Ethyl Acetate Portion Research Articles

Structural Elucidation and Complete NMR Spectral Assignments of Monascus Monacolin Analogs.

One new monacolin analog, monacolin V (1), together with two new monacolin-like natural products, 6-hydroxyl monacolin P (2) and 3-keto monacolin S (3), were isolated from the ethyl acetate portion of red yeast rice ethanol extract. Their structures were identified by HRESIMS and NMR experiments, and the complete assignments of 1H and 13C NMR data for three compounds were obtained by the aid of HSQC, HMBC, 1H-1H COSY, and NOESY data. This is the first time that the NMR data of compounds 2 and 3 have been fully assigned.

Structural elucidation and complete NMR spectral assignments of Monascus monacolin analogues.

Three new monacolin analogues, 3,6-dihydroxy-monacolin P (1), 6-methoxy monacolin S (2), and 6-methoxy dehydromonacolin S (3), were isolated from a fraction that strongly inhibited 3-hydroxy-3-methylglutaryl-CoA reductase from the ethyl acetate portion of red yeast rice ethanol extract. Their structures were determined through a combination of 1D and 2D NMR experiments, mass spectrometry analysis, and known literature reports.

LC-MS/MS method for quantitative profiling of ketone bodies, α-keto acids, lactate, pyruvate and their stable isotopically labelled tracers in human plasma: An analytical panel for clinical metabolic kinetics and interactions

An important area within clinical research is in vivo metabolism of ketone bodies (β-hydroxybutyrate and acetoacetate) and in connection metabolites that may affect their production and/or cellular transport such as the keto-acids from the branched-chain amino acids, lactate and pyruvate. To determine in vivo metabolite turnover, availability of accurate and sensitive methods for analyzing the plasma concentrations of these metabolites and their stable isotopically labeled enrichments is mandatory. Therefore, the present study describes a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for simultaneous analysis of ketone bodies, α-keto acids, lactate, pyruvate, and their tracer enrichments in humans using 2 different derivatization techniques with 4-bromo-N-methylbenzylamine and O-benzylhydroxylamine as derivatization reagents, and 1-ethyl-3-dimethylaminopropyl carbodiimide as coupling compound followed by a single LC-MS/MS run. The method was validated for matrix effects, linearity, accuracy, precision, recovery, stability, and enrichment (ratio) analysis of a stable isotopically labelled analytes (tracers) continuously infused in humans divided by the unlabeled endogenous analyte (tracee) that makes it possible to quantify the analyte in vivo synthesis and degradation rates. The applied parallel derivatization procedure yielded good sensitivity for all analytes of interest and their tracers. Despite the double derivatization method, mixing the ethyl acetate portions at the final stage made it possible to simultaneously analyze all compounds in a single LC-MS/MS run. Moreover, the liquid chromatography method was optimized to robustly quantify the keto acids derived from leucine (α-keto-isocaproic acid) and isoleucine (α-keto-β-methylvaleric acid), the compounds with similar chemical structure and identical molecular weights. The presented method is designed and validated for human plasma. However, care should be taken in blood sampling and processing procedures as well as quick freezing and storage at −80 °C due to the instability of especially acetoacetate.

Chromatography fractions from the ethyl acetate fraction of Harungana madagascariensis Lam. Ex Poiret fruits (Hypericaceae) inhibited the growth of Mycobacterium tuberculosis

The economic burden of drug resistant tuberculosis strain is enormous due to its being one of the leading cause of global mortality. New treatments that are faster, simpler and affordable are urgently needed. Thus this research aimed to validate that the ethyl acetate fraction from the Nigeria variety of Harungana madagascariensis contain secondary metabolites with anti-Mycobacterium tuberculosis activity. The fruit of H. madagascariensis was extracted with 70% aqueous ethanol by cold maceration. The crude aqueous ethanol extract was defatted with n-hexane by partitioning and the aqueous portion further partitioned with ethyl acetate to give ethyl acetate portion (EAP) used in this study. Fractionation of EAP was done using chromatographic techniques. Phytochemical screening of the fractions was done using standard methods. Anti- Mycobacterium tuberculosis screening was done using Lowestein Jensen(LJ) method (test fractions stock solution 1mg/ml diluted to a 0.01mg/ml in the LJ medium while isoniazid, dihydrostreptomycin, ethambutol and rifampicin at their reported minimum inhibition concentrations(MIC) were used as reference anti-TB drugs). Four (4) chromatography fractions (EAF1 –EAF4) were obtained. All fractions were found to contain phenolics with anthraquinone present in EAF2 and EAF3. Fractions EAF3 and EAF4 inhibited the growth of the Mycobacteria tuberculosis. This preliminary screening validates the use of H. madagascariensis in traditional medicine and as a potential source of natural products with anti-Mycobacterium tuberculosis activity as well as a good source of anthraquinone.

