of 48 chemical compounds, including 26 alcohols and their glycosides, 12 flavonoids and their glycosides, 5 flavanols and gallic acid derivatives, 4 organic acids and 1 cyanogenic glycoside were identified or tentatively characterized. The results indicated that the developed HPLC‐FT‐ICR MS method with ultra‐high sensitivity and resolution is suitable for identifying and characterizing the chemical constituents in R. crenulata. And it provides a helpful chemical basis for further research on R. crenulata. Copyright © 2016 John Wiley & Sons, Ltd.     

Metabolite profiling of sucrose effect on the metabolism of Melissa officinalis by gas chromatography-mass spectrometry

The effect of sugar on plant metabolism, which is known to be similar to hormone-like signaling, was metabolomically studied using Melissa officinalis (lemon balm). The metabolite profiles of M. officinalis treated with sucrose were analyzed by gas chromatography-mass spectrometry (GC-MS) and principal component analysis (PCA). A total of 64 metabolites from various chemical classes including alcohols, amines, amino acids, fatty acids, inorganic acids, organic acids, phosphates, and sugars were identified by GC-MS. Three groups treated with different sucrose concentrations were clearly separated by PCA of their metabolite profiles, indicating changes in the levels of many metabolites depending on the sucrose concentration. Metabolite profiling revealed that treatment with a higher sucrose level caused an increase in the levels of metabolites such as sugars, sugar alcohols, and sugar phosphates, which are related to the glycolytic pathway of M. officinalis. Furthermore, proline and succinic acid, which are associated with the proline-linked pentose phosphate pathway, the shikimic acid pathway, and the biosynthesis of phenylpropanoids, also increased with increasing sucrose concentration. Therefore, these metabolic changes induced by sucrose ultimately led to the increased production of flavonoids such as caffeic acid via the biosynthetic pathway of phenylpropanoids. This study demonstrated that the abundance changes in some primary and secondary metabolites were somewhat interlocked with each other in response to sucrose.     

Chemometric Optimization of Biologically Active Compounds Extraction from Grape Marc: Composition and Antimicrobial Activity

The article focuses on the optimization of the extraction process of biologically active compounds (BAC) from grape marc—a by-product of the wine industry. The influence of temperature, specifically 30 °C, 45 °C and 65 °C, and ethanol concentration in solutions, specifically 0–96% (v/v) on the extraction yield of polyphenols, flavonoids, tannins and anthocyanins, were investigated. The composition of individual polyphenols, anthocyanins and organic acids, antioxidant activity (DPPH and ABTS) and CIELab chromatic characteristics of the grape marc extracts (GME), were characterized. The microbiostatic and microbicidal effects in direct contact of GME with pathogenic microorganisms, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, were determined in vitro. The influence of extraction parameters on the total polyphenol content (TPC), total flavonoid content (TFC), tannin content (TC), total anthocyanin content (TAC) and their interdependencies were studied using information analysis. A mathematical model was developed on cubic spline functions. The analysis of individual compounds showed the presence of a wide range of flavonoids (procyanidin B2, procyanidin B1, hyperoside and quercetin), flavones (catechin), hydroxybenzoic acid derivatives (gallic, protocatechuic, p-hydroxybenzoic acids, m-hydroxybenzoic acid, syringic acid), hydroxycinic acid derivatives and ferulic acid methyl ester. The malvidol-3-glucoside was the main anthocyanin identified in the extract. A high amount of tartaric acid was also found. GME showed significant antimicrobial activity against Gram-positive bacteria and lower activity against Gram-negative bacteria.     

Chemical Profile,Antioxidant Properties and Antimicrobial Activities of Malaysian Heterotrigona itama Bee Bread

