Identification Of Active Ingredients Research Articles

Native Medicinal Plants and Their Effects on Anti-biofilm and Anti-quorum Sensing Activity

Background: Pseudomonas aeruginosa is an opportunistic pathogen with a high mortality rate, particularly among immunocompromised patients. The increase in antibiotic resistance in P. aeruginosa isolates from clinical samples has prompted researchers to seek alternative ways to reduce the pathogenicity of this microorganism. Targeting quorum sensing (QS) presents a promising approach for limiting infections caused by this bacterium. Objectives: This study aimed to evaluate mechanisms for reducing QS and controlling bacterial growth using various medicinal plants. Methods: In this study, the anti-QS effects of seven plant extracts—thyme (Thymus vulgaris), chamomile (Matricaria chamomilla), sumac (Rhus coriaria), ginger (Zingiber officinale), lemon balm (Melissa officinalis), rosehip (Echinacea angustifolia), and marshmallow (Althaea officinalis)—were investigated against P. aeruginosa. A total of 180 P. aeruginosa clinical isolates were examined. High-performance liquid chromatography (HPLC) was then used to detect phenolic compounds and flavonoids in the seven extracts. The presence of quorum-sensing genes (lasR, rhlR, and rhlI) was also evaluated. In addition, pyocyanin quantification, swarming motility assays, and biofilm formation assays were conducted with the herbal extracts. Results: The HPLC analysis results were determined based on relative times and peak areas, revealing specific peaks at different points. Molecular methods confirmed the presence of QS genes, including LasI (54%), lasR (34%), rhlR (12%), and rhlI (4%), which were used in anti-QS screening. Expression of the LasI gene decreased in P. aeruginosa strains treated with herbal extracts compared to untreated strains. Notably, Thymus vulgaris extract demonstrated the strongest inhibition, reducing violacein production by 70.5%, suppressing pyocyanin by 94%, and inhibiting swarming motility and biofilm formation by 80.3%. Conclusions: This study demonstrated that herbal extracts influence virulence factors and gene expression, particularly targeting the las gene, in P. aeruginosa isolates. We recommend further research on the extraction and identification of active ingredients, which may hold potential as therapeutic agents.

Identification of active ingredients in Naomaitai capsules using high-resolution mass spectrometry unite molecular network analysis and prediction of their action mechanisms.

Although Naomaitai capsule (NMC) is widely used in clinical practice and has a good curative effect for cerebral infarction, its material basis and mechanism of action remain unclear. In this study, ultra-high-performance liquid chromatography (UHPLC) coupled with quadrupole Orbitrap MS technology was used to analyse the in vivo and in vitro components of NMC, and the Global Natural Products Social Molecular Networking website was used to further analyse the components of NMC. Next, systems biology approaches were employed to investigate the mechanism of action of NMC. Finally, molecular docking technology was used to verify the network pharmacological results. In total, 177 compounds were identified in vitro, including 65 terpenoids, 62 flavonoids, 25 organic acids and 11 quinones. 64 compounds were identified in the blood of mice, and the main active components included ginkgolide C, ginkgolide A, ligustilide, tanshinone IIB, olmelin, emodin and puerarin. The main targets in vivo included TP53, SRC, STAT3, PIK3CA and PIK3R1. In conclusion, this study has revealed that NMC acts on multiple targets in the body through various active components, exerting synergistic effects in the treatment of CI. Its mechanism of action may involve inhibiting neuronal apoptosis, oxidative stress and inflammatory responses as well as reducing cerebral vascular permeability and promoting cerebral vascular regeneration.

Rapid Targeted Screening and Identification of Active Ingredients in Herbal Extracts through Ligand-Detected NMR and Database Matching.

