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Introduction: Toddalia asiatica (TA) is a classical traditional Chinese medicine used to treat rheumatoid arthritis and contusions. However, research regarding TA quality control is currently limited. Objective: We aimed to establish a strategy for identifying quality markers that can be used for the evaluation of the quality of TA. Method: A rapid and efficient ultra‐high‐performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC‐MS/MS) method was developed for the quantitative determination of 19 compounds in TA from different regions. Then, the extraction process of TA was successively optimized by single‐factor optimization and response surface methodology. Moreover, chemometrics was employed to confirm the correlation between quality and target compounds. Results: Utilizing the UHPLC‐MS/MS method, separation of the 19 bioactive compounds was achieved within 14 min. The method was validated in terms of linearity (r2 > 0.9982), precision (0.08%–3.70%), repeatability (0.50%–2.54%), stability (2.26%–5.46%), and recovery (95.8%–113%). The optimal extraction process (extraction solvent, 65% ethanol aqueous solution; solid–liquid ratio, 1:20; extraction time, 25 min) was determined with the total content of 19 bioactive compounds as indicator. Significant disparities were observed in the contents of target compounds across different batches of TA. Besides, all samples could be categorized into two distinct groups, and magnoflorine, (−)‐lyoniresinol, nitidine chloride, norbraylin, skimmianine, and decarine were identified as quality markers. Conclusion: In the present study, we developed a strategy to improve the quality control of TA. In consideration of the pharmacodynamic activity and statistical differences, six compounds are proposed as quality markers for TA. A rapid and efficient UHPLC‐MS/MS method was developed for the quantitative determination of 19 compounds in Toddalia asiatica from different regions. All samples could be categorized into two distinct groups, and magnoflorine, (−)‐lyoniresinol, nitidine chloride, norbraylin, skimmianine, and decarine were identified as quality markers. A new strategy of UHPLC‐MS/MS with chemometrics was developed to improve the quality control of Toddalia asiatica. [ABSTRACT FROM AUTHOR]

A new prenylated coumarin, 5, 7-dimethoxy-6-[(Z)-3'-methylbutan-1',3'-d-ienyl] coumarin (1), along with ten known compounds (2-11), was obtained from the roots of Toddalia asiatica (Linn.) Lam. Their structures were determined based on 1D and 2 D NMR spectroscopy as well as high-resolution mass spectrometry. Notably, the structure of (3S, 4R)-3,4-epoxypimpinellin was revised as its cis-epimer, compounds (2-6) were isolated from this genus for the first time. The cytotoxic activities of the isolated compounds 1-11 were evaluated against SW480, A549, MDA-MB-231, HepG 2, HEp-2, SGC7901 cancer cell lines. Compound 10 exhibited weak cytotoxicity against HEP 2 with IC50 value of 75.19 ± 1.17 µM. [ABSTRACT FROM AUTHOR]

Human hormone glucagon-like peptide-1 (GLP-1) lowers α-cell glucagon and increases β-cell insulin. GLP-1 is digested by dipeptidyl peptidase (DPP-4) enzyme; hence, the inhibition of DPP-4 enzyme prevents the digestion of GLP-1 and reduces the occurrence of type 2 diabetes condition. Present study developed a database consists of 1837 molecules structurally similar to selected query compounds applying PubChem screening and was further screened for drug-likeness and ADME/T behaviour using SwissADME tool. Total of six compounds, i.e. Toddalolactone, Ulopterol, 1-O-methyl 4-O-(2-oxo-1H-quinolin-4-yl) butanedioate, [2-oxo-3-(3-oxobutyl)-1H-quinolin-4-yl] acetate, N-(2,5-Dioxo-5,6-dihydro-2H-pyrano[3,2-c]quinolin-3-yl)acetamide, (2S)-2-amino-2-phenyl-1-[3-(trifluoromethyl)-6,8-dihydro-5H-[1,2,4] triazolo [4,3-a]1yrazine-7-yl] ethanone cleared screening and were tested for DPP-4 enzyme inhibitory activity comparing Alogliptin as standard drug applying AutoDock in silico approach (Argus Lab, Discovery Studio and PyMol). All the six compounds displayed RMSD values less than 2A0, and binding energies of − 8.5139 kcal/mol, − 9.438 kcal/mol, − 9.4783 kcal/mol, − 8.898 kcal/mol, − 9.022 kcal/mol, and − 7.604 kcal/mol, respectively. Study testified that Toddalolactone isolated from the plant Toddalia asiatica (− 8.5139 kcal/mol) confers 03 hydrogen bonds, 03 Pi-Pi stack bonds and RMSD values of 0.281 A0, phytocompound Ulopterol (− 9.438 kcal/mol) isolated from the plant Coleonema album, interacted with 04 amino acids of DPP-4 enzyme active site conferring stable hydrogen (02) and Pi-Pi bonds (02) with RMSD value 0.78 A0, represents increased DPP-4 enzyme inhibitory activity in comparison with standard drug Alogliptin binding energy of − 7.34 kcal/mol with only 02 interactive bonds. It is evidenced from the study that a higher number of bonds indicates the stability of the complex formed with increased enzyme inhibitory activity; hence, the phytocompound Toddalolactone derived from the plant Toddalia asiatica and Ulopterol from Coleonema album as a potential lead molecule shall be tested further resulting in in vitro and in vivo procedures in order to develop novel drugs in the treatment of Type 2 Diabetes mellitus condition. [ABSTRACT FROM AUTHOR]