The aim of the study was to determine the chemical profile, antioxidant properties and antimicrobial activities of Heterotrigona itama bee bread from Malaysia. The pH, presence of phytochemicals, antioxidant properties, total phenolic content (TPC) and total flavonoid content (TFC), as well as antimicrobial activities, were assessed. Results revealed a decrease in the pH of bee bread water extract (BBW) relative to bee bread ethanolic extract (BBE) and bee bread hot water extract (BBH). Further, alkaloids, flavonoids, phenols, tannins, saponins, terpenoids, resins, glycosides and xanthoproteins were detected in BBW, BBH and BBE. Also, significant decreases in TPC, TFC, DPPH activity and FRAP were detected in BBW relative to BBH and BBE. We detected phenolic acids such as gallic acid, caffeic acid, trans-ferulic acid, trans 3-hydroxycinnamic acid and 2-hydroxycinnamic acid, and flavonoids such as quercetin, kaempferol, apigenin and mangiferin in BBE using high-performance liquid chromatography analysis. The strongest antimicrobial activity was observed in Klebsilla pneumonia (MIC₅₀ 1.914 µg/mL), followed by E. coli (MIC₅₀ 1.923 µg/mL), Shigella (MIC₅₀ 1.813 µg/mL) and Salmonella typhi (MIC₅₀ 1.617 µg/mL). Bee bread samples possess antioxidant and antimicrobial properties. Bee bread contains phenolic acids and flavonoids, and could be beneficial in the management and treatment of metabolic diseases.     

LC-MS/MS Screening,Total Phenolic,Flavonoid and Antioxidant Contents of Crude Extracts from Three Asclepiadaceae Species Growing in Jordan

This study aimed to evaluate the antioxidant activity and total phenolic content (TPC) and total flavonoid content (TFC) of crude extracts obtained from three Asclepiadaceae species, namely, Calotropis procera L., Peruglaria tomentosa L., and Pentatropis spiralis (Forsk.) Decne. Both butanol and aq. methanol extracts of the three species showed the highest amount of phenol and flavonoid contents, which exhibited the greatest antioxidant activity in the scavenging of 2,2-diphenyl-2-picrylhydrazyl free radical (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS), ferrous chelating effect (FIC), and hydroxyl radical (HDR) assays. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, sponins, flavonoids, terpenoids, and glycosides. LC-MS analysis was carried out to identify the major compounds from each crude extract. A total of 12 phenolic compounds in the extracts of the 3 species were identified and quantified, including 9 flavonoids, 2 hydroxybenzoic acids, and 3 hydroxycinnamic acids. The current study also revealed a good correlation between total phenolic contents and the observed antioxidant activity of the crude extracts.     

Phenolic Constituents of Lamium album L. subsp. album Flowers: Anatomical,Histochemical, and Phytochemical Study

Flos Lamii albi has a high biological activity and is widely used in herbal medicine. The aim of the study was to characterize the secretory structures present in Lamium album subsp. album corolla and the location of phenolic compounds. Additionally, we carried out qualitative phytochemical analyses of flavonoids and phenolic acids. Light, fluorescence, and scanning electron microscopy were used to analyze the structure of the floral organs. The main classes of phenolic compounds and their localization were determined histochemically. Phytochemical analyses were performed with high-performance thin-layer chromatography (HPTLC) and high-performance liquid chromatography (HPLC). Six types of glandular trichomes were found which contained flavonoids, phenolic acids, and tannins. The phytochemical studies demonstrated the presence of caffeic, chlorogenic, ferulic, gallic, p-coumaric, protocatechuic, syringic, gentisic, and vanillic phenolic acids as well as rutoside, isoquercetin, and quercetin flavonoids. The corolla in L. album subsp. album has antioxidant properties due to the presence of various polyphenols, as shown by the histo- and phytochemical analyses. The distribution and morphology of trichomes and the content of phenolic compounds in the corolla have taxonomic, pharmacognostic, and practical importance, facilitating the identification of the raw material.     

Chemical composition and antioxidant activity of Borago officinalis L. leaf extract growing in Algeria

The aqueous and hydroalcoholic extracts of borage (Borago officinalis) leaves from Annaba region (Algeria) were preliminary analyzed for their phenolic profile (total phenolics, total flavonoids, total flavonols, total tannins and total anthocyanins). These extracts were evaluated for their antioxidant properties by different methods such as DPPH radical scavenging, test NBT and total antioxidant activity. The two extracts have exhibited a high antiradical capacity. Indeed, the ethanolic extract showed the lower IC50 values and the highest amount of phenolics (94.09 ± 1.72 mg gallic acid/g dry extract). Using LC-MS/MS analysis, it was possible to identify phenolic acids, flavonoids, sterol and for the first time oleuropein was identified in the aqueous extract of the plant. The obtained results have demonstrated that phenolic compounds are the major contributor to the antioxidant activity of plants.     