Herbal extracts are rich sources of active compounds that can be used for drug screening due to their diverse and unique chemical structures. However, traditional methods for screening these compounds are notably laborious and time-consuming. In this manuscript, we introduce a new high-throughput approach that combines nuclear magnetic resonance (NMR) spectroscopy with a tailored database and algorithm to rapidly identify bioactive components in herbal extracts. This method distinguishes characteristic signals and structural motifs of active constituents in the raw extracts through a relaxation-weighted technique, particularly utilizing the perfect echo Carr-Purcell-Meiboom-Gill (peCPMG) sequence, complemented by precise 2D spectroscopic strategies. The cornerstone of our approach is a customized database designed to filter potential compounds based on defined parameters, such as the presence of CHn segments and unique chemical shifts, thereby expediting the identification of promising compounds. This innovative technique was applied to identifying substances interacting with choline kinase α (ChoKα1), resulting in the discovery of four new inhibitors. Our findings demonstrate a powerful tool for unraveling the complex chemical landscape of herbal extracts, considerably facilitating the search for new pharmaceutical candidates. This approach offers an efficient alternative to traditional methods in the quest for drug discovery from natural sources.

Research progress on Wuzi Yanzong pills in the treatment of male infertility

Abstract In recent years, because of changes in lifestyle, social environments, and other factors, male fertility has exhibited a declining trend in various countries and regions, including China. This poses a challenge for the prevention and treatment of male infertility. Wuzi Yanzong pill (WZYZP) is a renowned Chinese medicinal formula utilized for the treatment of male infertility and boasts a long history of use in this context. This article summarized the treatment approaches used in both Chinese and Western medicine for male infertility, along with the research advancements pertaining to the analysis of Chinese medical theory, pharmacological effects, bioactive ingredients, and the establishment of quality control standards for WZYZP in the treatment of male infertility, aiming to provide references for research related to the mechanism of action, identification of active ingredients, and clinical application of WZYZP in the treatment of male infertility.

Isolation and identification of active ingredients and biological activity of Dioscorea nipponica Makino

This study reported the isolation and identification of bioactive compounds from Dioscorea nipponica Makino, a plant used in traditional medicine for various ailments. Nine compounds were isolated, including a new compound named as diosniposide E, which was elucidated by analyzing its 1H-NMR, 13C-NMR, DEPT, COSY, HMBC and MS data and comparing them with data available in literature. The other eight compounds were identified as known compounds. Theoretical calculations of energy and the generation of a molecular electrostatic potential surface map were employed to assess the antioxidant capacity of nine compounds, the calculation results exhibited that compounds 5 and 6 had strong antioxidant capacities. To further evaluate the antioxidant activities of the investigated compounds, the DPPH and ABTS assays were conducted. The results from the DPPH scavenging activity test revealed that compounds 4–6 exhibited enhanced scavenging activities compared to L-ascorbic acid, while displaying similar efficacy to trolox. Moreover, the ABTS scavenging activities of compounds 4–6 were found to surpass those of L-ascorbic acid and trolox. In terms of α-glucosidase inhibition, compounds 3 and 4 displayed remarkable inhibitory activities that surpassed the effects of acarbose. Additionally, compound 2 exhibited potent anticholinesterase activities, outperforming donepezil. This research provides insights into the potential bioactive compounds present in Dioscoreanipponica Makino and may contribute to its use in traditional medicine.

Phytochemical Analysis of Acaciaehrenbergiana (Hayne) Grown in Qatar: Identification of Active Ingredients and Their Biological Activities.

Acacia ehrenbergiana (Hayne), also known as Salam, is a highly drought resistant shrub distributed in North and East Africa, and the Arabian Peninsula. The plant is gathered for its gum and fiber, and is an important legume species for indigenous populations. In this study, the phytochemical analysis, antibacterial, and antioxidant properties of various alcoholic and aqueous extracts of Acacia ehrenbergiana grown in Qatar were investigated. The qualitative phytochemical screening of this species exhibited the presence of glycosides, tannins, flavonoids, terpenoids, saponins, phenol, and anthraquinones in various extracts. The agar diffusion method was performed to check the antibacterial activity. The acetone and ethanol extracts showed 85% antibacterial activity of the control against Gram-negative E. coli, while the acetone extract had 65% activity against the Bacillus Gram-positive species. The highest activity against Staphylococcus aureus was 65% for the butanol extract. The antioxidant capacities were evaluated by the DPPH method. Various extracts exhibited antioxidant activities similar to or higher than standard antioxidants, with the highest percent inhibition of 95% for the acetone and ethanol extracts. The acetone extracts were further purified by reverse phase combiflash chromatography followed by HPLC. Three of the pure compounds isolated were subjected to MS, FTIR, and NMR spectral analysis and were found to be stigmasterol, spinasterol, and theogallin. In conclusion, the observed antibacterial and antioxidant activities as well as the presence of secondary metabolites with potential medicinal activities makes Acacia ehrenbergiana a potent valuable endemic medicinal plant.