AIM: Observation of neuroprotective effects of Toddalia asiatica (TA) on cerebral ischemia-reperfusion injury (CIRI) in rats by investigating the effects and mechanisms of drugs on the polarization of microglia M1/M2 subtype and the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor-κB (NF-κB) pathway. METHODS: The modified thread occlusion method was used to establish a rat model of CIRI, and the rats were randomly divided into the model group, Toddalia asiatica (1.08 g/kg) group, donepezil hydrochloride (0.45 mg/kg) group, and sham group, with 16 rats in each group. Based on the assessment of neurofunctional changes in each group of rats, HE and Nissl staining were used to observe the pathological changes in brain tissue. TUNEL staining was performed to detect neuronal apoptosis, immunohistochemistry was used to detect the expression of M1 microglia marker ionized calcium-binding adapter molecule 1 (Ibal), M2 microglia marker arginase 1 (Argl), and TLR4 localization. Western blot was used to detect the expression of microglia polarization proteins and proteins related to TLR4/MyD88/NF-κB pathway in the hippocampus region. RESULTS: Compared with sham group, the model group rats had higher neurological function scores (P<0.01), and neuronal arrangement in the hippocampus and cortex was loose and disordered, Nissl bodies decreased, and neuronal apoptosis increased. The numbers of M1 microglia marker Iba1-, M2 microglia marker Arg1-, and TLR4-positive cells were significantly increased. In addition, the protein levels of TLR4, MyD88, p-NF-κB p65, NF-κB p65, p-NF-κB inhibitory factor (p-IκB), Ibal, interleukin-6 (IL-6), and Arg1 in the hippocampus were elevated (P< 0.05, while IL-4 and IL-10 expression were decreased (P<0.01). Compared with model group, the Toddalia asiatica group and Donepezil hydrochloride group showed increased protein expression of Arg1, IL-4 and IL-10 (P<0.05), while the other indicators were decreased (P<0.05, P<0.01). CONCLUSION: Toddalia asiatica possesses neuroprotective effects on CIRI rats, which may be attributed to its ability to regulate M1/M2 polarization and inhibit the TLR4/MyD88/NF-κB-mediated inflammatory pathway. [ABSTRACT FROM AUTHOR]

Two new coumarins with a rare, oxidized geranyl group, named toddalins I (1) and J (2), were isolated from the roots of Toddalia asiatica , a traditional Chinese herb used to treat many diseases, especially athletic injuries. Their structures were determined using extensive spectroscopic methods, including 1D NMR, 2D NMR and HRESIMS. Toddalins I (1) and J (2) exhibited moderate in vitro anti-inflammatory activities by the inhibition of lipopolysaccharide (LPS)-induced NO production in RAW 264.7 macrophages with IC 50 values of 12.6 and 15.6 μ M, respectively. [Display omitted] • Two new coumarins (1 and 2) were isolated from the roots of Toddalia asiatica. • Their structures were elucidated by means of extensive spectroscopic analyses. • Toddalins I (1) and J (2) exhibited moderate in vitro anti-inflammatory activities. [ABSTRACT FROM AUTHOR]

We reported the discovery of six novel coumarins, toddasirins A–F (1 – 6), each endowed with modified isoprenyl or geranyl side chains, derived from the roots of Toddalia asiatica. Comprehensive structural elucidation was achieved through multispectroscopic analyses, single-crystal X-ray diffraction experiments, and advanced quantum mechanical electronic circular dichroism (ECD) calculations. Furthermore, the anti-inflammatory activity of these compounds was assessed. Notably, compounds 1 – 3 and 6 demonstrated notable inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 cells, with 50% inhibitory concentration (IC 50) values of 3.22, 4.78, 8.90, and 4.31 μmol·L−1, respectively. [ABSTRACT FROM AUTHOR]

The antifungal and antiaflatoxigenic effects of four distinct plant species against Aspergillus flavus and Aspergillus parasiticus were investigated. Essential oils and methanolic extracts were prepared from aerial parts of Lippia javanica, Ocimum gratissimum, Satureja punctata, and stem barks of Toddalia asiatica by hydro-distillation and maceration, respectively. The poisoned food method was used to confirm the antifungal activity of essential oils and methanolic extracts from four different plant species against Aspergillus flavus and Aspergillus parasiticus, and high-performance liquid chromatography was used to quantify the antiaflatoxigenic activity. The essential oils of Satureja punctata and Lippia javanica showed the highest antiaflatoxigenic activity against the fungi strains tested at concentrations of 1.25, 2.5, and 5 µL/mL, followed by Ocimum gratissimum essential oil while Toddalia asiatica essential oil exerted moderate antiaflatoxigenic activity. Meanwhile, the methanolic extracts showed a wide spectrum of low to high antifungal and antiaflatoxigenic activities at concentrations of 125, 250, and 500 µg/mL against A. flavus and A. parasiticus. This study has indicated that the essential oils of Satureja punctate, Lippia javanica, and Ocimum gratissimum had substantial antifungal and antiaflatoxigenic activities compared to their methanolic extracts, while Toddalia asiatica methanolic extract had a moderate antifungal activity compared to its essential oil. [ABSTRACT FROM AUTHOR]

Medicinal plants are rich sources of several diverse bioactive constituents with a wide range of biological effects. The screening of bioactive compounds plays a vital role in the protection of human health. Toddalia asiatica (L.) Lam., belonging to Toddalia genus of Rutaceae family, is a folk medicine in China used for hundreds of years. The purpose of the current study is to monitor the phytochemical compounds in the root of Toddalia asiatica using gas chromatography and mass spectrometry (GC–MS). Totally 27 compounds were identified and the chromatograph showed peaks with individual compounds. The major constituents identified in root methanolic extract were anliinosoquinoline (20.18%), pimpinellin (20.18%), pyrrolidine (18.92%), 1H-pyrrole (18.41%) and 2-phenyl-6-vinylindolizine (15.53%) and many other compounds were identified as low level. Many of these phytocomponents have antibacterial, antifungal, antifouling, immunomodulatory and antitumor activities, which justify the folklore use of root extract in traditional system to cure various ailments. [ABSTRACT FROM AUTHOR]

A new sesquiterpene, named todasinoid A (1), together with eight known compounds (2−9) were isolated from the roots of Toddalia asiatica. The gross structure of todasinoid A were established by analyses of the NMR and HRESIMS data. A comparison of the experimental ECD spectrum of 1 with the calculated ECD spectra for a model compound (1a) resolved the absolute configuration of 1. The known compounds were identified to be bullatantriol (2), aculeatin (3), 6-(3-ethoxy-2-hydroxy-3-methylbutyl)-5,7-dimethoxy-2H-1- benzopyran-2-one (4), trans-N-p-coumaroyltyramine (5), feruloyltyramine (6), −sitosterol (7), sitosterol Dglucoside(8), 7α-hydroxysitosterol (9) by comparing the NMR data and specific rotations with reported data in literature. Compound 1 is an eremophilane-type sesquiterpenoid containing a mercaptolactate side-chain that is rarely found in nature. Compounds 2, 5, and 9 were isolated from this plant for the first time. Bioassay study revealed that compound 5 exhibited inhibitory effects against -glucosidase with an IC50 of 320 M, being more active than the positive control acarbose. [ABSTRACT FROM AUTHOR]

