ISOLATION AND DETERMINATION CARPAINE ALKALOID IN PAPAYA (CARICA PAPAYA L.) LEAF EXTRACT BY THIN-LAYER CHROMATOGRAPHY SCANNER

Objective: This research was conducted to isolate the alkaloid carpaine by chromatography method and to determine it quantitatively by Thin Layer Chromatography Scanner. Methods: Dried leaves were macerated with ethanol 70% and fractionated with dichloromethane. Isolation of carpaine alkaloid from the dichloromethane fraction was carried out by column chromatography and preparative thin-layer chromatography according to the Rf value in Thin Layer Chromatography (TLC) after exposure by Dragendorff reagent. Results: The content of carpaine alkaloid was 7.5 mg with Rf 0.58 and dichloromethane: methanol (9.2:0.8) as eluent. Validation showed the linearity (R2) 0.9988, the limit of detection(LOD) was 0.05 ppm, the Limit of Quantification (LOQ) was 0.19 ppm, the recovery from 98.93-102.43%, and the % coefficient of variation was 0.16%. Conclusion: Carpaine alkaloid in papaya leaf extract was 10.52%.

Giving preparative thin layer chromatography some tender loving care

Preparative thin layer chromatography (prepTLC) is a commonly used method of purification suitable for small scale reactions. However, descriptions of the preferred methodology to load, run, and recover samples from prepTLC are non-standard and varied, making it part of the “hidden curriculum” of laboratory technique. In this article we report on the simple, cost-effective methods we use to load and collect samples from a plate, which enhance the convenience, speed, and precision of this technique.

In Vitro Anti-Trypanosoma cruzi Activity of Halophytes from Southern Portugal Reloaded: A Special Focus on Sea Fennel (Crithmum maritimum L.)

Marine halophytes are an outstanding reservoir of natural products and several species have anti-infectious traditional uses. However, reports about their potential use against neglected tropical ailments, such as Chagas disease, are scarce. This work evaluated for the first time the in vitro anti-Trypanosoma cruzi activity of extracts from the aromatic and medicinal species Helichrysum italicum subsp. picardii (Boiss. & Reut.) Franco (Asteraceae, everlasting) and Crithmum maritimum L. (Apiaceae, sea fennel). For that purpose, decoctions, tinctures, and essential oils from everlasting’s flowers and sea fennel’s stems, leaves, and flowers were tested against intracellular amastigotes of two T. cruzi strains. The extract from the sea fennel flower decoction displayed significant anti-trypanosomal activity and no toxicity towards the host cell (EC50 = 17.7 µg/mL, selectivity index > 5.65). Subsequent fractionation of this extract afforded 5 fractions that were re-tested in the same model of anti-parasitic activity. Fraction 1 was the most active and selective (EC50 = 0.47 μg/mL, selectivity index = 59.6) and was submitted to preparative thin-layer chromatography. One major compound was identified, falcarindiol, which was likely the one responsible for the observed anti-trypanosomal activity. This was confirmed using a commercially sourced molecule. Target-fishing studies showed falcarindiol as a ligand of T. cruzi spermidine synthase, pointing to a potential enzyme-inhibiting anti-trypanosomal mechanism of action. Overall, this work shows that sea fennel can provide effective anti-parasitic molecule(s) with potential pharmacological applications in the treatment of CD.

In Vivo Antimalarial Activity of Leaf Extracts and a Major Compound Isolated from Ranunculus multifidus Forsk

The leaves of Ranunculus multifidus Forsk. are traditionally used for the treatment of malaria in several African countries. In the present study, 80% methanol (RM-M) and hydrodistilled (RM-H) extracts of fresh leaves from R. multifidus and its major constituent anemonin were tested for their in vivo antimalarial activity against Plasmodium berghei in mice. Anemonin was also tested for its in vitro antimycobacterial activity against Mycobacterium smegmatis and M. abscessus in a microbroth dilution assay, and bacterial growth was analyzed by OD measurement. The isolation of anemonin from RM-H was carried out using preparative thin layer chromatography (PTLC). The chemical structures of anemonin and its hydrolysis product were elucidated using spectroscopic methods (HR–MS; 1D and 2D-NMR). Results of the study revealed that both RM-M and RM-H were active against P. berghei in mice, although the latter demonstrated superior activity (p < 0.001), as compared to the former. At a dose of 35.00 mg/kg/day, RM-H demonstrated a chemosuppression value of 70% in a 4-day suppressive test. In a 4-day suppressive, Rane’s and prophylactic antimalarial tests, anemonin showed median effective doses (ED50s) of 2.17, 2.78 and 2.70 μM, respectively. However, anemonin did not inhibit the growth of M. smegmatis and M. abscessus.