Stilbenoids and Flavonoids from Cajanus cajan (L.) Millsp. and Their α-Glucosidase Inhibitory Activities.

Two new stilbenoids, cajanstilbenoid C (1) and cajanstilbenoid D (2), together with eight other known stilbenoids (3-10) and seventeen known flavonoids (11-27), were isolated from the petroleum ether and ethyl acetate portions of the 95% ethanol extract of leaves of Cajanus cajan (L.) Millsp. The planar structures of the new compounds were elucidated by NMR and high-resolution mass spectrometry, and their absolute configurations were determined by comparison of their experimental and calculated electronic circular dichroism (ECD) values. All the compounds were assayed for their inhibitory activities against yeast α-glucosidase. The results demonstrated that compounds 3, 8-9, 11, 13, 19-21, and 24-26 had strong inhibitory activities against α-glucosidase, with compound 11 (IC50 = 0.87 ± 0.05 μM) exhibiting the strongest activity. The structure-activity relationships were preliminarily summarized. Moreover, enzyme kinetics showed that compound 8 was a noncompetitive inhibitor, compounds 11, 24-26 were anticompetitive, and compounds 9 and 13 were mixed-competitive.

ISOLATION OF STIGMASTEROL AND GASTROPROTECTIVE ACTIVITY OF THE ETHYL ACETATE PORTION FROM ACACIA NILOTICA LINN (FABACEAE) SEEDS

Acacia nilotica Linn is used traditionally in the treatment of various diseases, such as diarrhea, ulcer, dysentery, asthma, inflammation and cancer. The present study aimed at isolation and characterization of a steroidal compound and evaluating the gastroprotective activity of the ethyl acetate portion of A. nilotica seeds in indomethacin-induced ulcer model. Fractionation of the ethyl acetate portion using standard chromatographic techniques led to the isolation of a compound (B2-I) which on further purification and characterisation using physical, chemical and spectral analysis, and by comparison with literature values was identified as stigmasterol. The effect of ethyl acetate portion of A. nilotica on experimentally induced ulcer was dose-dependent with curative ratios of 40%, 60% and 75% at concentrations of 100, 200 and 300 mg/kg body weights respectively. It significantly (P<0.05) decreased free and total acidity and increased the pH of gastric juice with respect to the indomethacin treated group. This justifies the ethnomedicinal use of A. nilotica seed as an antiulcer agent. The study recommends that A. nilotica seeds be evaluated against other ulcer models such as ethanol and stress induced ulcers.

New chromones from the roots of Saposhnikovia divaricata (Turcz.) Schischk with anti-inflammatory activity

Fifteen new chromones, sadivamones A-E (1–5), cimifugin monoacetate (6), sadivamones F-N (7–15), together with fifteen known chromones (16–30), were isolated from the ethyl acetate portions of 70% ethanol extract of Saposhnikovia divaricata (Turcz.) Schischk roots. The structures of the isolates were determined using 1D/2D NMR data and electron circular dichroism (ECD) calculations. Meanwhile, LPS induced RAW264.7 inflammatory cell model was used to determine the potential anti-inflammatory activity of all the isolated compounds in vitro. The results showed that compounds 2, 8, 12–13, 18, 20–22, 24, and 27 significantly inhibited the production of lipopolysaccharide (LPS)-induced NO in macrophages. To determine the signaling pathways involved in the suppression of NO production by compounds 8, 12 and 13, we investigated ERK and c-Jun N-terminal protein kinase (JNK) expression by western blot analysis. Further mechanistic studies demonstrated that compounds 12 and 13 inhibited the phosphorylation of ERK and the activation of ERK and JNK signaling in RAW264.7 cells via MAPK signaling pathways. Taken together, compounds 12 and 13 may be valuable candidates for the treatment of inflammatory diseases.

Two New Phenolic Glycosides with Lactone Structural Units from Leaves of Ardisia crenata Sims with Antibacterial and Anti-Inflammatory Activities.

