Fraction 3 Research Articles

Bioassay-guided fractionation and in vitro and in silico biological activities of 2,4-di-tert-butylphenol isolated from Nocardiopsis sp. strain LC-8

In this study, an actinomycete strain was isolated from freshwater soil sediments to produce bioactive metabolite with a broad array of biological activities. Initially, preliminary antibacterial screening of isolate was performed by perpendicular streak method which showed the growth inhibition of all tested bacterial pathogens. Based on the culture characteristics, morphological identification, and 16S rRNA gene sequencing, the isolate was identified as Nocardiopsis sp. strain LC-8. Ethyl acetate extract of strain LC-8 showed potential antibacterial activity against tested bacterial pathogens with a maximum zone of inhibition of 16.98±0.57 mm against Staphylococcus aureus. After column chromatography, fraction 2 (F2) of the extract exhibited prominent growth inhibition activity against all tested bacterial and fungal pathogens. Antioxidant activity of F2 showed potent free radical scavenging activity with low IC50 values (DPPH - 278.15±0.35 μg/mL, ABTS - 367.55±1.13 μg/mL, and FRAP - 347.48±1.35 μg/mL). Total phenolic content of F2 was quantified as 362.44±1.25 mg gallic acid equivalent/gram extract. As per spectral analyses, the isolated compound from F2 was identified as 2,4-di-tert-butylphenol which further induced cytotoxic (IC50 value - 17.5±1.5 μg/mL) and apoptosis activity against MCF-7 cells. Molecular interaction studies of 2,4-di-tert-butylphenol with target proteins of pathogens and cancer cells displayed good docking score values. Finally, molecular dynamics simulation was performed to determine the stable structure of the protein-ligand system and effectiveness of 2,4-di-tert-butylphenol against the target protein. In conclusion, Nocardiopsis sp. strain LC-8 may act as a promising source for the production of 2,4-di-tert-butylphenol aiding diverse therapeutic roles.

Cytotoxic of Usnic Acid Isolated from Ramalina sp.

Ramalina sp. (Ramalinaceae) is a type of lichen known to contain many active secondary metabolite compounds that have the potential to be used as medicine or medicinal raw materials. One of the biological activities possessed by Ramalina sp. lichen is its anticancer activity. This research aims to isolate and identify active secondary metabolite compounds from the methanol extract of the Ramalina sp. lichen and to find out the cytotoxic activity of the isolated compound against MCF7 breast cancer cells. Compound 1 (usnic acid) was successfully isolated from fraction A. The isolation and purification process was carried out starting with a maceration process using acetone solvent, followed by silica gel column chromatography using a gradient solvent system consisting of n-hexane, n-hexane: EtOAc, EtOAc, EtOAc: MeOH, and MeOH with 5% increment of polarity to obtain 17 fractions (F-1 to F-17). From the 17 fractions obtained, fraction 3 (F-3) and fraction 4 (F-4) (eluted with n-hexane: EtOAc 30%), which had the same TLC profile, were combined and named as fraction A. Compound 1 (50 mg) is a yellow needle crystal that was formed in a bottle of fraction A, which was obtained after the process of combining fractions F-3 and F-4 and solvent evaporation process. The crystals were then separated and purified with CHCl3 and MeOH. Compound 1 was then characterized based on spectroscopic data. Various spectroscopic analysis data, including FTIR, 1D- and 2D-NMR, and LC-ESI-MS, show that Compound 1 is a dibenzofuran derivative compound with 18 carbons (3 from carbonyl groups (C=O) and 3 from methyl groups) and 2 hydroxyls (-OH). Cytotoxicity assay showed that at a low concentration of 18.75 ug/mL, Com-pound 1 caused a 67.06% decrease in MCF7 viability.

Identification and Molecular Docking Analysis of Angiotensin-Converting Enzyme Inhibitors from Finger Millet (Eleusine coracana).

Hypertension remains a significant global health concern, contributing significantly to cardiovascular diseases and mortality rates. The inhibition of angiotensin-converting enzyme (ACE) plays a crucial role in alleviating high blood pressure. We investigated the potential of finger millets (Eleusine coracana) as a natural remedy for hypertension by isolating and characterizing its ACE-inhibitory compound. First, we evaluated the ACE-inhibitory activity of the finger millet ethanol extract and subsequently proceeded with solvent fractionation. Among the solvent fractions, the ethyl acetate fraction exhibited the highest ACE inhibitory activity and was further fractionated. Using preparative high-performance liquid chromatography, the ethyl acetate fraction was separated into four subfractions, with fraction 2 (F2) exhibiting the highest ACE inhibitory activity. Subsequent 1H-nuclear magnetic resonance (NMR) and 13C-NMR analyses confirmed that the isolated compound from F2 was catechin. Furthermore, molecular docking studies indicated that catechin has the potential to act as an ACE inhibitor. These findings suggest that finger millets, particularly as a source of catechin, have the potential to be used as a natural antihypertensive.