Toddalia asiatica (L.) Lam., belonging to Toddalia genus of Rutaceae family, is a folk medicine in China used for hundreds of years. The whole plant can be used as medicine, especially the root that used to be applied in the folk. In recent decades, with the in-depth research from domestic and foreign researchers, it has gradually been discovered that the chemical components in T. asiatica are mainly coumarins and alkaloids. Its pharmacological effects are manifested in anti-inflammatory and analgesic, hemostatic coagulation, anti-tumor, treatment of cardiovascular diseases, etc. It has a wide range of clinical applications and significant effects on rheumatism, pain, wound bleeding, and bruises. Due to its important research value, in this article, the chemical compositions and pharmacological effects of T. asiatica are comprehensively expounded in recent years in order to provide a reference for the related research and application of this medicinal material, which were carried out through a bibliometric search using the Science Citation Index- Expanded (SCIE) database, web of science, Google scholar and Chinese National Knowledge Infrastructure (CNKI) and all that.

The present study reports the enhanced antifungal, aflatoxin B 1 (AFB 1) inhibitory activities and mode of action of chitosan-based nanoencapsulated Toddalia asiatica essential oil (neTAEO). Twenty-seven different chemical components were recognized from T. asiatica essential oil (TAEO) using gas chromatography–mass spectrometry (GC–MS). The caryophyllene oxide (CO) (25.4%), and 1,3-hexadiene, 3-ethyl-2,5-dimethyl- (HED) (24.08%) were documented as significant compounds. The Z -average particles diameter (Z-APD) of the neTAEO ranged between 18.41 and 131.8 nm. The neTAEO showed enhanced and most promising antifungal and AFB 1 inhibitory activity than TAEO. In viable maize model assay, neTAEO effectively preserved the maize from fungal invade and AFB 1 biosynthesis. The neTAEO significantly disturbs membrane integrities of Aspergillus flavus by inhibiting ergosterol biosynthesis followed by the extreme release of ions (Mg2+ and K+) and UV-absorbing (260 and 280 nm) cellular constituents. The in-silico molecular docking showed that the major active components of TAEO viz. , CO and HED were active against AFB 1 synthesizing leading genes Ver-1 and Omt-A with docking scores ranging from −4.8 to −7.7. The obtained results confirm that neTAEO showed promising antifungal and AFB 1 inhibitory activities; hence, it could be used as an alternative green strategy to protect food grains from fungal invade and AFB 1 production during storage. [ABSTRACT FROM AUTHOR]

Foods contaminated with fungi and their toxins cause many infections in developing and underdeveloped countries. Owing to the development of resistance by fungal pathogens to existing chemical antifungal agents, there is an urgent need to look for new eco-friendly antimicrobial agents. There is an increasing demand from agro-food industries for new environmentally compatible fungicides. Plant essential oils are an excellent source of antifungal volatile substances which are appropriate substitutes for food protection. Keeping this in mind we evaluated the efficacy of selected essential oils in suppressing the growth of Alternaria alternata isolated from stored wheat grains using in vitro and in vivo models. Preliminary antifungal activities varied among the tested oils with the zones of inhibition ranging from 10.66±1.15 mm to 19±1.00 mm. Among the tested oils T. asiatica oil showed significant inhibitory effects on A. alternata at 2.5 µL/mL. Further, its mechanistic action was related to the impairment of the squalene gene (SqSy) which was involved in the biosynthesis of sterol and ergosterol membrane factors. This was further supported by altered sterol profile of A. alternata with 29.4±2.40% reduction of total ergosterol content at 30 µL/mL. Moreover, prolonged exposure to T. asiatica oil as a fumigant controlled the total fungal count up to 68.75% compared to the control for a period of 12 months. T. asiatica oil proved to be a promising candidate to control A. alternata and its mycotoxin residues in stored wheat. It can be used as a sustainable and effective alternative to develop new fungicide. [ABSTRACT FROM AUTHOR]

Medicinal plants have become important sources of natural products, which have been used in the development of therapeutic agents. Four new coumarins (1-4) have been isolated together with five known metabolites (5-9) from the medicinal plant T. asiatica. The structures of 1-9 were assigned based on their spectroscopic data. Compounds (1-9) inhibited the growth of the Gram-negative bacteria E. coli and Gram-positive bacteria S. aureus at a concentration of 25 and 50 μg/disc. Compounds (2-4) inhibited the phytopathogenic fungus C. cucurbitarum at 50 μg/disc.

Objective: To investigate the therapeutic mechanism of Tujia medicine Toddalia asiatica alcohol extract (TAAE) for synovial pannus formation in rats with college-induced arthritis (CIA)., Methods: Sixty male SD rats were randomized into normal control group, CIA model group, TGT group, 3 TAAE treatment groups at low, medium and high doses ( n =10). Except for those in the normal control group, all the rats were subjected to CIA modeling using a secondary immunization method and treatment with saline, TGT or TAAE by gavage once daily for 35 days. The severity of arthritis was assessed using arthritis index (AI) score, and knee joint synovium pathologies were examined with HE staining. Serum levels of TNF-α, IL-6, and IL-1β were detected with ELISA; the protein expressions of PI3K, Akt, p-PI3K, p-Akt, VEGF, endostatin, HIF-1α, MMP1, MMP3, and MMP9 in knee joint synovial tissues were determined using Western blotting, and the mRNA expressions of TNF‑α, IL-6, IL-1β, VEGF, HIF-1α, PI3K, and Akt were detected with RT-PCR., Results: Treatment of CIA rat models with TAAE and TGT significantly alleviated paw swelling, lowered AI scores, and reduced knee joint pathology, neoangiogenesis, and serum levels of inflammatory factors. TAAE treatment obviously increased endostatin protein expression, downregulated p-PI3K, p-Akt, MMP1, MMP3, MMP9, VEGF, and HIF-1α proteins, and reduced TNF‑α, IL-6, IL-1β, PI3K, Akt, VEGF, and HIF-1α mRNA levels in the synovial tissues, and these changes were comparable between high-dose TAAE group and TGT group., Conclusion: TAAE can improve joint symptoms and inhibit synovial pannus formation in CIA rats by regulating the expressions of HIF-1α, VEGF, endostatin, MMP1, MMP3, and MMP9 via the PI3K/Akt signalling pathway.