Network pharmacology and molecular docking technology-based predictive study of the active ingredients and potential targets of rhubarb for the treatment of diabetic nephropathy

Diabetic nephropathy (DN) is one of the most serious complications of diabetes and the main cause of end-stage renal failure. Rhubarb is a widely used traditional Chinese herb, and it has exhibited efficacy in reducing proteinuria, lowering blood sugar levels and improving kidney function in patients with DN. However, the exact pharmacological mechanism by rhubarb improves DN remain unclear due to the complexity of its ingredients. Hence, we systematically explored the underlying mechanisms of rhubarb in the treatment of DN. We adopted a network pharmacology approach, focusing on the identification of active ingredients, drug target prediction, gene collection, Gene Ontology enrichment and Kyoto Encyclopedia of Genes and Genomes enrichment. Molecular docking technology was used to verify the binding ability between the main active compounds and central therapeutic targets, and screen out the core active ingredients in rhubarb for the treatment of DN. Finally, molecular dynamics simulation was performed for the optimal core protein-ligand obtained by molecular docking using GROMACS software. The network analysis identified 16 active compounds in rhubarb that were linked to 37 possible therapeutic targets related to DN. Through protein–protein interaction analysis, TP53, CASP8, CASP3, MYC, JUN and PTGS2 were identified as the key therapeutic targets. By validation of molecular docking, finding that the central therapeutic targets have good affinities with the main active compounds of rhubarb, and rhein, beta-sitosterol and aloe-emodin were identified as the core active ingredients in rhubarb for the treatment of DN. Results from molecular dynamics simulations showed that TP53 and aloe-emodin bound very stably with a binding free energy of − 26.98 kcal/mol between the two. The results of the gene enrichment analysis revealed that the PI3K-Akt signalling pathway, p53 signalling pathway, AGE-RAGE signalling pathway and MAPK signalling pathway might be the key pathways for the treatment of DN, and these pathways were involved in podocyte apoptosis, glomerular mesangial cell proliferation, inflammation and renal fibrosis. Based on the network pharmacology approach and molecular docking technology, we successfully predicted the active compounds and their respective targets. In addition, we illustrated the molecular mechanisms that mediate the therapeutic effects of rhubarb against DN. These findings provided an important scientific basis for further research of the mechanism of rhubarb in the treatment of DN.

Assessment of the Quality of Injectable Antibiotics in Benin.

Substandard and falsified medicines are an enormous threat to global health. Poor quality antibiotic preparations contribute to the development of antimicrobial resistance. In surgery, where the occurrence of healthcare-associated infections is high, healthcare teams need to rely on the quality of antibiotic prophylaxis to prevent infections. We assessed the quality of antibiotics used for surgical infection prophylaxis in Benin. Thirty-three samples were collected from six hospitals located in various departments in Benin. The antibiotics (powders for injection: amoxicillin + clavulanic acid, ampicillin, ceftriaxone; solutions for injection: ciprofloxacin, gentamicin, metronidazole) were assessed using visual inspection, pharmacotechnical tests (including uniformity of mass, pH measure, sterility test, and active pharmaceutical ingredient identification), and assay tests (including a simple analytical method thin layer chromatography) and complex analytical techniques (ultraviolet-visible spectrophotometry, high-performance liquid chromatography-diode-array detection, conductometry). Because the material needed for the methods recommended by the pharmacopeias to assess the dosage of gentamicin was not available, we developed and validated a conductometry method. Results showed that 97% (n = 32) of the samples passed visual inspection; 100% (n = 33) of the samples passed the pharmacotechnical tests, identification of active ingredients, and sterility test; 88% (n = 29) passed the test for percentage of active pharmaceutical ingredients. Overall, 15% of the samples did not pass the quality test (3% on visual inspection and 12% for excess active ingredients). Although most of the samples passed the quality tests, it appears important to perform routine quality control for intravenous medicines.

Integrative Analysis of the Transcriptome and Metabolome Reveals Genes Involved in Phenylpropanoid and Flavonoid Biosynthesis in the Trapa bispinosa Roxb.