Identification of Pheophytin a and Hydroxy Pheophytin a from Rang Chuet (Thunbergia laurifolia Linn.) as Potent NQO-1 Inducers in Liver Cells.

Thunbergia laurifolia Linn. (Rang Chuet, RC), a Thai medicinal plant, possesses various bioactive compounds with potential health benefits. This study aimed to identify detoxifying compounds within RC crude extract. RC leaves were extracted using the Soxhlet method with chloroform. Total carotenoids, chlorophylls, extract yield, total phenolic contents (TPCs), and total flavonoid contents (TFCs) were measured. The extract's composition was analyzed. Cytotoxicity and effects on the detoxification enzyme NQO-1 were assessed in liver cell lines (AML12 and HepG2) using MTT and NQO-1 assays, respectively. Bioactive fractions were identified using fractionation techniques and mass spectrometry (LC-MS). RC extract displayed significant levels of carotenoids (0.375 mg/g), chlorophylls (2.682 mg/g), and favorable yield (15.3%). TPC and TFC were 363.776 mg/g and 112.22 mg/g of extract, respectively. Analysis revealed phenolic acids (gallic acid, caffeic acid), flavonoid (apigenin), chlorophylls (chlorophylls a, b, pheophytin a and b), and lutein. Among the fractions, Fraction 3 (F3) exhibited the highest NQO-1 enzyme activity. F3 contained pheophytin a and hydroxy pheophytin a, confirmed by LC-MS (m/z 871.59+ [M + H]+ and 887.59+ [M + H]+). F3 significantly induced NQO-1 activity in both HepG2 (3.908-fold) and AML12 (1.99-fold) cells. This study identified F3 from RC extract as a promising fraction containing pheophytin a and hydroxy pheophytin a, responsible for inducing the detoxification enzyme NQO-1 in liver cells. These findings suggest RC's potential for promoting detoxification.

Evaluasi Kajian Oil Extraction Rate (OER) Optimum sebagai Standar Panen Kelapa Sawit

This research aims to carry out a basic evaluation of the harvest of palm kernel 3 (BD3), palm kernel 7 (BD7), fraction 1 (FR1), and fraction 2 (FR2) on the types of soil and peat and minerals that are tested and analyzed based on observations of the number of loose fruits from bunches (brondolan) on the plate. This research was conducted at PT Gandaera Hendana, Ukui 2 Village, Ukui District, Pelalawan Regency, Riau Province. The study used a factorial randomized block design (RBD) with two factors: harvest base and two types of soil. The parameters observed consisted of adding a number of brondolan after harvest, the multiple of brondol after harvest, oil to bunch analysis, and potential oil extraction rate (OER). The analysis showed that adding loose fruit after harvest in mineral soil increased significantly two times more than peat (15.2 loose fruit). The harvest standard for fraction 2 increased significantly due to the addition of loose bunches after harvest (23.9 bunches) and their interaction in mineral soil (38.6 bunches). The base of palm kernels 3 (BD ​​3) shows a very significant increase in the yield of palm kernels after harvest (2.4 times) as well as its interaction in mineral soil (2.7 times). The palm kernels harvest standard 3 showed the highest increase in OER potential (23.25%) and OER potential in mineral soil (24.46%).

Co-pigmentation of Pinotage anthocyanins with Cyclopia (honeybush) polyphenols in model wine solutions

Co-pigmentation of an anthocyanin-enriched fraction (AEF) of Pinotage wine with honeybush polyphenols (Cyclopia subternata extract and fractions enriched in specific polyphenols) was studied in model wine solutions. The hyperchromic effects and bathochromic shifts depended on the co-pigment sources and co-pigment/anthocyanin ratio. Fraction 2 (F2), enriched in xanthones (mangiferin and isomangiferin), produced the most pronounced co-pigmentation effect at a co-pigment/anthocyanin ratio of 15:1. Greater co-pigmentation effects were observed in model solutions at pH 3.0 (pH 3–4 evaluated) and in the absence of EtOH (0–16 % EtOH evaluated). During storage of solutions (AEF and/or F2) at 15 °C and 25 °C for 12 weeks, myrtillin, petunin, 3′,5′-di-β-D-glucopyranosyl-3-hydroxyphloretin and mangiferin concentrations decreased following first-order kinetics, while peonin and oenin degradation followed fractional-conversion kinetics. Polymeric anthocyanin formation followed zero-order kinetics. Reaction rates increased with an increase in storage temperature. The addition of F2 to the AEF solution produced a darker, more vivid colour and improved the colour stability, however, the yellow-orange colour of F2 and intensification of its colour during storage contributed to increasing red-orange hues. Further research is necessary to elucidate the effect of honeybush polyphenols on the colour and composition of a complete wine as it is more complex than model solutions.