Toddalia asiatica (L.) Lam., belonging to Toddalia genus of Rutaceae family, is a folk medicine in China used for hundreds of years. The whole plant can be used as medicine, especially the root that used to be applied in the folk. In recent decades, with the in-depth research from domestic and foreign researchers, it has gradually been discovered that the chemical components in T. asiatica are mainly coumarins and alkaloids. Its pharmacological effects are manifested in anti-inflammatory and analgesic, hemostatic coagulation, anti-tumor, treatment of cardiovascular diseases, etc. It has a wide range of clinical applications and significant effects on rheumatism, pain, wound bleeding, and bruises. Due to its important research value, in this article, the chemical compositions and pharmacological effects of T. asiatica are comprehensively expounded in recent years in order to provide a reference for the related research and application of this medicinal material, which were carried out through a bibliometric search using the Science Citation Index- Expanded (SCIE) database, web of science, Google scholar and Chinese National Knowledge Infrastructure (CNKI) and all that. [ABSTRACT FROM AUTHOR]

Ethyl acetate fraction of Toddalia asiatica was fractionated to yield fifteen previously undescribed prenylated coumarins, asiaticasics A-O (1 – 15) along with nine (16 – 24) known derivatives. The structures of these undescribed coumarins were established by spectroscopic analysis and reference data. Biological activity evaluation showed that compound 3 with the IC 50 value of 2.830 μM and compound 12 with the IC 50 value of 0.682 μM owned anti-inflammatory activity by detecting the rate of lactate dehydrogenase release in pyroptosis J774A.1 cells. The results showed that the expression of Caspase-1 and IL-1 β was decreased in a dose-dependent manner in the compound 12 treatment group, suggesting that compound 12 may reduce pyroptosis by inhibiting NLRP3 inflammasome. To further determine that compound 12 treatment can inhibit macrophage pyroptosis, morphological observation was performed and the results were consistent with the bioactivity evaluation. [Display omitted] • Twenty-four coumarins were isolated from the ethyl acetate fraction of Toddalia asiatica. • Fifteen coumarins that have not been described before. • Two undescribed compounds showed low IC 50 values and could exert effective anti-inflammatory effects. • Compound 12 inhibited pyroptosis in a dose-dependent manner. [ABSTRACT FROM AUTHOR]

This study aims to investigate the effect of Toddalia asiatica extract combined with miR-483 on osteoarthritis chondrocyte proliferation and apoptosis and expression of inflammatory factors. Osteoarthritis chondrocytes were treated with Toddalia asiatica extract. Osteoarthritis chondrocytes were transfected with anti-miR-483 or miR-483, and Toddalia asiatica extract was applied. The levels of TNF-α, IL-6, IL-10 were determined by ELISA. Cell proliferation and cloning were evaluated by MTT and cloning experiment. Cell apoptosis was determined by flow cytometry. P21 and caspase-3 protein expression were analyzed by Western blot. The expression of miR-483 was detected by qRT-PCR. The application of Toddalia asiatica extract or inhibition of miR-483 significantly increased the cell survival rate, cloning count and IL-10 level of osteoarthritis chondrocytes, and significantly reduced the apoptosis rate, levels of P21, Caspase-3, TNF-α IL-6 and miR-483 expression level of osteoarthritis chondrocytes (p<0.05). Overexpression of miR-483 could reverse the promotion effect of Toddalia asiatica extract on osteoarthritis chondrocytes proliferation, clone formation and IL-10 level, as well as reserve the inhibition effect of Toddalia asiatica extract on osteoarthritis chondrocyte apoptosis and levels of P21, caspase-3, TNF-α, IL-6. The combination of Toddalia asiatica extract and miR-483 can promote the proliferation of osteoarthritis chondrocytes and inhibit apoptosis and expression of inflammatory factors. [ABSTRACT FROM AUTHOR]

A new sesquiterpene (1) and nine known alkaloids (2-10) were isolated from the roots of Toddalia asiatica. The structure of compound 1 were resolved by NMR and HRESI data, as well as ECD calculation for determining the absolute configuration. The known compounds were identified to be 8-acetonyldihydronitidine (2), 8-acetonyldihydroavicine (3), dihydronitidine (4), oxynitidine (5), decarine (6), skimmianine (7), γ-fagarine (8), N-methylflindersine (9), and 4-methoxy-N-methyl-2-quinolone (10) by comparing the NMR data with those in the literature. Compound 1 is the first eremophilane-type sesquiterpenoid isolated from this species. The known compounds 2, 3, and 6 were discovered for the first time from the genus Toddalia. All the isolated compounds were evaluated for their inhibitory effects against phosphodiesterase-4, as result, compound 2 showed strong inhibitory effect against phosphodiesterase-4 with an IC50 value of 5.14 μM. [ABSTRACT FROM AUTHOR]

Toddalia asiatica (L.) Lam. belongs to family Rutaceae and mainly distributes in dry areas of bushes in tropical Africa, Asia, and Swaziland. Sometimes it can be used as fodder for goats, but it has been used as herbs in traditional medical treatment for 1000 years. In this study, we sequenced the sample of T. asiatica and determined its complete chloroplast genome. The length of CP genome is 158,434 bp, includes two invert repeats (IR) regions of 27,008 bp, a large single-copy (LSC) region of 86,132 bp, and a short single-copy (SSC) region of 18,286 bp. There are 133 genes, which includes 88 protein-coding genes, 8 rRNA and 37 tRNA, and 38.5% overall GC content. Each of trnK-UUU, rps16, trnG-UCC, atpF, rpoC1, trnL-UAA, trnV-UAC, petB, petD, rpl16, rpl2, ndhB, trnI-GAU, trnA-UGC, and ndhA genes contains a intron, clpP and ycf3 contains 2 intron. The phylogenetic analysis result shows that T. asiatica has the closest relationship with Zanthoxylum armatum (MT990984) and Zanthoxylum nitidum (MN508801).