BackgroundTrapa bispinosa Roxb. is grown worldwide as an important aquatic cash crop. Current research on Trapa bispinosa primarily focuses on the separation and identification of active ingredients, as well as the inhibitory effect on tumors; however, research on the molecular mechanism of secondary metabolite accumulation is rather limited. Consequently, an integrative analysis of transcriptome and metabolome is required to identify the key metabolic pathways, and key genes, and to explain the molecular mechanism of Trapa bispinosa.ResultsThe biosynthesis pathways of phenolics in Trapa bispinosa were examined through transcriptome and metabolome analyses. Transcriptome analysis yielded 42.76 million clean reads representing 81,417 unigenes with an average length of 1,752 bp. KEGG pathway analysis revealed that 1,623 unigenes, including 88 candidate unigenes related to phenolics biosynthesis, were up-regulated in Trapa bispinosa shell (FR) when compared to leaves (LF), root (RT), and stem (ST). The FR vs. LF group had the highest number of specific genes involved in phenylpropanoid, flavonoid, flavone, and flavonol biosynthesis pathways compared to all other comparison groups. In addition, RNA sequencing revealed 18,709 SSRs spanning 14,820 unigenes and 4,387 unigenes encoding transcription factors. Metabolome analysis identified 793 metabolites, including 136 flavonoids and 31 phenylpropane compounds. In the FR group compared to the LF group, there were 202 differentially accumulated metabolites (DAMs). The combined transcriptome and metabolome analyses indicated a significant correlation between 1,050 differentially expressed genes (DEGs) and 62 DAMs. This view proposes a schematic of flavonoid biosynthesis in the FR vs. LF group, providing evidence for the differences in genes and metabolites between FR and LF.ConclusionIn this study, through de novo transcriptome assembly and metabolome analysis, several DEGs and DAMs were identified, which were subsequently used to build flavonoid biosynthesis pathways and a correlation network. The findings pave the way for future research into the molecular mechanisms and functional characterization of Trapa bispinosa candidate genes for phenolics biosynthesis.

High-content phenotypic screening identifies novel chemistries that disrupt mosquito activity and development

New classes of chemistries are needed to control insecticide resistant populations of mosquitoes and prevent transmission of vector-borne diseases (VBDs). Organismal screens of chemical collections have played an important role in the search for new vector insecticides and the identification of active ingredients (AIs) that cause rapid mortality of mosquitoes. Advances in image-based screening offer an opportunity to identify chemistries that operate via novel biochemical modes and investigate the range of phenotypes exhibited by mosquitoes following exposure to lethal and sub-lethal chemical dose. An automated, high throughput phenotypic screen (HTS) employing high-content imaging of first instar (L1) Aedes aegypti larvae was developed to identify chemistries associated with mortality and atypical morphological phenotypes. A pilot screen of the Library of Pharmacologically Active Compounds (LOPAC1280) identified 92 chemistries that disrupted larval activity and development, including conventional insecticides and chemistries known to modulate G protein-coupled receptors (GPCRs) and other molecular targets in mammalian systems. Secondary assay series were used to evaluate a selection of chemistries for impacts on mosquito activity, survival and development. Ritodrine hydrochloride reduced mobility of larvae but had no observable effect on survival and development of mosquitoes. High doses of metergoline suppressed larval activity and sub-lethal dose resulted in pupal mortality. Assay data support the utility of phenotypic screening and diverse entomological end-points for discovery of novel insecticidal chemical scaffolds. The insecticide discovery process must consider how multi-modal efficacy spectra contribute to vector and VBD control.

Screening for Aldose Reductase Inhibitory Activity of 92 Crude Drugs, and Isolation and Identification of Active Ingredients from Clove

Screening for Aldose Reductase Inhibitory Activity of 92 Crude Drugs, and Isolation and Identification of Active Ingredients from Clove

Characterization of natural herbal medicines by thin-layer chromatography combined with laser ablation-assisted direct analysis in real-time mass spectrometry

