Quantitative Analysis of Bioactive Carbazole Alkaloids in Murraya koenigii (L.) from Six Different Climatic Zones of India Using UPLC/MS/MS and Their Principal Component Analysis.

Murraya koenigii (L.) Spreng (Curry leaf) is a commercially important medicinal plant in South Asia, containing therapeutically valuable carbazole alkaloids (CAs). Thus, the quantitative evaluation of these compounds from different climatic zones of India are an important aspect for quality assessment and economic isolation of targeted compounds from the plant. In this study, quantitative estimation of CAs among 34 Indian natural populations of M. koenigii was assessed using UPLC/MS/MS. The collected populations represent the humid subtropical, tropical wet & dry, tropical wet, semi-arid, arid, and montane climatic zones of India. A total of 11 CAs viz. koenine-I, murrayamine A, koenigine, koenimbidine, koenimbine, O-methylmurrayamine A, girinimbine, mahanine, 8,8''-biskoenigine, isomahanimbine, and mahanimbine were quantified using multiple reaction monitoring (MRM) experiments within 5.0 min. The respective range for natural abundance of CAs were observed as 0.097-1.222, 0.092-5.014, 0.034-0.661, 0.010-1.673, 0.013-7.336, 0.010-0.310, 0.010-0.114, 0.049-5.288, 0.031-1.731, 0.491-3.791, and 0.492-5.399 mg/g in leaves of M. koenigii. The developed method shown linearity regression coefficient (r2 >0.9995), LOD (0.003-0.248 ng/mL), LOQ (0.009-0.754 ng/mL), and the recovery was between 88.803-103.729 %. The bulk of these CAs were recorded in their highest concentrations in the humid subtropical zone, followed by the tropical wet & dry zones of India. Further, principal component analysis (PCA) was performed which differentiated the climatic zones according to the dominant and significant CAs contents within the populations. The study concludes that the method established is simple, rapid, with high sample throughput, and can be used as a tool for commercial purposes and quality control of M. koenigii.

Phytochemical portfolio and anticancer activity of Murraya koenigii and its primary active component, mahanine.

Murraya koenigii, a plant belonging to the Rutaceae family is widely distributed in Eastern-Asia and its medicinal properties are well documented in Ayurveda, the traditional Indian system of medicine. Through systematic research and pharmacological evaluation of different parts of the plant extracts has been shown to possess antiviral, anti-inflammatory, antioxidant, antidiabetic, antidiarrhoeal, antileishmanial, and antitumor activity. In the plant extracts, carbazole alkaloid, mahanine has been identified as the principle bioactive component among several other chemical constituents. Scientific evidence derived not only from in vitro cellular experiments but also from in vivo studies in various cancer models is accumulating for the pronounced anticancer effects of mahanine. The primary objective of this review is to summarize research data on cytotoxic chemical constituents present in different parts of Murraya koenigii and the anticancer activity of mahanine along with the recent understanding on the mechanism of its action in diverse cancer models. The information on its bioavailability and the toxicity generated from the recent studies have also been incorporated in the review.

In vivo antianxiety and antidepressant activity of Murraya paniculata leaf extracts.

OBJECTIVE: Different parts of Murraya paniculata have been used traditionally for treating several ailments including mental disorders. The present study was designed to evaluate the antianxiety and antidepressant potential of M. paniculata leaves using elevated plus maze model and forced swim test, respectively. METHODS: Extracts of M. paniculata made with petroleum ether (60-80 °C), chloroform, ethanol and water were evaluated for antianxiety and antidepressant activity. The anxiolytic chloroform extract was subjected to column chromatography, yielding five fractions (F1-F5). Fraction F5 (100 mg/kg), which showed notable anxiolytic activity, was further chromatographed to get four subfractions (F5.1-F5.4). Simultaneously, the ethanol extract was partitioned with ethyl acetate to obtain ethyl acetate soluble fraction (EASF) and ethyl acetate insoluble fraction. Phytochemical screening of bioactive extracts/fractions and detection of mahanimbine in M. paniculata leaf extract by thin-layer chromatography was also carried out. RESULTS: Fraction F5.3 (25 mg/kg) and EASF (20 mg/kg) showed significant anxiolytic and antidepressant activity, respectively. Thin-layer chromatography and phytochemical screening demonstrated the absence of mahanimbine in M. paniculata leaves. Coumarins were observed to be responsible for the anxiolytic activity. CONCLUSION: The results validate the traditional use of M. paniculata leaves in the treatment of mental disorders.