Antimicrobial Activities of Some Constituents Isolated from a Kenyan Medicinal Plant, Capparis fascicularis DC

Background of the Study. Capparis fascicularis DC. is an indigenous medicinal plant belonging to the family Capparaceae found in Nyandarua County, Nairobi, Kenya, and many parts of Africa. It is a shrub whose roots are used traditionally to treat colds. Aim. The aim of the present study is to carry out antimicrobial activities of solvent extracts of different parts of Capparis fascicularis, characterize the phytochemical constituents of the most active extract, and identify the most active compounds. Place and Duration of the Study. All the experiments were carried out in the departments of Chemistry and Microbiology, Kenyatta University, Nairobi, Kenya. Methodology. Petroleum ether, ethyl acetate, methanol, and water extracts from the leaves, stem bark, and root bark of C. fascicularis were screened against selected strains of both Gram-positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative (Salmonella typhimurium and Escherichia coli) bacteria using disc diffusion and tube dilution methods. Fractionation of the most active crude extract was carried out by column chromatography, and the fractions together with the most active crude extract were screened against the selected bacterial strains. The most active fraction was further fractionated, and the subfractions were screened against the bacterial strains to test for possible synergistic effects between the subfractions and their constituents. The most active fraction was then analysed by GC-MS and LC-ESI-MS methods to identify the major constituents. Results. The ethyl acetate extract of C. fascicularis root bark (CFR2) showed a significant in vitro antibacterial activity. From the seven fractions of CFR2 obtained, fraction 2 (F2) had the lowest MIC value and was thus most active. Moreover, F2 was found to be more active compared to the four subfractions obtained from it. This suggested that constituents of F2 worked in synergy. Fraction 2 contained phenols, terpenes, and flavonoids. Two compounds were identified by GC-MS as 2,4-di-tert-butylphenol and (E,E)-2,4-decadienal, while three compounds were identified by LC-ESI-MS as tanshinone II A, cryptotanshinone, and danshensu. Conclusion. The study revealed that CFR2 is the most active extract on bacteria, suggesting that most antimicrobial compounds are concentrated in the roots of C. fascicularis. A follow-up study is directed towards chromatographic separations to obtain the other chemical constituents and screen them against various strains of bacteria.

Cytotoxic properties of unfractionated and fractionated bromelain alone or in combination with chemotherapeutic agents in colorectal cancer cells.

Colorectal cancer (CRC) is one of the most lethal cancers worldwide. Long-term survival is not achieved in metastatic CRC despite the current multidisciplinary therapies. Bromelain, a compound extracted from the pineapple plant, has multiple functions and anticancer properties. Previously, bromelain has been chromatographically separated into four fractions. Fraction 3 (F3) exhibits the highest proteolytic activity. The anticancer effects of F3 bromelain in CRC cells is unknown. In vitro cytotoxicity was verified through a sulforhodamine B assay. Apoptosis in CRC cells induced by unfractionated or F3 bromelain was examined using Annexin V-FITC/PI staining and Western blot analysis. ROS status, autophagy and lysosome formation were determined by specific detection kit. The cytotoxicity of F3 bromelain in CRC cells was found to be comparable to that of unfractionated bromelain. F3 bromelain induces caspase-dependent apoptosis in CRC cells. Treatment with unfractionated or F3 bromelain increased superoxide and oxidative stress levels and autophagy and lysosome formation. ATG5/12 and beclin-1 were upregulated, and the conversion of LC3B-I to LC3B-II was increased significantly by treatment with F3 bromelain. Treated CQ, autophagy inhibitor, with unfractionated or F3 bromelain enhances the cytotoxic effects. Finally, the combination of unfractionated and F3 bromelain with a routine chemotherapeutic agent (5-fluourouracil, irinotecan, or oxaliplatin) resulted in synergistically higher cytotoxic potency in CRC cells. Unfractionated and F3 bromelain inhibits CRC cell proliferation in vitro, and the cytotoxic effects of unfractionated bromelain are equivalent to F3 bromelain. F3 bromelain may be a potential and potent drug for clinical use due to its anticancer efficacy and high synergistic cytotoxicity when combined with a routine chemotherapeutic agent for CRC.