The characterization and quality control of natural herbal medicines, such as traditional Chinese medicines (TCMs), is of great significance to ensure their potential efficacy and avoid severe side effects. Thin-layer chromatography (TLC) is a simple and classic approach for examining quality marker of natural products. Nevertheless, it is more difficult to further characterize the compounds adsorbed on the TLC plate. Herein, we reported a simple setup of laser ablation-assisted direct analysis in real-time mass spectrometry (LA-DART-MS), in which the coupling of mass spectrometry information to provide a predominant dimension in the identification of unknown chemical compositions separated on standard TLC plates, and it was applied for rapid characterization of various kinds of natural herbal medicines. The results showed that the introduction of low-cost small laser pointer had significantly improved the desorption process. The system was successfully applied in the analysis of alkaloids, flavonoids, anthraquinones, volatile oils, glycosides, organic acids, and eight different TCMs including Sophorae Flavescentis Radix, Angelicae Sinensis Radix, Acori Tatarinowii Rhizoma, Phellodendri Chinensis Cortex, Picrasmae Ramulus et Folium, Gynura Japonica, Rhei Radix et Rhizoma and Dendrobii Caulis. The obtained limits of detection (LODs) of this method for various types of reference substances were in the range of 4.6–162.2 ng/band on TLC plates. Furthermore, the quality control and identification of different species of Dendrobii Caulis herb was achieved. This study combines the advantages of TLC and ambient mass spectrometry to provide a good choice for the screening and identification of active ingredients and the quality evaluation of botanical samples.

Predictive Study of the Active Ingredients and Potential Targets of Codonopsis pilosula for the Treatment of Osteosarcoma via Network Pharmacology.

Osteosarcoma (OS) is the most common type of primary bone tumor in children and adults. Dangshen (Codonopsis pilosula) is a traditional Chinese medicine commonly used in the treatment of OS worldwide. However, the molecular mechanisms of Dangshen in OS remain unclear. Hence, in this study, we aimed to systematically explore the underlying mechanisms of Dangshen in the treatment of OS. Our study adopted a network pharmacology approach, focusing on the identification of active ingredients, drug target prediction, gene collection, gene ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and other network tools. The network analysis identified 15 active compounds in Dangshen that were linked to 48 possible therapeutic targets related to OS. The results of the gene enrichment analysis show that Dangshen produces a therapeutic effect in OS likely by regulating multiple pathways associated with DNA damage, cell proliferation, apoptosis, invasion, and migration. Based on the network pharmacology approach, we successfully predicted the active compounds and their respective targets. In addition, we illustrated the molecular mechanisms that mediate the therapeutic effect of Dangshen in OS. These findings may aid in the development of novel targeted therapies for OS in the future.

Pharmacognostic, Physicochemical and Anti-Oxidant Screening of Solenostemma Argel’s Leaf Extract

The present work deals with the identification of new approach towards the lead generation in medicinal chemistry and authenticating the reports present on the use of this Sudanese medicinal plant Solenostemma argel in the tribal region of Sudan. The pharmacognostic screening showed its physical and chemical composition with identification of active ingredients. The antioxidant activity was carried out by DPPH and nitric oxide scavenging method. In which ethanolic leaf extract showed prominent response which may be due to the promising presence of flavonoids and related poly phenolic compounds in it

Influence of different viscosity grade Methocel and Ethocel polymers for the development of controlled release dimenhydrinate matrix tablets

Dimenhydrinate is a therapeutic agent used for the treatment of nausea and vomiting. A conventional dose of dimenhydrinate is 50 mg per 8 h. Once-daily controlled release tablets of dimenhydrinate were formulated intended to treat emesis. The tablets were formulated using hydrophilic polymers Methocel® and hydrophobic polymers Ethocel® and subsequently analyzed for pre-compression and post-compression characteristics, % moisture uptake, dissolution profiles and release behavior of the drug. Identification of active ingredient and compatibility of blends were analyzed through FT-IR. Drug release kinetics of dimenhydrinate was assessed at pH 1.2, 4.5 and 6.8. The results of release profiles were evaluated for different kinetic models including model dependent and model independent. The optimized formulation K100M 30% showed ideal pre-compression properties and in vitro characteristics. Tablets exhibited a controlled drug release pattern due to the formation of a viscous gel layer followed by erosion. Furthermore, zero-order best described the release pattern with the value of r2 ˃ 0.99. Once-daily tablets released 98–100% drug in 24 h. Significantly more concentration and higher viscosity grade of hydrophilic polymer ensured the required pattern of drug release.