Two new lactones, named Ardisicreolides A–B (1–2), together with four known flavonoids, Quercetin (3), Myricetrin (4), Quercitrin (5), Tamarixetin 3-O-rhamnoside (6), were isolated from the ethyl acetate portion of 70% ethanol extracts of dried leaves from Ardisia crenata Sims. These compounds were identified from Ardisia crenata Sims for the first time. The structures of 1–6 were elucidated according to 1D and 2D-NMR methods and together with the published literature. All of the isolated compounds were evaluated for in vitro anti-microbial effect against Escherichia coli, Pseudomonas aeuroginosa, Enterococcus faecalis, Proteus vulgaris, Staphylococcus aureus, and Bacillus subtilis. In addition, compounds 1–2 were assessed for anti-inflammatory activity by acting on LPS-induced RAW 264.7 macrophage cells in vitro. The results showed that only compound 2 exhibited moderate antibacterial activity on Bacillus subtilis. Moreover, compounds 1 and 2 were found to significantly inhibit the production of nitric oxide (NO) and reduce the release of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4), and interleukin-10 (IL-10) in LPS-induced RAW 264.7 macrophage cells. The present data suggest that lactones from the leaves of A. crenata Sims might be used as a potential source of natural anti-inflammatory agents.

A new hemiacetal chromone racemate and α-glucosidase inhibitors from Ficus tikoua Bur

From the petroleum ether and ethyl acetate portions of the 95% ethanol extract of Ficus tikoua Bur., a new hemiacetal chromone racemate, named (±)-ficunomone (1), together with twenty-two known flavonoids (2-23) were isolated. The new structure was elucidated by NMR, HRESIMS, and X-ray single-crystal diffraction analysis, and the known structures were determined by comparison of spectroscopic data with those reported from literatures. All the compounds were assayed for their inhibitory activities against yeast α-glucosidase, seven flavonoids could inhibit α-glucosidase, among which compounds 22 and 23 exhibited the highest inhibitory activity, with IC50 values at 5.12 ± 0.10 and 3.43 ± 0.15 μM respectively. Preliminary structure and relationship activity of all the compounds was analysed. Kinetic analysis of compounds 22 and 23 indicated that they are both uncompetitive inhibitors. Molecular docking studies revealed that they bound to amino acid residues of the α-glucosidase activity pocket.

Isolation of bioactive constituents from the ethylacetate leaf extract portion of Gmelina arborea (Verbenaceae)

The medicinal value of plants has assumed important dimension in the past few decades owing mainly to the discovery that extracts from plants contain not only minerals and primary metabolites but also a diverse array of secondary metabolites with anti-inflammatory, antioxidant and antimicrobial potential. In Nigeria (West Africa), the decoction of the leaf extract of Gmelina arborea mixed with trona is taken as a remedy for Malaria fever, Typhoid as well as an antibacterial agent. The main objective of the present research was to carry out preliminary phytochemical and antimicrobial evaluation of the ethyl acetate portion of the leaf extract of Gmelina arobrea using standard protocols. Preliminary phytochemical screening revealed the presence of Alkaloids, Flavonoids, Saponins, Phenolic compounds, Triterpenes and Steroids. Compound 1 isolated was determined as 2-(2,4- dihydroxy-5-prenylphenyl)-5,6- methylenedioxybenzofuran, while compound 2 was determined as 5,7,2',4'-tetrahydroxy-8-lavandulyl-5'- methoxy flavone. Determination of the antimicrobial activity of the ethyl acetate portion on some isolates was conducted. The zone of inhibition from the antibacterial activity from the ethyl acetate portion was found to be fairly good against gram (+) bacteria e.g Staphlococcus aureus, Bacillus subtilis and gram (-) bacteria e.g Klebsiela pneumonia and Escherichia coli. The antifungal activity of the ethyl acetate portion was found to be active against Aspergillus niger, Fusarium moniliforme and Fusarium oxysorum. This study supports the folkloric claims for the use of Gmelina arborea against some bacterial caused infections.