• ODCE, a CMC expansion strategy, was developed. • CMC was extended unidirectionally. • ODCE was compatible with the concurrent determination of multiple compounds. • Improvement of ULOQ reached 50-fold. To facilitate the safety, efficacy and rationality of clinical application of traditional Chinese medicines (TCMs), pharmacokinetic research played an indispensable role. The key challenge during pharmacokinetic investigation lied at the substantial fluctuation of compound concentrations in the plasma over the course of absorption. Taking the pharmacokinetics of six compounds after administration of Toddalia asiatica (TA) as an example, an efficient strategy was established by introducing the online double collision energy (ODCE) into the quantification process applying ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). During the analytical program, double collision energy (DCE) was optimized to establish the dual calibration curve (DCC) with large concentration monitoring coverage (CMC) for meeting the wide content range of certain target compounds. Method validation test was performed in terms of linearity, precision, sensitivity, matrix effect, recovery, etc. The results displayed that the CMC of todarolactone with high exposure in plasma was extended from 1.25–2,500 ng/mL to 1.25–125,000 ng/mL. Furthermore, a rapid UHPLC-MS/MS method integrated with ODCE was successfully applied to the determination of six compounds in rat plasma, revealing an extremely high plasma concentration of todarolactone (16,662 ng/mL). This strategy could expand the range of quantification while retaining extraordinary sensitivity. Consequently, it could be a fit-for-purpose strategy to quantify compounds over a wide concentration range for in vivo process monitoring. [ABSTRACT FROM AUTHOR]

Introduction: Coumarin and alkaloids are the major bioactive constituents of Toddalia asiatica, playing an important role in various biological activities such as anti‐inflammatory, analgesic, anti‐bacterial and anti‐tumour. Objective: To establish a method that will simultaneously determine the coumarins and alkaloids compounds in T. asiatica and identify their characteristic fragmentation patterns, while combining fingerprints and chemical identification with chemometrics for discrimination and quality assessment of T. asiatica samples. Methodology: Qualitative characterisation of coumarins and alkaloids compounds in the methanol extracts of T. asiatica was determined by ultra‐high‐performance liquid chromatography‐quadrupole time‐of‐flight tandem mass spectrometry (UPLC‐QTOF‐MS/MS). Quantitative analysis relies on high‐performance liquid chromatography with a diode array detector (HPLC‐DAD). Results: A total of 59 components were characterised by UPLC‐QTOF‐MS/MS, including 29 coumarin, 25 alkaloids, one phenolic acid and four flavonoids. While the 19 characteristic components out of 23 common peaks in the chromatographic fingerprints of T. asiatica were confirmed. Quantitative analysis of seven major compounds from 18 samples were simultaneously detected by HPLC‐DAD at wavelengths of 280 nm. The samples were classified into three groups by hierarchical clustering analysis (HCA) combined with principal component analysis (PCA), and orthogonal partial least squares discriminant analysis (OPLS‐DA) which screened out the main chemical markers responsible for the samples differences. Conclusion: Fingerprints combined with chemometrics and chemical identification are a simple, rapid and effective method for the quality control of T. asiatica. HighlightsA total of 59 compounds in T. asiatica were firstly identified.Seven major compounds in T. asiatica samples were simultaneously detected for the first time, which is meaningful for sample discrimination.By comparing with the reference substances and UPLC‐QTOF‐MS/MS data, 19 characteristic components out of 23 common peaks in the chromatographic fingerprints of T. asiatica were confirmed. [ABSTRACT FROM AUTHOR]

Coumarins and flavonoids are the major constituents of Toddalia asiatica. The separation and purification of ingredients from T. asiatica is an important procedure to acquire high-purity compounds for subsequent pharmacological investigation to discover leading compounds. In the present work, an offline two-dimensional high-performance liquid chromatography (HPLC) method was successfully established for the separation of high-purity glycosides from T. asiatica. Based on the separation results obtained with two different chromatographic stationary phases, a phenyl-bonded silica-based reversed-phase column was employed as the first HPLC preparation, and three fractions were obtained from the sample. Then, the fractions were isolated and purified on an octadecyl-bonded silica-based reversed-phase column to obtain high-purity compounds in the second HPLC separation. As a result, three coumarin glycosides, including two undescribed and one known, along with one known flavonoid glycoside with more than 98% purity were isolated from the sample. The structures of the isolated compounds were elucidated on the basis of extensive spectroscopic evidence derived from optical rotation, mass spectrometry, and nuclear magnetic resonance experiments. Two-dimensional HPLC with different stationary phases has the potential to be an efficient method for the separation of high-purity compounds from T. asiatica.

The EtOAc and MeOH extracts of the roots of Toddalia asiatica Lam. were investigated for the roots' chemical constituents. Two new compounds including 2′R-acetoxytoddanol (1) and 8S-10-O-demethylbocconoline (3) as well as 15 known compounds were isolated. Compound 10 showed strong cytotoxicity against KB cells with an IC50 value of 2.60 μg/mL, which is nearly equal to the ellipticine standard, but showed no activity against Vero cells. Alkaloid 3 displayed weak cytotoxicity against the KB cell line with an IC50 value of 21.69 μg/mL. [ABSTRACT FROM AUTHOR]

This study was aimed at evaluating the antibacterial prospective of essential oil (EO) extracted fromToddalia asiaticaL. against four pathogenic bacteria. The essential oil was steam-distilled using a modified Clevenger-type apparatus and its chemical composition established by gas chromatography mass spectrometry. Disc diffusion method was used to assess the antimicrobial activity of the essential oil against the test pathogens while tube dilution method was used for the determination of minimum inhibitory concentration and minimum bactericidal concentration of the EO on the test bacteria. The highest activity of the EO was observed againstE. coliwhere a mean inhibition zone of 34.00 mm was recorded at a concentration of 100% followed byB. cereuswhere a mean inhibition zone of 32.33 mm was recorded at an oil concentration of 50%. The minimum inhibitory concentrations and minimum bactericidal concentrations of the EO on the test pathogens were in the range of 3.13 12.25% and 12.25 >50%, respectively. When compared to tetracycline which was used as the standard, crude essential oil ofT. asiaticahad significantly higher activity in three out of four studied bacterial strains. These findings justify the continued use ofT. asiaticaextracts in traditional herbal medicine for the treatment of bacterial-based infections. [ABSTRACT FROM AUTHOR]