The impact of psyllium gelation behaviour on in vitro colonic fermentation properties

Psyllium is a viscous, gel forming fibre with properties that have led it to be used for alleviating gastrointestinal discomfort. We have used previously identified fractions of psyllium with differing flow properties. Fraction 1 (F1) forms a non-gelling solution containing rhamnose. galactose, and arabinose. Fraction 2 (F2) forms a fluid-like gel containing mainly xylose and arabinose, Fraction 3 (F3) has almost identical monosaccharide and linkage composition to F2, but forms an insoluble, self-supporting gel. We performed in vitro batch fermentation experiments seeded with human stool. Metabolomics were performed using 1H NMR, and FISH with calcofluor white and direct red 23 were used to visualise the gels after in vitro fermentation of the fractions. The total amount of gas and short chain fatty acid produced was significantly higher for F1, compared to F2 and F3. F3 gas production was significantly lower than F2, but metabolite production between F2 and F3 did not differ. All fractions preferentially lead to the production of propionate instead of butyrate and were produced in the ratio of 58:35:7, 54:38:8, and 61:33:6 (acetate: propionate: butyrate) for F1, F2, and F3 respectively. Microscopy showed differences in how the fractions broke down and demonstrated the localisation of bacteria on the outer edge of each fraction. These results suggest that for these psyllium fractions the structure is a key factor that determines fermentability. Flow properties may play a role in gas production, suggesting directions for future investigation. Isolated fractions may have clinical benefit above that of unrefined psyllium powder aiding in the treatment of gastrointestinal discomfort.

Ethnomedicinal Plant Stephania hernandifolia and its Active Fraction 2- Chloroethyl Linoleate Inhibits HSV-2 Infection by Blocking Viral Immediate Early and Early Transcription

Aim: The aim of this study is validation of antiviral activity of Stephania hernandifolia against HSV-2. Background: Ethnomedicinal plant Stephania hernandifolia, traditionally used for the management of skin, digestive and nerve ailments demonstrated significant anti-HSV-1 activity; similar to Stephania cepharantha having neuroinflammatory and anti-HSV activities. Objectives: Thus, the present study aimed to validate the potential of the most active fraction-2 (F-2) of S. hernandifolia against HSV-2 in vitro, along with the underlying mode or mechanism of action. Methods: The standardized F-2 was characterized by GC-MS, 1H-NMR, Mass and FTIR spectroscopy. Cytotoxicity (CC50) and antiviral activity (EC50) were evaluated by MTT and Plaque reduction assay. To determine the mode of action, we have used time-of-addition, virus inactivation, and entry (attachment and penetration) assays, followed by semiquantitative PCR. Furthermore, the protein expression levels of immediate early (IE) and early (E) gene products of drug-treated virions were measured by Western blot. Results: The results showed that HSV-2G and ICMR/VU-2012/20, the clinical isolate of HSV-2, were inhibited by F-2 at EC50 of 20.0 and 20.43 μg/ml respectively, with Selectivity Index (SI) of 12. Timeof- addition assay showed that F-2 significantly inhibited HSV-2 infection in Vero cells at 4-8 h posttreatment. The infectivity of the virion was lost within 1h of exposure to F-2 (EC50 and EC99). Furthermore, semi-Q-PCR and Western blot studies demonstrated significant downregulation of IE and E gene products. The characterization revealed that 2-chloroethyl linoleate is the lead compound in the F-2 fraction. Conclusion: Thus, our results showed that the bioactive fraction F-2 inhibits both IE and E transcription of HSV-2.

Purification and identification of novel antioxidant peptides from silkworm pupae (Bombyx mori) protein hydrolysate and molecular docking study

Crude proteins from the pupae of two indigenous strains of silkworm (Bombyx mori), Nangnoi and Nangsew were hydrolyzed by alcalase, papain and trypsin. Protein hydrolysate of Nangnoi strain prepared by alcalase showed the highest antioxidant activities. In addition, the DNA damage protection ability of this protein hydrolysate was evaluated. Subsequently, this protein hydrolysate was purified by passing through a 3 kDa MW cut-off membrane followed by reverse phase high performance liquid chromatography (RP-HPLC). Amongst eight fractions obtained after RP-HPLC purification, fraction S3 showed interesting antioxidant activities with the IC50 values toward ABTS and DPPH radicals of 4.78 ± 0.08 and 7.83 ± 0.02 μg/mL, respectively. Potentially antioxidant peptides, AAEYPA and AKPGVY were identified in the S3 fraction after analysis by LC-MS/MS. ABTS and DPPH radicals scavenging activities of both synthesized peptides were investigated. These results suggest that both hexapeptides are potential antioxidants that can be used in the food and cosmetic industries.