Polyphenol composition and antioxidant activity of Searsia tripartita and Limoniastrum guyonianum growing in Southeastern Algeria

Due to the notable medicinal value of Searsia tripartita and Limoniastrum guyonianum, it was of great interest to conduct phytochemical and antioxidant investigations. Polyphenol extracts of the dried powder of plants were prepared by maceration in 70% of methanol. The concentrated extracts were successively fractioned with distilled water, ethyl acetate and n-butanol. Obtained extracts were analysed for their phenol compositions by colorimetric processes and were evaluated for their total antiradical capacity by ABTS, DPPH and ORAC assays. Additionally, the phytochemical components were characterized in the methanolic extracts by HPLC–DAD–ESI-MS/MS in the negative ionisation mode. S. tripartita exhibited the higher quantity of phenolic compounds. The ethyl acetate extract of S. tripartita had the greatest quantities of polyphenols (55.5 ± 4.9 mg gallic acid equivalent per gram of dry residue (GAE/g DR)), flavonoids (44.2 ± 0.8 mg rutin equivalent per gram of dry residue (RE/g DR)), and tannins (12.5 ± 2.5 mg GAE/g DR). In L. guyonianum, the aqueous fraction had the highest quantity of polyphenols and tannins (10.2 ± 1.4 and 3.2 ± 1.9 mg GAE/g DR, respectively), whereas flavonoids (1.8 ± 0.2 mg RE/g DR) were higher in the ethyl acetate portion. The antioxidant capacity of the hydromethanolic extract of S. tripartita was found to be 3–16 times more effective than that of L. guyonianum using ORAC, DPPH, ABTS tests. HPLC analysis of hydromethanolic extracts provided tentative identification of four flavonoid glycosides in S. tripartita (myricetin-3-O-glucoside, myricetin-3-O-rhamnoside, quercetin-3-O-glucoside, and quercetin-3-O-rhamnoside) and five phenolic metabolites of the flavonoid class in L. guyonianum (myricetin-3-O-rhamnosylglucoside, myricetin-3-O-glucuronide, myricetin-3-O-pentoside, quercetin-3-O-glucuronide, and eriodictyol-7-O-rutinoside). Our findings revealed that these plants could be used as a potent source of health molecules.

Phytochemical Investigation and Antimutagenic Potential of Ethanolic Extracts of Emblica officinalis, Terminalia chebula and Terminalia bellirica

Background: Emblica officinalis Gaertn. which belongs to the family Euphorbiaceae, Terminalia chebula Retz. and Terminalia bellerica Roxb. belong to the family Combretaceae. These are well known medicinal plants with phytochemical reservoir of great medicinal values and possess a vast ethnomedical history. Objective: The aim of the present study is to isolation of major compounds and to evaluate antimutagenic potential of the ethanol extracts of these plants. Methods: The dried fruits of E. officinalis, T. bellirica and T. chebula were powdered and extracted with 95% ethanol. The ethyl acetate portions were chromatographed over silica gel to isolate major compounds. Antimutagenic activity was determined by Ames test using TA98 and TA100 strains of Salmonella typhimurium. Results: Two major known compounds, gallic acid and ellagic acid were isolated from the dried fruits of Emblica officinalis, Terminalia chebula and T. bellirica. All the three extracts counteracted the mutagenicity induced by different genotoxic compounds in a dose dependent manner. Conclusion: This study showed that ethyl acetate portion of three extracts contain two major compounds, gallic acid and ellagic acid which might be responsible for potent antimutagenic activity of these extracts.

Anti-epileptic potentials of the partitioned fractions of chamaecrista mimosoides

This research work aimed to establish scientific basis for the use of Chamaecrista mimosoides, in traditional medicine as anti-epileptic medication. The whole plant part of Chamaecrista mimosoides was extracted with ethanol and screened for phytochemicals. Acute toxicity study was carried out using Lorke’s method and the antiepileptic activity was evaluated using maximal electroshock induced seizure test in day-old chicks, pentylenetetrazole (PTZ) and strychnine using mice. The phytochemical study revealed the presence of saponins, cardiac glycosides, tannins, flavonoids, terpenoids and cardenolides. Both the chloroform, ethylacetate and n-butanol portions at 100, 250, and 500mg/kg body weight did not protect the chicks against tonic hind limb extension (THLE) in maximal electro-shock test (MEST). The chloroform and n-butanol portions at doses of 250 and 500 mg/kg body weight protected 40% and 60% of mice against clonic spasm induced by pentylenetetrazole, while ethyl-acetate soluble portion did not protect the mice against clonic spasm induced by pentylenetetrazole at all doses used when compared to Valproic acid (200 mg/kg) protected all the mice (100%) against clonic spasm induced by pentylenetetrazole. The chloroform soluble portion at the doses of 100, 250 and 500 mg/kg body weight protected 40%, 100%, 100% against death induced by strychnine, while ethylacetate and n-butanol portions did not protect the rats against death induced by strychnine but prolonged the onset of convulsion. In all the tests, phenobarbitone (20 mg/kg) was used as positive control and protected 80% of mice against convulsion induced by strychnine. The antiepileptic investigation suggests that the chloroform portion of Chamaecrista mimosoides has a promising antiepileptic activity. Â