Bioactive Compounds of Opuntia spp. Acid Fruits: Micro and Nano-Emulsified Extracts and Applications in Nutraceutical Foods

The acid fruit of the "xoconostle" cactus belongs to the genus Opuntia family of cacti. It is used as a functional food for its bioactive compounds. Several studies reported that xoconostle fruits have a high amount of ascorbic acid, betalains, phenols, tannins, and flavonoids. These compounds confer antioxidant, antibacterial, anti-inflammatory, and hepatoprotective gastroprotective activity. Xoconostle fruit extracts were tested by in vitro assays where the digestion conditions were simulated to measure their stability. At the same time, the extracts were protected by encapsulation (microencapsulation, multiple emulsions, and nanoemulsions). Applications of encapsulated extracts were probed in various food matrices (edible films, meat products, dairy, and fruit coatings). The xoconostle is a natural source of nutraceutical compounds, and the use of this fruit in the new food could help improve consumers’ health.     

Bioactive compounds,anti-inflammatory,anti-nociceptive and antioxidant potentials of ethanolic leaf fraction of Sida linifolia L. (Malvaceae)

This study investigated the bioactive compounds, anti-inflammatory, anti-nociceptive, and antioxidant properties of the ethanolic leaf fraction of Sida linifolia (ELFSL). The in vitro anti-inflammatory study employed membrane stabilization, phospholipase A2, platelet aggregation, albumin denaturation, and protease inhibition assays. Intraperitoneal injection of freshly prepared carrageenan solution (0.1 mL of 0.01 g/mL), undiluted egg albumin (0.1 mL), acetic acid (0.6 % (v/v) (10 mL/kg bw), and formalin solution (0.02 mL of 1 % v/v) into mice hind paw, were used to evaluate the anti-inflammatory and anti-nociceptive mechanisms, respectively. In vitro antioxidant potentials were determined using 1,1-Diphenyl-2-picryl-hydrazyl (DPPH), nitric oxide (NO), ferric reducing power (FRAP), and total antioxidant capacity (TAC) assays. Varying quantities of flavonoids, phenols, tannins, saponins, terpenoids, steroids, and alkaloids, were detected in the fraction. GC-FID phytochemical profiling of ELFSL revealed a high level of epicatechin, moderate levels of catechin, kaempferol, flavone, naringenin, rutin flavanones, tannins, sapogenins, proanthocyanidin, and steroids, and small amounts of sparteine, resveratrol, and lunamarine. The ELFSL exerted excellent dose-dependent in vitro anti-inflammatory activities comparable with standard drugs (aspirin/prednisolone). The LD₅₀ test showed safety up to 5000 mg/kg body weight (per oral) ELFSL. Interestingly, mice pre-administered various doses (200, 400, 600 mg/kg bw, po) of ELFSL showed significant (P < 0.05) reduction in edema, writhing, and time spent licking paw in all phases compared with control and were at par with 100 mg/kg bw (po) aspirin. The result also registered good concentration-dependent antioxidant potentials for ELFSL and was comparable to standards (gallic acid, butylated hydroxytoluene, and ascorbic acid). These imply that ELFSL possesses excellent antioxidant, anti-inflammatory, and anti-nociceptive potentials mediated by peripheral and central mechanisms.     

Acanthopanax henryi: Review of Botany,Phytochemistry and Pharmacology

Acanthopanax henryi (Oliv.) Harms (Araliaceae), also known as Eleutherococcus henryi and Caoyewujia (Hengliwujia) in Chinese, is a widely used traditional Chinese herb with the effects of expelling wind and removing dampness, relaxing the muscles and stimulating the blood circulation, and regulating the flow of qi to alleviate pain in the theory of Traditional Chinese Medicine. Acanthopanax henryi (AH, thereafter) possesses ginseng-like activities and is known as ginseng-like herb. In the past decade, a great number of phytochemical and pharmacological studies on AH have been carried out. Several kinds of chemical compositions have been reported, including terpenoids (monoterpenoids, diterpenoids, and triterpenoid saponins), phenylpropanoids, caffeoyl quinic acid derivatives, flavonoids, lignans, sterols, fatty acids, etc., among which, triterpenoid saponins were considered to be the most active components. Considerable pharmacological experiments in vitro have demonstrated that AH possessed anti-neuroinflammatory, anti-adipogenic, anti-inflammatory, antibacterial, anti-cancer, anti-oxidation, anti-AChE, anti-BuChE, and antihyaluronidase activities. The present review is an up-to-date and comprehensive analysis of the botany, phytochemistry, and pharmacology of AH.     