Isolation of the antidiarrhoeal tiliroside and its derivative from Waltheria indica leaf extract

The antidiarrhoeal effect of Waltheria indica methanol extract and fractions have been reported earlier but, the present work examined the intestinal relaxant effects of two flavonoid-phenyl propanoids isolated from the methanol extract. The active aqueous fraction was subjected to vacuum liquid chromatography using dichloromethane with increasing concentration of ethyl acetate, and that of methanol and water successively. The ten (10) fractions obtained were combined to give seven (7). The fraction 2 (C, D) was subjected to preparative thin layer chromatography on silica gel GF254 (10-40μm) using CHCl3-CH3OH (8:2) to obtain compound coded F2. Fraction 4 (F) was subjected to column chromatography using silica gel (60-120μm mesh) and eluted with dichloromethane with increasing concentrations of methanol. Fractions 9-28 were combined and subjected to column chromatography using chloroform with increasing concentration of methanol. The fractions 1-16 of these were purified on Sephadex LH-20 to obtain compound BAA. The identities of the two compounds were established using spectroscopic methods. The antidiarrheal effect of compound F2 was evaluated on mice using charcoal transit (100,200, 400mg/kg), castor oil (40, 60 mg/kg) while the two compounds were examined for their inhibitory effects on Ach-induced ileum contraction. The effects of the compounds were compared with loperamide (3mg/kg) and atropine (80μg). Compounds F2 and BAA were identified as tiliroside and 3’’’, 5’’’-dimethoxy tiliroside respectively. Tiliroside inhibited the charcoal transition in the animals in a dose dependent pattern with 400mg/ mL eliciting 63.41% inhibition compared to 59.23% produced by loperamide. The compound also elicited significantly (P<0.05) prolonged onset of stooling and reduced the number and weight of stools produced lower than the control. The two compounds drastically inhibited the Ach-induced contractions of the ileum. The compound, tiliroside at 10mg, completely abolished the contraction by Ach unlike 3’’’, 5’’’-dimethoxy tiliroside which reduced the contraction to 1.92% at 20mg. The identified compounds seem to be responsible for the ethnomedicinal use of the plant in treating diarrhea.

A Sesquiterpene Aldehyde Isolated From Ethyl Acetate Extract of Lansium Domesticum Fruit Peel

Lansium domesticum (fam. Meliaceae) contains various compounds with various biological activities. Based on the previous research, extracts from several parts of the plant have biological activity. This study aimed to isolate a compound from the fruitpeel of L. domesticum and evaluate cytotoxic activity against T47D, WiDr and HepG2 cell lines. Powdered peels were macerated with ethyl acetate and the filtrate was evaporated to give EtOAc extract. Dried extract was triturated with n-hexane to give n-hexane soluble fraction (A) and insoluble fraction (B). The fraction B was separated using vacuum column chromatography (VLC) with mobile phase n-hexane: ethyl acetate and given 5 fractions. Fractions B3-B5 were combined and separated using VLC with n-hexane and ethyl acetate as mobile phase. This VLC separation gave 18 subractions, subfractions 6-9 with the similar TLC profile were combined. This subfraction was separated further using preparative thin layer chromatography to give compound 1. The Isolated compound (1) appeared as liquid. The chemical structure of 1 was identified acoording to spectroscopic data and comparison with literature. Cytotoxic bioassay was performed on T-47D, WiDr and Hep G2 cell lines in a series of concentrations at 50, 40, 30, 20, 10 and 5µg/mL, with Doxorubicine used as positive control. According to spectroscopic data, compound 1 was identified as 2-ethyl,3-(1’-hydroxy-2’-menthene) propenal, and demonstrate the strongest cytotoxicity against T-47D cell lines (IC50=39.18+1.54 µg/mL).