Bacteria have developed multidrug resistance against available antimicrobial agents. Infectious diseases caused by these multidrug-resistant bacteria are major causes of morbidity and mortality in human beings. Synthetic drugs are expensive and inadequate for the treatment of diseases, causing side effects and ineffective against multidrug-resistant bacteria. The medicinal plants are promising to have effective antimicrobial property due to presence of phytochemical compounds like alkaloids, flavanoids, tannins and phenolic compounds. The present study aimed to find the antimicrobial activity of medicinal plants against multidrug-resistant bacteria. Multidrug-resistant bacteria were identified by Kirby-Bauer disc diffusion method. Production of β-lactamases (extended spectrum β-lactamases, metallo β-lactamase and AmpC β-lactamase) were identified by combination disc method. Antibacterial activity of aqueous and ethanol extract of Aristolochia indica and Toddalia asiatica were detected by agar well diffusion assay and minimum inhibitory concentration. All bacteria used in this study showed antibiotic resistance to ≥3 antibiotics. Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus mirabilis and Vibrio cholerae were found to be positive for β-lactamase production. Ethanol extract of Aristolochia indica showed more significant antibacterial activity against multidrug-resistant bacteria than Toddalia asiatica. Ethanol extracts of Aristolochia indica and Toddalia asiatica showed minimum inhibitory concentration values of 50-100 μg/ml and 100-200 μg/ml, respectively against multidrug-resistant bacteria. From this study, it was concluded that Aristolochia indica has more potential to treat multidrug-resistant bacteria than Toddalia asiatica. [ABSTRACT FROM AUTHOR]

A chemical investigation of the ethanol extract from the roots of Toddalia asiatica resulted in the isolation of three new amides, (7Z )-N -(4′-methoxyphenethyl)-3-methoxy-4-hydroxycinnamamide ( 1 ), (7Z )-N -(4′-hydroxyphenethyl)-3-methoxy-4,5-dihydroxycinnamamide ( 2 ), and (7E )-N -(4′-hydroxyphenethyl)-3,4,5-trihydroxycinnamamide ( 3 ), together with one known amide (7Z )-N -(4′-hydroxyphenethyl-3-methoxy-4-hydroxycinnamamide ( 4 ). Their structures were elucidated on the basis of extensive 1D and 2D NMR (COSY, HMQC, HMBC, and NOESY) analyses. [ABSTRACT FROM AUTHOR]

INTRODUCTION: Toddalia asiatica is a commonly used medicinal plant in East Africa for the management of pain and inflammatory conditions. The present study investigated the antinociceptive and the anti-inflammatory effects of T. asiatica in Swiss albino mice. METHODS: The antinociceptive and the anti-inflammatory effects of T. asiatica were investigated using formalin-induced pain test and the carrageenin-induced oedema paw. The extract solvent (vehicle), aspirin and indomethacin were employed as negative and positive controls respectively. Eight mice were used in each experiment. RESULTS: In the early phase of the formalin test, the 100mg/kg dose showed no significant antinociceptive activity while the 200mg/kg showed significant (p 0.01) antinociceptive activity. The 100 mg/kg dose showed highly significant antinociceptive activity (p 0.001) in the late phase of the formalin test while the 200mg/kg dose showed no significant antinociceptive activity. A reduction in carragenin induced acute inflammation paw oedema was significant (p 0.01) following administration of 100mg/kg dose but not with the 200mg/kg dose.CONCLUSION: The present study therefore lends support to the anecdotal evidence for use of T. asiatica in the management of painful and inflammatory conditions.

Background: Toddalia asiatica (Rutaceae) is an important medicinal plant in traditional medicinal system of India and China. Nitidine production from callus cultures of the plant had been investigated, but in vitro multiplication and secondary metabolite production from shoot cultures is not reported. Objective: The aim of the present work is to establish protocol for in vitro multiple shoot regeneration of T. asiatica and to investigate the secondary metabolite, nitidine production from the shoot cultures. Materials and Methods: Different explants were used for shoot regeneration on MS supplemented with benzyl adenine (BA) either alone or in combination with naphthalene acetic acid (NAA) in different combinations. Effect of different sugars and different concentrations of sucrose on biomass accumulation in shoot cultures in liquid medium was investigated. For in vitro rooting, shoots culture were inoculated to half strength MS medium supplemented with different concentrations of indole butyric acid. Quantitative analysis of shoot culture extracts was done for estimation of nitidine by HPTLC. Results: Shoot cultures were successfully initiated and established from nodal and shoot tip explants on MS medium supplemented with benzyl adenine and sucrose (3% w/v). Sucrose at a concentration of 3 % w/v was found to be optimum for growth and biomass accumulation. In vitro rooting of shoots was achieved on half strength MS medium supplemented with indole butyric acid 3 mg/l. Investigation of secondary metabolite production ability of the in vitro regenerated shoot cultures revealed their ability to biosynthesize nitidine. Conclusion: Shoot cultures were established and nitidine production has been observed. [ABSTRACT FROM AUTHOR]

Abstract: Phytochemical investigation of the ethanol extract from the roots of Toddalia asiatica resulted in the isolation of eight new alkaloids, 8-methoxynorchelerythrine (1), 11-demethylrhoifoline B (2), 8-methoxynitidine (3), 8-acetylnorchelerythrine (4), 8,9,10,12-tetramethoxynorchelerythrine (5), isointegriamide (6), 1-demethyl dicentrinone (7), and 11-hydroxy-10-methoxy-(2,3)-methylenedioxytetrahydroprotoberberine (8), together with 10 known alkaloids. Their structures were determined on the basis of spectroscopic analyses, including 1D-NMR, 2D-NMR, and HR-ESI-MS. The isolated components were evaluated in vitro for cytotoxic activities against eight tumor cell lines, antimicrobial activities against two Gram-positive bacteria and five Gram-negative bacteria, and antifungal activities against five pathogens. Benzo[c]phenanthridine and secobenzo[c]phenantridine alkaloids exhibited significant cytotoxic, antimicrobial and antifungal properties. [Copyright &y& Elsevier]