Extraction optimization and screening of antioxidant peptides from grass carp meat and synergistic-antagonistic effect.

Grass carp (Ctenopharyngodon idellus) is one of the three most cultivated freshwater fish around the world, but it is mainly consumed afresh, so only a small part of them are processed into salted fish or snack food. This research was performed to prepare and screen antioxidant peptides from grass carp muscle to promote its high‐value utilization. The parameters of double‐enzyme two‐step hydrolysis were optimized, the peptides with the highest ABTS.+ scavenging ability were enriched and identified by Sephadex G‐25 and LC‐Q‐Orbitrap‐MS/MS. The synergistic–antagonistic effect among identified peptides was also investigated. The optimized conditions were hydrolyzed with protamex (10,000 U/g) at pH 8.0, 50°C for 3 h, followed by hydrolysis with alcalase (6,000 U/g) at pH 9.0, 50 °C for 2 h, and the protein–liquid ratio was 4%. The hydrolysates were further fractionated to obtain five fractions, in which fraction 3 (F3) exhibited the strongest ABTS.+ and O2·‐ scavenging ability with the IC50 values of 0.11 and 0.47 mg/ml, respectively. Twelve novel antioxidant peptides were identified, in which VAGW possessed the highest activity (139.77 μmol GSH/g). Significantly synergistic effects were observed on the two and three peptides’ combination among VAGW, APPAMW, LFGY, FYYGK, and LLLYK, while the C‐terminal tryptophan (Trp) played an important role in the synergism. This study found that grass carp muscle hydrolysates can be potential natural antioxidants in functional products. The synergistic effects among peptides may provide a perspective for the combined application of peptides.

Anti-breast cancer synthetic peptides derived from the Anabastestudineus skin mucus fractions

Previous study has shown the antimicrobial activities of mucus protein extracted from Anabastestudineus. In this study, we are interested in characterizing the anticancer activity of the A.testudineus antimicrobial peptides (AMPs). The mucus was extracted, fractioned, and subjected to antibacterial activity testing to confirm the fish's AMPs production. The cytotoxic activity of each fraction was also identified. Fraction 2 (F2), which shows toxicity against MCF7 and MDA-MB-231 were sent for peptide sequencing to identify the bioactive peptide. The two peptides were then synthetically produced and subjected to cytotoxic assay to prove their efficacy against cancer cell lines. The IC50 for AtMP1 against MCF7 and MDA-MB-231 were 8.25 ± 0.14 μg/ml and 9.35 ± 0.25 μg/ml respectively, while for AtMP2 it is 5.89 ± 0.14 μg/ml and 6.97 ± 0.24 μg/ml respectively. AtMP1 and AtMP2 treatment for 48 h induced breast cancer cell cycle arrest and apoptosis by upregulating the p53, which lead to upregulate pro-apoptotic BAX gene and downregulate the anti-apoptotic BCL-2 gene, consequently, trigger the activation of the caspase-3. This interaction was supported by docking analysis (QuickDBD, HPEPDOCK, and ZDOCK) and immunoprecipitation. This study provided new prospects in the development of highly effective and selective cancer therapeutics based on antimicrobial peptides.

Antinociceptive effects of flower extracts and the active fraction from Styrax japonicus