Phenylpropanoids from Medicinal Plants: Distribution, Classification, Structural Analysis, and Biological Activity

The literature and our own research results on phenylpropanoids from medicinal plants were reviewed and systematized. Data were presented for the distribution of phenylpropanoids. Their classification was proposed. The biological structure-activity relationships for compounds found in roseroot (Rhodiola rosea), thorny eleutherococcus (Eleutherococus senticosus), common lilac (Syringa vulgaris), purple echinacea (Echinacea purpurea), medicinal melissa (Melissa officinalis), variegated milk-thistle (Silybum marianum), and others were discussed.     

Systematically Exploring the Chemical Ingredients and Absorbed Constituents of Polygonum capitatum in Hyperuricemia Rat Plasma Using UHPLC-Q-Orbitrap HRMS

Polygonum capitatum as an ethnic medicine has been used to treat urinary tract infections, pyelonephritis and urinary calculi. In our previous study, P. capitatum was found to have anti-hyperuricemia effects. Nevertheless, the active constituents of P. capitatum for treating hyperuricemia were still unclear. In this study, an ultra-high-performance liquid chromatography coupled to quadrupole/orbitrap high-resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to comprehensively detect the chemical ingredients of P. capitatum and its absorbed constituents in the plasma of hyperuricemia rats for the first time. Xcalibur 3.0 and Compound Discoverer 2.0 software coupled to mzCloud and ChemSpider databases were utilized for qualitative analysis. A total of 114 chemical components including phenolics, flavonoids, tannins, phenylpropanoids, amino acids, amides and others were identified or tentatively characterized based on the exact mass, retention time and structural information. Compared to the previous P. capitatum study, an additional 66 different components were detected. Moreover, 68 related xenobiotics including 16 prototype components and 52 metabolites were found in the plasma of hyperuricemia rats. The metabolic pathways included ring fission, hydrolysis, decarboxylation, dehydroxylation, methylation, glucuronidation and sulfation. This work may provide important information for further investigation on the active constituents of P. capitatum and their action mechanisms for anti-hyperuricemia effects.     

Polyphenolic profile as a useful tool to identify the wood used in wine aging

Although oak wood is the main material used in cooperage, other species are being considered as possible sources of wood for the production of wines and their derived products. In this work we have compared the phenolic composition of acacia (Robinia pseudoacacia), chestnut (Castanea sativa), cherry (Prunus avium) and ash (Fraxinus excelsior and F. americana) heartwoods, by using HPLC-DAD/ESI-MS/MS (some of these data have been showed in previous paper), as well as the changes that toasting intensity at cooperage produce in each polyphenolic profile. Before toasting, each wood shows a different and specific polyphenolic profile, with both qualitative and quantitative differences among them. Toasting notably changed these profiles, in general, proportionally to toasting intensity and led to a minor differentiation among species in toasted woods, although we also found phenolic markers in toasted woods. Thus, methyl syringate, benzoic acid, methyl vanillate, p-hydroxybenzoic acid, 3,4,5-trimethylphenol and p-coumaric acid, condensed tannins of the procyanidin type, and the flavonoids naringenin, aromadendrin, isosakuranetin and taxifolin will be a good tool to identify cherry wood. In acacia wood the chemical markers will be the aldehydes gallic and β-resorcylic and two not fully identified hydroxycinnamic compounds, condensed tannins of the prorobinetin type, and when using untoasted wood, dihydrorobinetin, and in toasted acacia wood, robinetin. In untoasted ash wood, the presence of secoiridoids, phenylethanoid glycosides, or di and oligolignols will be a good tool, especially oleuropein, ligstroside and olivil, together verbascoside and isoverbascoside in F. excelsior, and oleoside in F. americana. In toasted ash wood, tyrosol, syringaresinol, cyclolovil, verbascoside and olivil, could be used to identify the botanical origin. In addition, in ash wood, seasoned and toasted, neither hydrolysable nor condensed tannins were detected. Lastly, in chestnut wood, gallic and ellagic acids and hydrolysable tannins of both the gallotannin and ellagitannin type, can be used as chemical markers.     