Β-Sitosterol of Red Dragon Fruit (Hylocereus Polyrhizus) and Its Response to Macrophage And Nitric Oxide

Hylocerius polyrhizus has relatively big potency as natural antioxidant. The compound considered as antioxidant also has immunomodulatory activity. This study showed isolation for identifying the active compounds in H. polyrhizus peels that are able to increase immune system of human body. The methanol extracts were partitioned and fractionated. The active compounds of petroleum ether fraction were partitioned and purified using Preparative Thin Layer Chromatography (PTLC). The identification of active compound structures was done with spectroscopy: UV, FT-IR, 13CNMR, 1HNMR, DEPT and HSQC. The immunomodulatory activity was also tested. Based on the spectroscopic data, the identified isolate was β-sitosterol. The statistical analysis of macrophage cell activity and nitric oxide showed that isolates at the highest concentrations of 100 µg/mL were able to activate macrophage cells and enhance the production of nitric oxide.

Alkaloid profiling and antimicrobial activities of Papaver glaucum and P. decaisnei

Objective Papaver decaisnei Hochst. & Steud. Ex Elkan and Papaver glaucum Boiss. & Hausskn. growing wild in Northern Iraq have been historically used for medicinal purposes. In this study, both species were evaluated for their alkaloid content and antimicrobial activities. Results Alkaloids were extracted and isolated by preparative thin-layer chromatography (TLC). Identification was carried out by comparing spectral data (UV and 1H-NMR) and TLC Rf values with those of authentic samples. Two alkaloids, proapaorphine-type mecambrine and aporphine-type roemerine were isolated from P. decaisnei. Two benzylisoquinoline type alkaloids papaverine (major alkaloid) and palaudine as well as aporphine-type N-methylasimilobine have been obtained in P. glaucum. Both P. glaucum and P. decaisnei extracts revealed strong antimicrobial activity on Pseudomonas aeruginosa ATCC 27853 and Enterococcus faecalis ATCC 29212. Collectively these results indicate that P. glaucum and P. decaisnei are promising sources of alkaloids that could further be investigated for medicinal purposes.

Efficacy Of C-10 Massoialactone against-Multispecies Microbial Biofilm

Biofilm is a community of Microbes found in almost all habitats. They bind to the surface and become incorporated in the biomolecule-rich extracellular matrix. The matrix can fully Microorganisms are covered and rendered more immune to antibacterial agents, becoming a progressive infection source. Anti-biofilm compounds need to be discovered to deal with these biofilm-mediated infections. C-10 Massoialactone(C10H16O2), a significant component of MassoiaaromaticaBecc. The essential oil has demonstrated that antibacterial and anti-fungal properties are possible. This study aims to see how effective C-10 Massoialactone is against multi-species microbial biofilm of Candida albicans, Pseudomonas aeruginosa, and Escherichia coli. Massoia lactone It is derived from the essential oil of Massoiaaromatica bark. To isolate and explain these compounds, researchers used preparative thin layer chromatography (TLC), gas chromatography-mass spectrometry (GC-MS), with 1D-1H nuclear magnetic resonance (NMR) analysis. The impact of horse-tail lactone on several microbial biofilms was investigated. The biofilm structure was analyzed using a transmission electron microscope (TEM) and scanning electron microscope (SEM) the biofilm structure. GCMS and NMR results revealed the presence of C-10 Massoialactone (96.59 %). C-10 Massoialactoneshowed a dose-dependent activity in inhibiting biofilm formation as well as in breaking down established biofilms. Higher concentrations of C-10 Massoialactoneare required to inhibit the mature phase of the tested biofilm. TEM and SEM analysis showed apparent cell lysis in the biofilm in the presence of C-10 Massoialactone. C-10 Massoialactone can function because of its rich potential as a new anti-biofilm drug for various microbial biofilms.