New furocarbazole alkaloids from Lonicera quinquelocularis.

Two new furocarbazole alkaloids, 3-formyl-6,7-dimethoxy-furo[1,2]carbazole (1) and methyl-6,7-dimethoxy-furo[1,2]carbazole-3-carboxylate (2), along with two known carbazole alkaloids, 3-formyl-2-hydroxy-7-methoxycarbazole (3) and methyl 2,7-dimethoxycarbazole-3-carboxylate (4) were isolated from the ethyl acetate soluble fraction of Lonicera quinquelocularis. Their structures were established on the basis of spectroscopic analysis.

Identification and characterization of an anti-pseudomonal dichlorocarbazol derivative displaying anti-biofilm activity.

Pseudomonas aeruginosa strains resistant towards all currently available antibiotics are increasingly encountered, raising the need for new anti-pseudomonal drugs. We therefore conducted a medium-throughput screen of a small-molecule collection resulting in the identification of the N-alkylated 3,6-dihalogenocarbazol 1-(sec-butylamino)-3-(3,6-dichloro-9H-carbazol-9-yl)propan-2-ol (MIC = 18.5 µg mL⁻¹). This compound, compound 1, is bacteriostatic towards a broad spectrum of Gram-positive and Gram-negative pathogens, including P. aeruginosa. Importantly, 1 also eradicates mature biofilms of P. aeruginosa. 1 displays no cytotoxicity against various human cell types, pointing to its potential for further development as a novel antibacterial drug.

Development and thermodynamic evaluation of novel lipid raft stationary phase chromatography for screening potential antitumor agents.

Novel lipid raft stationary phase chromatography (LRSC), with lipid rafts that contain abundant tropomyosin-related tyrosine kinase A receptors immobilized on the stationary phase, was developed for a high-throughput screening of potentially active antitumor agents. Lestaurtinib was used as a model compound to determine the operational parameters of the LRSC. Of all the factors considered, the particle size of column packing, the column temperature and the flow rate were of immense importance in determining the performance of the established LRSC system. In order to profoundly comprehend the binding interaction between the model drug and the receptors on the column, thermodynamic studies were employed. The results revealed that the interaction was spontaneous and exothermic, a typical enthalpy-driven process. Additionally, the primary forces were hydrogen bonding and van der Waals forces. In evaluating the applicability of the method, active extracts from Albizziae Cortex were screened out using the LRSC system under the optimized conditions. The bioactive components were successfully confirmed by the MTT assay. In conclusion, it could be said that the LRSC is a good model for screening potential antitumor agents because of its viability, rapid response and scalable features.

RP-HPLC-DAD for simultaneous estimation of mahanine and mahanimbine in Murraya koenigii.

Murraya koenigii leaves (Rutaceae) are widely used as food condiments in various food preparations in India. They possess a wide range of biological activities including antioxidant, antibacterial, anticancer, hypoglycemic and hypolipidemic activity. A rapid reverse-phase high-performance liquid chromatography (RP-HPLC) method has been developed for quantitative estimation of mahanine and mahanimbine, two major bioactive alkaloids in this plant. The amounts of mahanine and mahanimbine were detected as 9.56 ± 1.04 and 4.32 ± 0.81% w/w in the extract, with the retention times of 6.26 ± 0.66 and 10.40 ± 0.95 minutes. The limits of detection and quantification were estimated to be 29.30 and 81.12 µg/mL and 1.67 and 6.31 µg/mL, respectively. This specific and precise validated method can be useful for the routine analysis and quantitative determination of mahanine and mahanimbine in this therapeutically potent medicinal plant.

Challenge in the speciation of nitrogen-containing compounds in heavy petroleum fractions by high temperature comprehensive two-dimensional gas chromatography.