Effects of Methanol Extract of Illicium verum Hook.f on Coagulation Parameters in Rabbits

Background: Chinese Star anise is known as Illicium verum hook.f; a distinguished spice utilized in dishes, especially in the Asian region. However, it was not evaluated for its anticoagulant potential. The study was designed specifically to evaluate the effect of methanol extract of Illicium verum hook.f on various coagulation parameters in animal models. Methods: In the study, effect of methanol extract of I. verum hook.f was evaluated on various coagulation parameters such as Prothrombin time (PT), activated partial thromboplastin time (aPTT) and thrombin time (TT) in rabbits at three different doses, i.e. 150, 250 and 350 mg/kg body weight after 60 days of continuous dosing. Comparison was also done with the standard anticoagulant drug i.e., Warfarin. Data was compared with control group’s values by One-Way ANOVA trailed by post hoc Tukey’s test. Results: The current analysis revealed that administration of methanol extract of I. verum hook.f at 250 mg of dose for 30 days resulted in elevation of PT, aPTT and TT, whereas, highly significant increase was observed in PT and aPTT at the 350 mg/kg. However, administration of the extract for 60 days at 250 mg/ kg resulted in significant increase in PT while, significant increase in aPTT. Administration at 350 mg/kg of dose for same period resulted in significant rise in PT and aPTT in contrast to TT. Conclusion: Methanol’s extract of I. verum hook.f (star anise) showed significant anticoagulant effects, which is important in hypercoagulable conditions and cardiovascular diseases. However, isolation and identification of active ingredients is required to further validate the results.

Antimalarial herbal remedies of Bukavu and Uvira areas in DR Congo: An ethnobotanical survey.

The main objective of the present study was to collect and gather information on herbal remedies traditionally used for the treatment of malaria in Bukavu and Uvira, two towns of the South Kivu province in DRC. Direct interview with field enquiries allowed collecting ethnobotanical data; for each plant, a specimen was harvested in the presence of the interviewed traditional healers (THs). The recorded information included vernacular names, morphological parts of plants, methods of preparation and administration of remedies, dosage and treatment duration. Plants were identified with the help of botanists in the herbaria of INERA/KIPOPO (DRC) and the Botanic Garden of Meise (Belgium), where voucher specimens have been deposited. The results were analysed and discussed in the context of previous published data. Interviewees cited 45 plant species belonging to 41 genera and 21 families used for the treatment of malaria. These plants are used in the preparation of 52 recipes, including 25 multi-herbal recipes and 27 mono-herbal recipes. Apart of Artemisia annua L. (Asteraceae; % Citation frequency=34%) and Carica papaya L. (Caricaceae; % Citation frequency=34%), the study has highlighted that the most represented families are Asteraceae with 12 species (26%), followed by Fabaceae with 7 species (16%) and Rubiaceae with 4 species (9%). For a majority of plants, herbal medicines are prepared from the leaves in the form of decoction and administered by oral route. Literature data indicate that part of cited species are already known (38%) and/or studied (30%) for antimalarial properties, which gives credit to the experience of Bukavu and Uvira interviewees and some level of confidence on collected information. The highly cited plants should be investigated in details for the isolation and identification of active ingredients, a contribution to the discovery of new possibly effective antimalarials.

Strategy for rapid screening of antioxidant and anti-inflammatory active ingredients in Gynura procumbens (Lour.) Merr. based on UHPLC-Q-TOF-MS/MS and characteristic ion filtration.

Gynura procumbens (Lour.) Merr. is traditionally used as a raw material for making dumplings or steamed stuffed buns, and its fresh leaves are boiled with water for tea. Herein, we established an ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS/MS) combined with characteristic ion filtration (CIF) strategy to rapidly screen active ingredients with antioxidant and anti-inflammatory properties in G. procumbens. This strategy involved screening the active part of G. procumbens using antioxidation and anti-inflammatory activity assays; discovering the active compounds by speculating on the active site's chemical composition by UHPLC-Q-TOF-MS/MS plus CIF; and verifying the active compounds' activities. The ethyl acetate extract (EEAF) of G. procumbens was the major active site. Eighty-one compounds were identified from the EEAF using UHPLC-Q-TOF-MS/MS plus CIF. Furthermore, polyphenols such as cynarine, isochlorogenic acids A and isochlorogenic acids C have excellent antioxidizing and anti-inflammatory activities. This study provides a practical strategy for rapid in vitro screening of the antioxidizing and anti-inflammatory activities of traditional vegetables and herbs and identification of active ingredients.