Ethnopharmacological relevanceFlowers from Styrax japonicus sieb. et Zucc. have been used as a Chinese folk medicine to alleviate pain such as toothache and sore throat. Aim of the studyTo testify the analgesic effect of flowers from Styrax japonicus, analyze components of the active fraction, and investigate the mechanism of analgesia. Materials and methodsFlower extracts were obtained by ethanol, petroleum ether and hydrodistillation extraction. Different fractions of ethanol extracts (EE) were isolated by silica gel column chromatography and preparative liquid chromatography. Analgesic effects of EE, petroleum ether extracts (PEE), hydrodistillation extracts (HDE), and fractions of EE were evaluated using hot plate, acetic acid-induced writhing and formalin tests on mice. Components of the active fraction 1 (F1) were determined by the ultrahigh-performance liquid chromatography Q extractive mass spectrometry (UHPLC-QE-MS). Anti-inflammatory and sedative effects involving analgesic mechanisms were evaluated by carrageenan induced hind paw oedema and pentobarbital sodium sleep tests, respectively. In addition, antagonists including naloxone hydrochloride (NXH), flumazenil (FM), SCH23390 (SCH) and WAY100635 (WAY) were used to investigate the possible mechanism of analgesia. Contents of neurotransmitters and relevant metabolites in different brain regions of mice were also quantified by the ultraperformance liquid chromatography with a fluorescence detector (UPLC-FLD). ResultsEE rather than PEE and HDE at medium and high doses (150 mg/kg and 300 mg/kg) significantly prolonged the latency time of the response of mice to the thermal stimulation in the hot plate test. Moreover, EE significantly decreased number of writhes in the acetic acid-induced writhing test, and reduced licking time in both two phases of the formalin test in a dose-dependent manner. The F1 (50 mg/kg) showed effective antinociceptive responses in all mice models. However, fraction 2 (F2) and fraction 3 (F3) at 50 mg/kg performed no analgesic action. Kaempferol-3-O-rutinoside, isorhamnetin-3-O-rutinoside, pinoresinol-4-O-glucoside, forsythin and arctiin were identified from components of the F1. Furthermore, F1 (50 mg/kg) did not significantly affect hind paw oedema of mice induced by carrageenan but significantly shortened sleep latency and increased sleep duration in the pentobarbital sodium sleep test. In addition, the antinociceptive response of F1 was not affected by NXH in two mice models, but significantly blocked by FM and WAY in the hot plate test. In the formalin test, FM avoided the effect of F1 only in the first phase, while the analgesic activity of F1 was totally suppressed by WAY in both two phases. Otherwise, contents of 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) increased significantly in hippocampus and striatum of mice in the F1 group. ConclusionEE from flowers of Styrax japonicus, and F1, the active part isolated from EE, showed significant antinociceptive activities. The analgesic effect of F1 appeared to be related to the sedative effect, partially mediated by the GABAergic system, and highly involved in the serotonergic system. This was the first study confirming the analgesic effect of Styrax japonicus flower, which provided a candidate for the development of non-opioid analgesics.

Fractionating of Lemongrass (Cymbopogon citratus) Essential Oil by Vacuum Fractional Distillation

Lemongrass essential oil has many compounds appropriate for application in foods, cosmetics, and pharmaceutical products. Of these, citral is a high-value compound of interest to industry. This work aims to evaluate the use of vacuum fractional distillation to separate lemongrass essential oil compounds, producing essential oil fractions containing high citral content. The effect of process parameters, namely vacuum pressure, type column, and energy input, on the fractionation time, content, and recovery of citral in the fractions, was investigated. The fractionation of lemongrass oils successfully provided five fractions, i.e., fraction 1 (F1), fraction 2 (F2), fraction 3 (F3), fraction 4 (F4), and fraction 5 (F5). GC-MS (Gas Chromatography-Mass Spectrometry) spectra showed that the main compound contained in F1 and F2 fractions was β-myrcene (>70%). Meanwhile, F4 and F5 were the two main fractions for citral recovery. The optimal conditions of the fractional distillation system included a column height of 400 mm, power input of 165 W, and pressure of 15 mmHg. These conditions correspond to the highest total citral content of 95%, with a recovery of 80% at the F4 and F5 fractions. Therefore, fractional vacuum distillation may be an effective method to upgrade lemongrass essential oil.

Antidiabetic Property of a Fraction of the Methanol Extract of the Seeds of Abrusprecatorius in Alloxan-induced Diabetic Rats

Introduction: Diabetes mellitus (DM) is a group of metabolic diseases characterized by hyperglycemia resulting from defects of insulin secretion and/or increased cellular resistance to insulin. Abrusprecatoriushas been used for the treatment of various ailments including diabetes mellitus. This study is aimed at evaluating the possible anti-diabetic effect of the methanol fraction 2 (F2) of the seeds of Abrusprecatoriusin alloxan-induced diabetic Wistar albino rats.
 Methods: The methanol extract of the seeds of A. precatorius Linn Fabaceae was fractionated by Sephadex G15. Diabetes was induced by a single intraperitoneal administration of 150 mg/kg bodyweight of alloxan. Acute toxicity (LD50) study was done using Lorke’s method. The antidiabetic activities of Fraction 2 (F2) and the biochemical parameters were investigated using standard diagnostic methods.
 Results: The value of the LD50 is 529.2 mg/kg bw. Oral doses (5, 10 and 20 mg/kg bw.) of the extract caused a dose-dependent significant (p<0.05) reductions in blood glucose concentrations compared with diabetic untreated controls. There were significant decreases (p<0.05) in the total cholesterol and triacylglycerol levels of rats in the treatment groups compared with the diabetic untreated group. The groups that were treated with glibenclamide, 5 mg/kg of the fraction, 20mg/kg of the fraction and the group that was pre-treated with 10mg/kg of the fraction before the induction of diabetes showed a significant decrease (p<0.05) in the concentration of low-density lipoprotein and significant increase (p<0.05) in high-density lipoprotein concentrations compared with the diabetic untreated group. The protein level increased non-significantly (p>0.05) in all the test groups compared with the diabetic untreated control group except the pre-treated group which increased significantly (p<0.05).
 Conclusion: The results obtained from this study revealed that the methanol fraction of A. precatorius has antidiabetic property which can be as a result of the important phytochemicals found in the F2 fraction. Fraction 2 (F2) however showed improvement in lipid profile.