Extending the knowledge related to nitrogen-containing compounds presents an important interest for the petroleum industry due to their implication in atmosphere pollution as well as their inhibitive or refractive behaviour towards hydroprocessing. Most of the nitrogenated species are concentrated in heavy petroleum cuts. As no analytical method is resolutive enough for these heavy cuts, particularly regarding nitrogen-containing compounds, a new approach is needed. For this reason, this study focuses on the development of a GC×GC technique, through the hyphenation of a specific NCD detector with a GC×GC system at high temperature. The performances of highly polar thermally stable stationary phases, in particular those composed of promising ionic liquids, were monitored in normal and reversed configurations. Subsequently, after the extraction of neutral or basic compounds by adsorption on an ion-exchange resin, a first quantitative determination was attempted for a straight-run and a direct coal liquefaction vacuum gas oils. This study led to a better understanding of the behaviour of highly aromatic N-compounds by 2D-GC including an ionic liquid phase as well as a deeper N-characterization of heavy petroleum fractions.

Hepatoprotective effects of aqueous leaf extract and crude isolates of Murraya koenigii against in vitro ethanol-induced hepatotoxicity model.

Medicinal plants constitute a principal health care resource corroborating their gradual acceptance by the global population. The ethno medicinal plant, Murraya koeniggi (Curry-leaf tree) as is native to India exhibits diverse biological activities. Unpublished data from our laboratory revealed hepatoprotective activity of its crude aqueous extract against ethanol-induced hepatotoxicity in experimental animals. Chronic ethanol consumption diminishes the cellular antioxidant levels through free radical induced injury causing hepatitis and cirrhosis with mortality in severe cases. This provided a rationale for studying its mechanistic approaches in terms of modulation of antioxidant defenses for probable hepatoprotective activity against ethanol-induced hepatotoxicity in vitro. Based on the inhibitory concentration (IC(50)) obtained from the cell viability assay, graded concentrations of 100 µg/ml and 500 µg/ml of aqueous extract (WE), isolated carbazole alkaloids (CA) and tannin (T) fraction were chosen to study the hepatoprotective activity against ethanol-induced hepatotoxicity using liver carcinoma cell lines (Hep G(2)). Their antioxidant activity with anti-lipid peroxidation potential (LPO), effects on protein content, liver metabolizing enzymes viz., glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and the morphology of the cells were studied as parameters of hepatoprotection. The tannins and the carbazole alkaloids from the aqueous extract exhibited excellent hepatoprotective activity with respect to the different parameters studied and maintained normal morphology even after ethanolic challenge to the cells as comparable to the protection offered by the standard drug L-ornithine L-aspartate (LOLA). The modulating effect of the aqueous extract and isolates on liver metabolizing enzymes, reduction in lipid peroxidation and decreased cellular damage were found to contribute to the hepatoprotective activity.

Determination of a pyranocoumarin and three carbazole compounds in Clausena excavata by RP-HPLC.

Clausenidin, O-methylmukonal, 3-formyl-2,7-dimethoxycarbazole, and clauszoline-J, isolated from the rhizomes and roots of Clausena excavata, exhibit anti-HIV-1 activity in a syncytial assay with EC(50) values of 5.3, 12.0, 29.1, and 34.2 microM, respectively. Due to the highly active anti-HIV-1 property, quantitative analysis of four compounds are investigated. The direct analysis of these four compounds in the crude extracts of the combined rhizomes and roots of Clausena excavata from ten various sources in Thailand by high-performance liquid chromatography is accomplished. Chromatographic separation is achieved on a C(18) column, and the mobile phase is a mixture of methanol and distilled water in a mode of isocratic or gradient elution detected at 254 nm at a flow rate of 0.6 mL/min for clausenidin, at 274 nm at a flow rate of 0.6 mL/min for O-methylmukonal, at 298 nm at a flow rate of 0.4 mL/min for 3-formyl-2,7-dimethoxycarbazole, and at 242 nm at a flow rate of 0.4 mL/min for clauszoline-J. This is the first quantitative analysis of these four anti-HIV-1 compounds from the crude extract without prior isolation and purification steps.

Murrayakoeninol--a new carbazole alkaloid from Murraya koenigii (Linn) Spreng.

A new carbazole alkaloid, designated as murrayakoeninol, was isolated from the leaves of Murraya koenigii (Linn) Spreng, along with four known carbazole alkaloids, viz. mahanimbine, koenimbine, O-methylmurrayamine-A and murrayazolinine and one from the bark viz. girinimbine. The structure of the new alkaloid 1 was elucidated on the basis of 2D NMR spectral analysis and chemical reactions.

Microcosm experiments of oil degradation by microbial mats.