Abstract: The parasitic ciliate Ichthyophthirius multifiliis infests all species of freshwater fish and can cause severe economic losses in fish breeding. The present study aims to evaluate the antiparasitic activity of the active components from Toddalia asiatica against I. multifiliis. Bioassay-guided fractionation and isolation of compounds with antiparasitic activity were performed on the methanol extract of T. asiatica yielding two bioactive compounds: chelerythrine and chloroxylonine identified by comparing spectral data (NMR and ESI-MS) with literature values. Results from in vitro antiparasitic assays revealed that chelerythrine and chloroxylonine could be 100% effective against I. multifiliis at the concentration of 1.2mgL−1 and 3.5mgL−1, with the median effective concentration (EC50) values of 0.55mgL−1 and 1.90mgL−1 respectively. In vivo experiments demonstrated that fish treated with chelerythrine and chloroxylonine at the concentrations of 1.8 and 8.0mgL−1 carried significantly fewer parasites than the control (P <0.05). The acute toxicity (LC50) of chelerythrine for goldfish was 3.3mgL−1. [Copyright &y& Elsevier]

Toddalia asiatica (L.) Lam. belongs to family Rutaceae and mainly distributes in dry areas of bushes in tropical Africa, Asia, and Swaziland. Sometimes it can be used as fodder for goats, but it has been used as herbs in traditional medical treatment for 1000 years. In this study, we sequenced the sample of T. asiatica and determined its complete chloroplast genome. The length of CP genome is 158,434 bp, includes two invert repeats (IR) regions of 27,008 bp, a large single-copy (LSC) region of 86,132 bp, and a short single-copy (SSC) region of 18,286 bp. There are 133 genes, which includes 88 protein-coding genes, 8 rRNA and 37 tRNA, and 38.5% overall GC content. Each of trnK-UUU, rps16, trnG-UCC, atpF, rpoC1, trnL-UAA, trnV-UAC, petB, petD, rpl16, rpl2, ndhB, trnI-GAU, trnA-UGC, and ndhA genes contains a intron, clpP and ycf3 contains 2 intron. The phylogenetic analysis result shows that T. asiatica has the closest relationship with Zanthoxylum armatum (MT990984) and Zanthoxylum nitidum (MN508801). [ABSTRACT FROM AUTHOR]

Highlights: [•] Toddalia asiatica is used for treating various diseases. [•] In vitro antioxidant activity of T. asiatica leaves. [•] Hypolipidemic activity of T. asiatica on triton and high fat diet induced models. [•] T. asiatica showed good hypolipidemic activity. [ABSTRACT FROM AUTHOR]

From the n-hexane, ethyl acetate and methanol extracts of Toddalia asiatica root bark were isolated eight compounds ( 1- 8) which were identified on the basis of both spectroscopic and physical data as well as comparison with already published results. The crude extracts and isolated compounds showed moderate in vitro antiplasmodial activity against D6 (chloroquine-sensitive) and W2 (chloroquine-resistant) strains of Plasmodium falciparum. The extracts and isolates also exhibited larvicidal activities against Aedes aegypti and coumarins were identified as the active compounds. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: Toddalia asiatica (L) Lam. (Rutaceae) is a medicinal plant traditionally used in Kenya by many communities for the treatment of malaria and other ailments. All parts of the plant are claimed to have medicinal value, but the root bark in particular is believed to be more potent. Decoctions or infusions of the roots are taken orally to treat malaria, fever and stomach ache. Aim of the study: To evaluate antimalarial activity of aqueous and organic extracts prepared from Toddalia asiatica and determine in vitro and in vivo safety of the extracts. Materials and methods: Aqueous, ethyl acetate, hexane and methanol extracts were obtained from Toddalia asiatica root bark, fruits and leaves. In vitro antiplasmodial activity was done using chloroquine-sensitive (D6) and chloroquine-resistant (W2) Plasmodium falciparum strains and the concentration causing 50% inhibition of radioisotope incorporation (IC50) was determined. In vivo assay was done by administering mice infected with Plasmodium berghei four consecutive daily doses of the extracts through oral route following Peters 4-Day suppressive test. The percentage suppression of parasitaemia was calculated for each dose level by comparing the parasitaemia in untreated control with those of treated mice. Quinine hydrochloride was used as positive control while double distilled water or 20% Tween-80 was used as a negative control. In vivo acute toxicity was determined in mice using standard procedures. In vitro cytotoxicity assay was carried out using actively dividing sub-confluent Vero cells. Results: Inhibitory concentrations of ethyl acetate extract of Toddalia asiatica fruits showed high activity against chloroquine resistant (W2) strains of Plasmodium falciparum (IC50=1.87μg/ml), followed by root bark aqueous extract (IC50=2.43μg/ml). Tested in vivo against Plasmodium berghei, the fruit ethyl acetate extract (500mg/kg) and root bark aqueous extract (250mg/kg) reduced malaria parasitaemia by 81.34% and 56.8% respectively. Higher doses were found to be less effective in vivo. Acute toxicity and cytotoxictiy of the tested extracts, with the exception of hexane extract from the roots, showed LD50>1000mg/kg and CC50>100μg/ml respectively. Conclusions: The results obtained contribute to the validation of traditional use of Toddalia asiatica and provides in vivo and safety data of the plant extracts tested for the first time. Ethyl acetate extract of the fruits was active against chloroquine resistant Plasmodium falciparum as well as against Plasmodium berghei. These findings confirm the suitability of Toddalia asiatica as a good candidate for further tests to obtain a prototype for antimalarial medicine. [Copyright &y& Elsevier]