Effects of a Fraction of the Methanol Extract of the Seeds of Abrus Precatorius on Malondialdehyde and Antioxidant Levels of Alloxan-Induced Diabetic Rats

Introduction: The aim of this study is to evaluate the effect of fraction 2 (F2) of methanol extract of the seeds of Abrus precatorius on malondialdehyde and antioxidant level of alloxan-induced diabetic Wistar albino rats.
 Materials and Methods: The methanol extract of the seeds of A. precatorius Linn Fabaceae was fractionated by Sephadex G15. Diabetes was induced by a single intraperitoneal administration of alloxan at a dose of 150 mg/kg bodyweight. The phytochemical analysis and the biochemical parameters were investigated using standard diagnostic methods.
 Results: Both qualitative and quantitative analysis of F2 revealed the presence of high amounts of alkaloids (2000 ± 80 mg/100 g), flavonoids (158 ± 17.6 mg/100 g) and tannins (258 ± 45 mg/100 g) but low concentration of saponins (18.3 ± 2.43 mg/100 g). The malondialdehyde (MDA) level of all the test groups and the group treated with the standard drug decreased significantly (p<0.05) compared with that of untreated diabetic group. However, the catalase activity in all the test groups significantly increased (p<0.05) while the activity of superoxide dismutase increased significantly (p<0.05) in groups administered 20mg/kg of the fraction and the group pre-treated with 10mg/kg of the fraction compared with the diabetic untreated group. Similarly, the result revealed a significant increase (p<0.05) in non-enzymatic antioxidants such as reduced glutathione and vitamin E of the rats treated with graded doses of F2 while vitamin C showed significant (p<0.05) increase in normal control groups and pre-treated groups compared with the diabetic untreated. Studies on membrane stabilization using hypotonicity-induced red blood cell haemolysis revealed that the F2 of the methanol extract of the seeds of A. precatorius Linn inhibited haemolysis in a dose-dependent manner.
 Conclusion: The results obtained from this study revealed that the F2 fraction of the methanol extract of A. precatorius contain important phytochemicals found in the Fraction 2 (F2) however showed improvement in antioxidant defence as well as has a stabilizing effect on the membrane.

Bioactive Fraction of Mentha piperita Leaves Extract Stimulates Lipoprotein Lipase at Cellular and Molecular Levels via Augmentation of Antioxidant Defense System in Hyperlipidemic Rats

ABSTRACT The antihyperlipidemic and antioxidant effects of Mentha piperita leaves extracts in Poloxamer 407-induced hyperlipidemic rats were investigated. Antihyperlipidemic activities of hexane, ethyl acetate, and methanol extracts (100, 200, or 300 mg kg−1 b.w.) were evaluated by lipid profile analysis. Twelve (12) fractions (F1 to F12) were collected from the most antihyperlipidemic extract (ethyl acetate at 100 mg kg−1) and, assayed in vivo for antihyperlipidemic activity. The effects of the most antihyperlipidemic fraction (F2) on redox sensitive biomarkers (SOD, CAT, GSH, MDA) and lipoprotein lipase (LPL) activity at cellular and molecular levels were evaluated. Fraction-2 (F2) was further subjected to GC-MS and LC-MS analysis for identification of nonpolar and polar components. The F2 treatment decreased the level of MDA with concomitant enhancement of GSH, LPL activity in the liver and serum and up regulated the expression of SOD, CAT, and LPL genes in liver and blood relative to the untreated rats. GC-MS and LC-MS revealed oleic acid, vaccenic acid, 3, 4-dicaffeoylquinic acid, coumarin, and triacontane as the probable bioactive components with reported antihyperlipidemic and antioxidant effects.