Secondary metabolites of Tilia tomentosa Moench inflorescences collected in Central Italy: chemotaxonomy relevance and phytochemical rationale of traditional use

Abstract

In this paper, the phytochemical analysis on the inflorescences (flowers and bracts) of a sample of Tilia tomentosa collected from an isolated population in Central Italy, was reported for the first time. Thirteen compounds (oleanolic acid, maslinic acid, apigenin, luteolin, kaempferol, quercetin, kaempferol-3-O-glucoside, quercetin-3-O-glucoside, rutin, tiliroside, ellagic acid, shikimic acid and quinic acid), belonging to several different classes of natural products, were identified on the basis of spectroscopic and spectrometric analysis. This sample was found to be mainly composed by flavonoids, followed by organic acids and pentacyclic triterpenes. The chemosystematic aspects of the identified components were also discussed, together with their pharmacologic relevance with respect to the traditional medicinal uses of this plant material.     

Terpenoids of Linaria alpina (L.) Mill. from Dolomites,Italy

This paper reports the first phytochemical analysis of Linaria alpina (L.) Mill., collected in Dolomites (Italy), a species characteristic of mountain environment. Besides aucubin (4), which is rare in the subgenus Antirrhineae of Plantaginaceae, mainly acidic compounds were found, i.e. oleanolic acid (1), ursolic acid (2) maslinic acid (3) and shikimic acid (5). The pentacyclic triterpenes of L. alpina resulted in relatively high content, whereas flavonoids resulted in low content.     

Identification of Iridoids,Flavonoids and Triterpenes from the Methanolic Extract of Melampyrum bihariense A. Kern. and the Antioxidant Activity of the Extract

Melampyrum bihariense A. Kern. (Scrophulariaceae), a plant species used in traditional medicine for the treatment of rheumatic disorders and skin infections, was investigated with regard to its antioxidant activity and identification of its bioactive chemical constituents. The crude methanolic extract of the aerial parts of M. bihariense was examined by the spectrophotometric DPPH (1,1-diphenyl-2-picrylhydrazyl) and ferric reducing antioxidant power methods. The free radical scavenging capacity (SC₅₀) of the extract was found by the DPPH method to be 27.10 mg mL^?1, and the ferric reducing ability equivalent to ascorbic acid at 50 mg mL^?1 was 0.709 μg mL^?1. The chemical composition of this highly effective in the methanolic extract was analysed, and the main compounds were isolated through solvent–solvent partition, and multiple chromatographic separations, including column chromatography, vacuum liquid chromatography, centrifugal planar chromatography and preparative thin-layer chromatography. The structures were established by one- and two-dimensional NMR and liquid chromatography-mass spectrometry. The iridoids aucubin (1), 8-epi-loganin (2) and mussaenoside (3), the flavones apigenin and luteolin and the triterpene acids ursolic acid and oleanolic acid were identified; components 2, 3, ursolic acid and oleanolic acid for the first time in this species. The present study reveals that M. bihariense exerts antioxidant activity, and the iridoids, flavonoids and triterpene acids may be the main bioactive constituents of its methanolic extract.     

Extraction and TLC Desitometric Determination of Triterpenoid Acids (Arjungenin, Arjunolic Acid) from Terminalia arjuna Stem Bark Without Interference of Tannins

The stem bark of Terminalia arjuna Linn. (fam: Combretaceae), commonly known as Arjuna in Indian systems of medicine, is a reputed drug used for various cardiac disorders. T. arjuna stem bark is reported to contain different groups of chemical constituents including phenolics, tannins, saponins and triterpenoid acids. From our earlier experience with tannin containing herbal drugs, we are aware that tannins interfere in the extraction of certain compounds and hence in their quantification. In the present experiment, we report a sample preparation method to overcome the interference of the tannins by adsorbing them with carboxy methyl cellulose, which facilitates the efficient extraction of the triterpenoid acids. Further we established TLC densitometric methods for the quantification of two of the triterpenoid acids of T. arjuna stem bark viz., arjungenin and arjunolic acid using HPTLC. The methods were validated in terms of accuracy, precision and repeatability. The calibration curve showed linearity in the range of 160–480 ng spot^?1 and 160–560 ng spot^?1 for arjungenin and arjunolic acid respectively. The percentage of arjungenin and arjunolic acid were found to be 0.324% w/w and 0.524% w/w in the stem bark by this modified method of extraction, which was many times higher than when compared to that using the extraction method without CMC (0.018% and 0.049% respectively). The study reiterates the importance of sample preparation in the quantification of non polar phytochemicals from herbal raw materials, such that the compounds of interest are extracted efficiently, overcoming the interference of other compounds like tannins in the matrix of plant material.