Several microcosm experiments were run in parallel to evaluate the efficiency of microbial mats for crude oil degradation as compared with physico-chemical weathering. The oils used in the experiments constituted representative examples of those currently used for commercial purposes. One was aliphatic and of low viscosity (33.4 American Petroleum Institute degrees, degrees API) and the other was predominantly aromatic, with high sulphur content (ca. 2.7%) and viscosity (16.6 degrees API). After crude oil introduction, the microcosms were kept under cyclic changes in water level to mimic coastal tidal movements. The transformations observed showed that water weathering leads to more effective and rapid elimination of low molecular weight hydrocarbons than microbial mat metabolism, e.g. n-alkanes with chain length shorter than n-pentadecane or n-heptadecane, regular isoprenoid hydrocarbons with chain length lower than C16 or C18 or lower molecular weight naphthalenes. Microbial mats preserved these hydrocarbons from volatilization and water washing. However, hydrocarbons of lower volatility such as the C24-C30 n-alkanes or containing nitrogen atoms, e.g. carbazoles, were eliminated in higher proportion by microbial mats than by water weathering. The strong differences in composition between the two oils used for the experiments were also reflected in significant differences between water weathering and microbial mat biodegradation. Higher oil viscosity seemed to hinder the former but not the later.

Identification of alkyl carbazoles and alkyl benzocarbazoles in Brazilian petroleum derivatives.

Carbozoles are important compounds in crude oils, as they may be used as geochemical tracers, being the major type of nitrogen compounds in petroleum. At the same time, they are regarded as undesirable due to the problems they may cause in the refining process, such as catalyst poisoning, corrosion, gum or color formation in final products. As separation and identification of carbazoles are challenging goals, this work presents a chromatographic method, made of a pre-fractionation on neutral alumina followed by the separation and identification of two classes of carbazoles using FeCl(3)/Chromossorb W and gas chromatograph with mass spectrometer (GC/MS) (SIM-single ion monitoring mode) analysis. For the first time, a series of alkyl carbazoles and alkyl benzocarbazoles were identified in heavy gas oil (HGO) and atmospheric residue of distillation (ARD) obtained from Brazilian petroleum.

Correlation of systemic and developmental toxicities with chemical component classes of refinery streams.

Refinery streams are complex mixtures, but of a relatively few homologous series of hydrocarbons (paraffins, olefins, naphthenics, and aromatics). Studies were performed to determine if systemic and developmental toxicity were related to the presence and levels of certain classes of refinery stream components. We have performed systemic toxicology studies in the rat on 13 refinery streams: Clarified Slurry Oil, Coker Light Gas Oil, Distillate Aromatic Extract, Heavy Atmospheric Gas Oil, Heavy Coker Gas Oil (from three refineries), Heavy Vacuum Gas Oil, Light Catalytically Cracked Naphtha, Light Cycle Oil, Syntower Bottoms, Vacuum Tower Overhead, and Visbreaker Gas Oil. Rats were exposed via repeated dermal administration (daily) at several dose levels. Developmental toxicology studies were performed on these same streams with the following exceptions: only two Heavy Coker Gas Oils were tested and Visbreaker Gas Oil was not tested. End points for systemic toxicity (13-week) studies included skin irritation, body and organ weights, hematology, and serum chemistry; for developmental toxicity studies some of these same end points (excluding hematology) were considered, but they also included resorption and fetal body weight. In general, toxicity was correlated with concentrations of polycyclic aromatic compounds (PAC) composed of 3, 4, 5, 6, and/or 7 rings (decreased thymus weight, increased liver weight, aberrant hematology and serum chemistry, increased incidence of resorption, decreased fetal body weight), PAC containing nonbasic nitrogen heteroatoms (increased mortality, decreased body weight, decreased thymus weight, increased liver weight, decreased hemoglobin content and hematocrit level, decreased fetal body weight), and/or PAC containing sulfur heteroatoms (decreased red blood cell and platelet counts, increased sorbitol dehydrogenase.)(ABSTRACT TRUNCATED AT 250 WORDS)

Clausine-D and -F, two new 4-prenylcarbazole alkaloids from Clausena excavata.

Two new 4-prenylcarbazole alkaloids, clausine-D (1) and -F (3) were isolated from the stem bark of Clausena excavata. Their structures were elucidated by spectral methods. Both of clausine-D (1) and -F (3) showed significant antiplatelet aggregation activity.