Polyhydroxylated sterols from Monascus purpureus-fermented rice

Using a cell-based cytotoxicity assay, two new polyhydroxylated sterols, 16(S),22(S)-epoxy-3β,5α,6β,20(R),23(R),25-hexahydroxy-7-ergostene and 3β,7β,8α,25-tetrahydroxy-5,22E-ergostadiene were isolated from the ethyl acetate portion of the ethanolic extract of Monascus purpureus. Their structures were elucidated by spectroscopic analysis and in comparison with those reported in the literature. Both compounds showed cytotoxic activity against the lung adenocarcinoma (A549) with IC50 values of 12.6 and 18.5 μM, exhibited moderate activities against human ovarian cancer (A2780), with IC50 values of 8.8 and 9.4 μM.

A single-step isolation by centrifugal partition chromatography of the potential anti-inflammatory glaucolide B from Lepidaploa chamissonis

Sesquiterpene lactones (SL) are commonly found in Asteraceae and present a promising anti-inflammatory activity. Previously described in Lepidaploa genus, glaucolide B has never been investigated for its anti-inflammatory potential. This study aimed to establish an efficient process for the extraction of glaucolide B (1) from Lepidaploa chamissonis leaves and to develop a simple and fast method for its purification by using centrifugal partition chromatography (CPC), as well as to investigate in vitro the anti-inflammatory effects of glaucolide B. Thus, an optimized washing extractive process performed on L. chamissonis leaves allowed to obtain a SL enriched extract (4.11 g). After a successful defatting pretreatment of the crude extract, the glaucolide B enriched ethyl acetate portion (2.00 g) was fractionated by CPC affording, in a single-step isolation, compound 1 (1.04 g) in great yield (25%) and purity (97%). Cytotoxicity effect of 1 on RAW 264.7 macrophages was determined by using MTT assay, revealing a CC10 of 14.11 μM. Compound 1 at 1, 3 and 10 μM inhibited the nitrite/nitrate (NOx) metabolites production and the pro-inflammatory interleukin 6 (IL-6) secretion on lipopolysaccharide-stimulated RAW 264.7 cells. The extractive process used turned to be selective for SL and CPC technique proved a simple and effective tool for the isolation of 1 within few hours. Isolated for the first time from L. chamissonis leaves, glaucolide B presented a significant inhibitory effect on both NO and IL-6 secretion under non-toxic concentrations.

Isolation and Characterisation of Hydrolysable Tannin from Ethyl Acetate Portion of the Aerial Part of Phyllanthus amarus Schum. & Thonn

The present work involves extraction of phytochemicals from aerial part of Phyllanthus amarus Schum. & Thonn with n-hexane and 85% methanol. The isolation and characterization of Phytoconstituents was done from the methanol extract through portioning with chloroform and ethyl acetate. Fractionation and isolation (using column and thin layer chromatography respectively) of ethyl acetate column pooled portion afforded a compound coded as E-3.3C. The structure of the isolated compound was established on spectroscopic evidences (IR, 1HNMR, MS), which revealed the compound as 1-de (oxygalloyl)-2ˈ,3ˈ,-di-methoxy-amariin a hydrolysable tannins.

In vitro inhibition of a-amylase enzyme, phytochemical study and antioxidant capacity for cupressus sempervirens extracts growing in arid climat

Cupressus sempervirens (Cupressaceae) “cypress” is growing in the Mediterranean basin; it is a medicinal and an aromatic plant it used in treatment of many diseases in traditional medicine. This paper aims to determining antioxidant potential of C. sempervirens extracts and evaluating their inhibitory effects on α-amylase enzyme. The antioxidant properties of the crude extract and its factions were assessed by phosphomolybdenum, DPPH assay and α-amylase inhibitory activity was evaluating by spectrophotometric methods. Results showed that all extracts have high reductive activity better than butylated hydroxyanisole (BHA) with Ascorbic Acid Equivalent Anti-oxidant Capacity (AEAC) values ranged between 789.04 ± 32.47 and 28.18 ± 3.09 mM, while the greatest radical scavenging activity was found at ethyl acetate fraction with IC50 value 10 μg/ml, whereas in α-amylase enzyme inhibitory activity, ethyl acetate fraction of fruits showed the highest α-amylase inhibitory activity (43.35%). While the inhibitory patterns of ethyl acetate fraction of fruits (EFF) and ethyl acetate portion of tannin extract for leaves (EFT) were competitive type.