Introduction: Toddalia asiatica is a commonly used medicinal plant in East Africa for the management of pain and inflammatory conditions. The present study investigated the antinociceptive and the anti-inflammatory effects of T. asiatica in Swiss albino mice. Methods: The antinociceptive and the anti-inflammatory effects of T asiatica were investigated using formalin-induced pain test and the carrageenin-induced oedema paw. The extract solvent (vehicle), aspirin and indomethacin were employed as negative and positive controls respectively. Eight mice were used in each experiment. Results: In the early phase of the formalin test, the 100mg/kg dose showed no significant antinociceptive activity while the 200mg/kg showed significant (p < 0.01) antinociceptive activity. The 100 mg/kg dose showed highly significant antinociceptive activity (p < 0.001) in the late phase of the formalin test while the 200mg/kg dose showed no significant antinociceptive activity. A reduction in carragenin induced acute inflammation paw oedema was significant (p < 0.01) following administration of 100mg/kg dose but not with the 200mg/kg dose. Conclusion: The present study therefore lends support to the anecdotal evidence for use of T asiatica in the management of painful and inflammatory conditions. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: The leaves of Toddalia asiatica (L.) Lam. have been utilized traditionally for the cure of diabetes. Aim of the study: The present study was aimed to assess the antidiabetic and antioxidant effects of T. asiatica leaves in Streptozotocin (STZ) induced diabetic rats. Materials and methods: The phytochemical screening, total phenolic content, HPLC analysis, acute toxicity study and oral glucose tolerance test were carried out. Glucose lowering effect of the hexane, ethyl acetate and methanol extracts of T. asiatica leaves was studied in STZ-induced diabetic rats. The antidiabetic and antioxidant activities were studied for the ethyl acetate extract. The effects of extracts on blood glucose, body weight, plasma insulin, total protein, liver glycogen, plasma enzymes (SGOT, SGPT and ALP) and activities of SOD, CAT and GPx were analyzed. Results: T. asiatica leaves ethyl acetate extract (TALEe) showed highly significant blood glucose lowering effect. Phytochemical evaluation of TALEe showed the presence of alkaloids, terpenoids, cumarins, flavonoids and phenolic compounds. The total phenolic content of TALEe was 126mg of gallic acid equivalents/g extract. HPLC analysis revealed the presence of flindersine and ulopterol. Acute toxicity study of TALEe revealed no death or toxicity. The oral glucose tolerance test showed lowered area under curve (AUCglucose) values in TALEe treated rats. After treatment with TALEe (250 and 500mg/kg) for 28 days there was a significant decrease in blood glucose, plasma enzymes (SGOT, SGPT and ALP) and significant increase in body weight, total protein, serum insulin and liver glycogen levels in treated diabetic rats. The activities of antioxidant enzymes SOD, CAT and GPx were reversed to near normal in treated diabetic rats. Histopathology of pancreas in TALEe treated groups showed regeneration of β-cells. Conclusion: The results of the experiments showed that TALEe exerted significant antidiabetic and antioxidant effects in STZ-induced diabetic rats justifying its traditional use. [Copyright &y& Elsevier]

Abstract: Chemotherapeutics represent the main approach for the treatment of leukemia. However, the occurrence of adverse side effects and the complete lack of effectiveness in some cases make it necessary to develop new drugs. As part of our screening program to evaluate the potential chemotherapeutic effect of natural coumarins, we investigated the anti-leukemic activities of a series of six prenylated coumarins isolated from the stem bark of Toddalia asiatica (Rutaceae). Among these, 6-(3-methyl-2-butenyl)-5,7-dimethoxycoumarin (toddaculin) displayed the most potent cytotoxic and anti-proliferative effects in U-937 cells. To determine whether these effects resulted from induction of cell death or differentiation, we further evaluated the expression of several apoptosis and maturation markers. Interestingly, while toddaculin at 250μM was able to induce apoptosis in U-937 cells, involving decreased phosphorylation levels of ERK and Akt, 50μM toddaculin exerted differentiating effects, inducing both the capacity of U-937 cells to reduce NBT and the expression of differentiation markers CD88 and CD11b, but no change in p-Akt or p-ERK levels. Taken together, these findings indicate that toddaculin displays a dual effect as a cell differentiating agent and apoptosis inducer in U-937 cells, suggesting it may serve as a pharmacological prototype for the development of novel anti-leukemic agents. [Copyright &y& Elsevier]

Abstract: A rapid and selective ultra performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) method was developed for simultaneous determination of three bioactive coumarins of Toddalia asiatica extract including pimpinellin, isopimpinellin and phellopterin in rat plasma for the first time. Phenacetin was used as the internal standard (IS). Plasma samples were extracted by liquid–liquid extraction with methyl tert-butyl ether. The chromatographic separation was carried out on an ACQUITY UPLC™ BEH C18 column with an isocratic mobile phase consisting of methanol-5mmol/L ammonium acetate (65:35, v/v). The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source with positive ionization mode. The method was linear for all analytes over investigated range with all correlation coefficients greater than 0.9942. The lower limits of quantification (LLOQ) were 25.0ng/mL for pimpinellin, 10.0ng/mL for isopimpinellin and 5.00ng/mL for phellopterin. The intra- and inter-day precision (RSD%) was within 12% and the accuracy (RE%) ranged from −2.3% to 5.5%. The rapid and sensitive method was fully validated and successfully applied to the pharmacokinetic study of pimpinellin, isopimpinellin and phellopterin in rats following oral administration of Toddalia asiatica extract. [Copyright &y& Elsevier]

Abstract: Ethnopharmacological relevance: The leaves of Toddalia asiatica (L.) Lam. (Rutaceae) are widely used in folk medicine in India to treat various ailments like cough, malaria, indigestion, influenza lung diseases and rheumatism, fever, stomach ailments, cholera and diarrhea. In our earlier communication we have reported the antimicrobial study on the various extracts of the leaves and the isolation and identification of Flindersine, a quinolone alkaloid as the major active principle. In the present study, we report the antibacterial and antifungal activities of Ulopterol, a coumarin isolated as another major active antimicrobial principle. Materials and methods: The leaves were successively extracted with hexane, chloroform, ethyl acetate, methanol and water. The extracts were studied for their antimicrobial activity against selected bacteria and fungi by using disc-diffusion method. The ethyl acetate extract which was found to possess highest antimicrobial activity was subjected to activity guided fractionation by column chromatography over silica gel. This resulted in the isolation of the coumarin, Ulopetrol, an active principle besides Flindersine which was reported by us earlier. The structure of the compound was elucidated using physical and spectroscopic data. Flindersine and Ulopterol were quantified by HPLC. Results: Ulopterol showed activity against the bacteria viz. Staphylococcus epidermidis, Enterobacter aerogenes, Shigella flexneri, Klebsiella pneumoniae (ESBL-3967), Escherichia coli (ESBL-3984) and fungi viz. Aspergillus flavus, Candida krusei and Botrytis cinerea. Quantification by HPLC showed the content of Flindersine and Ulopterol to be 0.361% and 0.266% respectively on dry weight basis of the leaves. Conclusions: Ethyl acetate extract (successive extraction) contained Ulopterol, a coumarin, besides Flindersine, a quinolone alkaloid, as a major active principle in the antimicrobial studies. This is the first report of the antimicrobial activity of Ulopterol and also its first report from the plant. [Copyright &y& Elsevier]