Cytotoxic Activity Against L1210 Leukemia Cells from the Ethyl Acetate Fraction of Kenikir Leaves (Cosmos. Caudatus) Preserved by Gamma Irradiation

<p><em>K</em><em>enikir</em> leaves (<em>Cosmos caudatus</em>) Has been used as traditional medicine, especially as an anti-cancer, this plant has been in Indonesia both in herbs and capsules. Microbial contamination of herbal medicinal raw materials occurs when the storage process is done. One of the preservation techniques used in the industry is using gamma irradiation techniques to reduce microbial and fungal contamination. The purpose of this research was to study the effect of gamma irradiation for the preservation of <em>kenikir</em> leaves (<em>C</em><em>.</em><em> caudatus</em>) as an anti-cancer based on cytotoxic activity using L1210 leukaemia cells. The simplicia was gamma-irradiated by Co-60 source with variation doses of 0 (control); 5; 7.5; 10; and 15 kGy. Then the irradiated and control samples were macerated successively using <em>n</em>-hexane, ethyl acetate, and ethanol. The active extract (ethyl acetate) was further fractionated using column chromatography, obtained seven fractions (F1 - F7). The seven fractions' cytotoxic activity against L1210 leukaemia cells showed that fraction 3 (F3) was the most active fraction with an IC<sub>50</sub> of 1.26 µg/mL. Each dose's F3 cytotoxic activity showed that the IC50 7.5 kGy irradiation sample did not change significantly with control (0 kGy) based on ANOVA analysis using SPSS 24 with a 95% confidence level. In comparison, irradiation samples of 10 and 15 kGy showed a change in the IC<sub>50</sub> value is significant with the control (0 kGy). These results indicate that gamma irradiation can be used as an alternative for preserving <em>C. caudatus</em> with a maximum dose of 7.5 kGy, so that its anti-cancer properties do not change with those without irradiation.</p>

Characterisation and Hypolipidaemic Activity of Phenylquinoline, and Narceine Isolated from Ficus polita Leaf

Aim: To evaluate hypolipidaemic potentials of column chromatography fractions (F1 to F6) of the chloroform leaf extract of Ficus polita and to detect the bioactive compounds present in the most active fraction using spectroscopic techniques.
 Study Design: Fourty-five (45) wistar rats were grouped into nine groups of five rats each: normal control, hyperlipidaemic control, hyperlipidaemic administered with standard drug control/ atorvastatin (10 mg/kg body weight), and hyperlipidaemic administered groups administered with 50 mg/kg body weight of column chromatography fractions (F1, F2, F3, F4, F5, and F6) for a period of two weeks.
 Place and Duration of Study: Department of Biochemistry Laboratory, Faculty of Basic Medical Sciences, Bayero University Kano, Nigeria, from May 2018 to April 2019.
 Methodology: Chloroform leaf extract of F. polita was fractionated by using column chromatography, and the resulting fractions were pooled, based on their retention factor (Rf), into six (6) fractions by using analytical thin layer chromatography. The resulting six (6) fractions were screened for hypolipidaemic activity. Serum total cholesterol (TC), triglycerides (TG), high density lipoprotein-cholesterol (HDL), and low density lipoprotein cholesterol (LDL) were determined. The rats treated with the best fraction in terms hypolipidaemic activity (fraction 3) were screened for serum HMG Co A reductase, lactate dehydrogenase (LDH), creatine kinase (CK) activities, as well as troponin I level. Oxidative stress markers such as malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) were determined in the heart tissue homogenate of the rats treated with fraction 3.
 Results: Fraction 3 treated hyperlipidaemic group showed significant (p<0.05) decrease in the levels of serum TC, TG, and LDL, but significant (p<0.05) increase in the level of serum HDL. Fraction 3 (F3) treated hyperlipidaemic groups showed significant (p<0.05) decrease in the activity of serum LDH and the level of troponin I, but significant (p<0.05) increase in the activity of serum HMG Co A reductase. After high fat diet administration, the level of heart tissue antioxidant markers such as GSH, SOD and catalase were decreased whereas the level of heart tissue MDA was elevated. The level of these antioxidant markers were brought to normalcy by fraction 3 (F3). Histological studies of the heart corroborated the biochemical findings, and treatment with fraction 3 (F3) was found to be effective in restoring dietary-induced myocardiac toxicity in rats. FTIR and GCMS analyses were carried out for the detection of bioactive compound(s) in fraction 3 (F3), and the result revealed the presence of "8-methoxy-4-phenylquinoline, and narceine.
 Conclusion: The study concludes that; the hypolipidaemic property of the leaf of F. polita is mediated by the bioactive compounds "8-methoxy-4-phenylquinoline, and narceine." via their antioxidant properties.