Anti-inflammatory active fraction screening and mechanism of unripe Forsythiae Fructus based on ¹H-NMR metabolomics

To screen the anti-inflammatory active fraction of unripe Forsythiae Fructus, and elucidate the action mechanism, water decoction, ethyl acetate portion, n-butanol portion and residue water extracts of unripe Forsythiae Fructus were administered into rats for continuously 15 days. The acute lung injury inflammatory model was established to observe the section structure of lung tissues. Levels of IL-6, TNF-α, IL-1β and IL-10 in bronchoalveolar lavage fluid were determined by ELISA kits, and changes in endogenous metabolites in serum were analyzed based on 1H-NMR metabolomics. The results showed that ethyl acetate portion of unripe Forsythiae Fructus had a better anti-inflammatory activity against acute lung injury, and could suppress the release of inflammatory factors of IL-6, TNF-α, IL-1β, significantly reduce contents of creatine, β-OH-butyrate, succinate, lysine, valine, isoleucine and glutamine, and elevate the content of GPC in serum. Ethyl acetate portion was proved to be the main fraction of anti-inflammatory activity from the perspective of endogenous metabolites in serum, and played an anti-inflammatory role by regulating creatine metabolism, choline metabolism, branched-chain amino acid metabolism and TCA cycles. This study could lay a foundation for studying pharmacodynamic material basis of unripe Forsythiae Fructus.

(+)-Spectaline, a Piperidine Alkaloid from <i>Senna spectabilis</i> DC. Effective in Reducing the <i>In Vitro</i> Infection of <i>Leishmania major</i>

Senna spectabilisis known to have antimicrobial, laxative, antiulcerogenic, analgesic, and anti-inflammatory properties in folk medicine. Piperidine alkaloids extracted from various parts of this plant have been shown to have anticonvulsant (iso-6-spectaline), antinociceptive [(-)-spectaline] and lipid peroxidation [(+)-3-O-feruloylcassine, (-)-spectaline and (-)-3-O-acetylspectaline] activities. In our study, the ethyl acetate extract fromS. spectabilisexhibited antileishmanial activity via intracellular promastigote assay or leishmanicidal assay and was further fractionated by using bioassay-guided isolation approach. The antiprotozoal principle was isolated from the ethyl acetate portion through solvent fractionation and a few series of chromatographic processes. The isolated active compound 1 was identified as (+)-spectaline on the basis of its spectral analysis (MS, 1D & 2D NMR) with EC50value of 0.063 ± 0.005 µM for antileishmanial activity and selectivity index of 3.76.

Isolation and identification of a new neo-clerodane diterpenoid from Teucrium chamaedrys L.

Teucrium chamaedrys L. is an aromatic and medicinal plant used as traditional medicine. Aerial parts of the plant material were dried and extracted with hexane–dichloromethane (extract 1), ethyl acetate–dichloromethane (extract 2) and methanol–dichloromethane (extract 3) in a ratio of 1:1 at rt successively. The solvents were evaporated to give crude extracts. Extract 1 was suspended in water at 60°C then partitioned successively with hexane and ethyl acetate to give hexane and ethyl acetate portions. After the column chromatography (silica gel) of ethyl acetate extract, one new and four known compounds were isolated. The new compound was named as 1(12S,18R)-15,16-epoxy-2β,6β-dihydroxy-neo-cleroda-13(16),14-dien-20,l2-olide-l8,l9-hemiacetal (teuchamaedryn D) (4). The known compounds were teucrin A (1), dihydroteugin (2), teucroxide (3), syspirensin A (5). The chromatographic methods were also applied for extract 3 to isolate verbascoside (6) and teucrioside (7). The structure of isolated compounds was elucidated by spectroscopic methods including LC-TOF/MS, 1D NMR and 2D NMR.