Antitumor activity of an Artemisia annua herbal preparation and identification of active ingredients

BackgroundArtemisia annua L. has gained increasing attention for its anticancer activity. However, beside artemisinin, less is known about the possible bioactive ingredients of Artemisia annua and respective herbal preparations. We hypothesized that, in addition to artemisinin, Artemisia annua preparations might contain multiple ingredients with potential anticancer activity. MethodsMDA-MB-231 triple negative human breast cancer (TNBC) cells along with other treatment resistant, metastatic cancer cell lines were used to investigate in vitro and in vivo the anticancer efficacy of an Artemisia annua extract marketed as a herbal preparation, which contained no detectable artemisinin (limit of detection = 0.2 ng/mg). The extract was characterized by HPLC-DAD and the most abundant compounds were identified by 1H- and 13C NMR spectroscopy and quantified by UHPLC-MS/MS. Cell viability and various apoptotic parameters were quantified by flow cytometry. In vitro data were validated in two in vivo cancer models, the chick chorioallantoic membrane (CAM) assay and in orthotopic breast cancer xenografts in nude mice. ResultsThe Artemisia annua extract, the activity of which could be enhanced by acetonitrile maceration, inhibited the viability of breast (MDA-MB-231 and MCF-7), pancreas (MIA PaCa-2), prostate (PC-3), non-small cell lung cancer (A459) cells, whereas normal mammary epithelial cells, lymphocytes, and PBMC were relatively resistant to extract treatment. Likewise, the extract's most abundant ingredients, chrysosplenol D, arteannuin B, and casticin, but not arteannuic acid or 6,7-dimethoxycoumarin, inhibited the viability of MDA-MB-231 breast cancer cells. The extract induced accumulation of multinucleated cancer cells within 24 h of treatment, increased the number of cells in the S and G2/M phases of the cell cycle, followed by loss of mitochondrial membrane potential, caspase 3 activation, and formation of an apoptotic hypodiploid cell population. Further, the extract inhibited cancer cell proliferation, decreased tumor growth, and induced apoptosis in vivo in TNBC MDA-MB-231 xenografts grown on CAM as well as in nude mice. ConclusionAn extract of an artemisinin-deficient Artemisia annua herbal preparation exhibits potent anticancer activity against triple negative human breast cancer. New active ingredients of Artemisia annua extract with potential anticancer activity have been identified.

Study of antispasmodic action of Lavandula angustifolia Mill hydroalcoholic extract on rat ileum

Introduction: Lavender (Lavandula angustifolia Mill) is a herbal medicine widely used for gastrointestinal (GI) disorders. However, its pharmacological action on isolated ileum has not been studied. In this research effect of hydroalcoholic extract of L. angustifolia on isolate ileum contractions was studies and compared with loperamide.Methods: Hydroalcoholic extract of the plant was prepared by percolation method. The total flavonoid contents were assessed by colorimetric technique. A portion of rat ileum was suspended in an organ bath containing Tyrode’s solution. The tissue was kept under 1 g tension at 37°C and continuously gassed with O2. The tissue was stimulated with KCl (80 mM), acetylcholine (ACh, 2 μM) and electrical field stimulation (EFS). Effect of the L. angustifolia extract was studied on ileum contractions and compared with that of loperamide.Results: The yield of hydroalcoholic extract was 17% with total flavonoid content of 185 μg/mL in the stock solution. Loperamide in concentration dependent manner inhibited ileum contractile response to KCl, ACh and EFS. Hydroalcoholic extract of L. angustifolia (8-512 μg/mL) concentration dependently inhibited ileum contraction induced by KCl (IC50 = 88 ± 21 μg/mL), ACh (IC50 = 119 ± 251 μg/mL) and EFS (IC50 = 87 ± 33 μg/mL). The vehicle had no significant effect on ileum contractions.Conclusion: From this study it was concluded that L. angustifolia extract at microgram concentration shows an inhibitory effect on rat ileum smooth muscle. Therefore, isolation and identification of active ingredients are recommended.