Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 6910

Search results for: energy metabolites

6910 Gut Metabolite Profiling of the Ethnic Groups from Assam, India

Authors: Madhusmita Dehingia, Supriyo Sen, Bhuwan Bhaskar, Tulsi Joishy, Mojibur R. Khan

Abstract:

Human gut microbes and their metabolites are important for maintaining homeostasis in the gut and are responsible for many metabolic and immune mediated diseases. In the present study, we determined the profiles of the gut metabolites of five different ethnic groups (Bodo, Tai-Phake, Karbi, Tea tribe and Tai-Aiton) of Assam. Fecal metabolite profiling of the 39 individuals belonging to the ethnic groups was carried out using Gas chromatography – Mass spectrometry (GC-MS), and comparison was performed among the tribes for common and unique metabolites produced within their gut. Partial Least Squares Discriminant Analysis (PLS-DA) of the metabolites suggested that the individuals grouped according to their ethnicity. Among the 66 abundant metabolites, 12 metabolites were found to be common among the five ethnic groups. Additionally, ethnicity wise some unique metabolites were also detected. For example, the tea tribe of Assam contained the tea components, Aniline and Benzoate more in their gut in comparison to others. Metabolites of microbial origin were also correlated with the already published metagenomic data of the same ethnic group and functional analysis were carried out based on human metabolome database.

Keywords: ethnicity, gut microbiota, GC-MS, metabolites

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6909 Secondary Metabolites from Turkish Marine-Derived Fungi Hypocrea nigricans

Authors: H. Heydari, B. Konuklugil, P. Proksch

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Marine-derived fungi can produce interesting bioactive secondary metabolites that can be considered the potential for drug development. Turkey is a country of a peninsula surrounded by the Black Sea at the north, the Aegean Sea at the west, and the Mediterranean Sea at the south. Despite the approximately 8400 km of coastline, studies on marine secondary metabolites and their biological activity are limited. In our ongoing search for new natural products with different bioactivities produced by the marine-derived fungi, we have investigated secondary metabolites of Turkish collection of the marine sea slug (Peltodoris atromaculata) associated fungi Hypocrea nigricans collected from Seferihisar in the Egean sea. According to the author’s best knowledge, no study was found on this fungal species in terms of secondary metabolites. Isolated from ethyl acetate extract of the culture of Hypocrea nigricans were (isodihydroauroglaucin,tetrahydroauroglaucin and dihydroauroglaucin. The structures of the compounds were established based on an NMR and MS analysis. Structural elucidation of another isolated secondary metabolite/s continues.

Keywords: Hypocrea nigricans, isolation, marine fungi, secondary metabolites

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6908 Heroin and Opiates Metabolites Tracing by Gas-Chromatography Isotope Ratio Mass Spectrometry

Authors: Yao-Te Yen, Chao-Hsin Cheng, Meng-Shun Huang, Shan-Zong Cyue

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'Poppy-seed defense' has been a serious problem all over the world, that is because the opiates metabolites in urine are difficult to distinguish where they come from precisely. In this research, a powerful analytic method has been developed to trace the opiates metabolites in urine by Gas-Chromatography Isotope Ratio Mass Spectrometry (GC-IRMS). In order to eliminate the interference of synthesis to heroin or metabolism through human body, opiates metabolites in urine and sized heroin were hydrolyzed to morphine. Morphine is the key compound for tracing between opiates metabolites and seized heroin in this research. By matching δ13C and δ15N values through morphine, it is successful to distinguish the opiates metabolites coming from heroin or medicine. We tested seven heroin abuser’s metabolites and seized heroin in crime sites, the result showed that opiates metabolites coming from seized heroin, the variation of δ13C and δ15N for morphine are within 0.2 and 2.5‰, respectively. The variation of δ13C and δ15N for morphine are reasonable with the result of matrix match experiments. Above all, the uncertainty of 'Poppy-seed defense' can be solved easily by this analytic method, it provides the direct evidence for judge to make accurate conviction without hesitation.

Keywords: poppy-seed defense, heroin, opiates metabolites, isotope ratio mass spectrometry

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6907 The Order Russulales of Basidiomycota: Systematics, Ecology and Chemotaxonomy

Authors: Marco Clericuzio, Alfredo Vizzini

Abstract:

The secondary metabolites of Russulales (one of the main orders of phylum Basidiomycota), have been studied. They are mainly terpenoids, with sesquiterpenes being the most common ones, but also triterpenoids and prenylated phenols have been isolated. We found that classes of specific compounds seem to be often allied to systematic groupings, so that they may have chemotaxonomic significance. Moreover, the ecological implications of such metabolites, as well as their biological activities, are discussed. Lately, we have focused our attention on the anti-arthropod activity of Russula metabolites, in particular on the toxicity against mites and other crop pests.

Keywords: chemotaxonomy, fungi, insecticidal activity, russulales, terpenoids

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6906 Energy Metabolites Show Cross-Protective Plastic Responses for Stress Resistance in a Circumtropical Drosophila Species

Authors: Ankita Pathak, Ashok Munjal, Ravi Parkash

Abstract:

Plastic responses to multiple environmental stressors in wet or dry seasonal populations of tropical Drosophila species have received less attention. We tested plastic effects of heat hardening, acclimation to drought or starvation; and changes in trehalose, proline and body lipids in D. ananassae flies reared under wet or dry season specific conditions. Wet season flies revealed significant increase in heat knockdown, starvation resistance and body lipids after heat hardening. However, accumulation of proline was observed only after desiccation acclimation of dry season flies while wet season flies elicited no proline but trehalose only. Therefore, drought-induced proline can be a marker metabolite for dry season flies. Further, partial utilization of proline and trehalose under heat hardening reflects their possible thermoprotective effects. Heat hardening elicited cross-protection to starvation stress. Stressor-specific accumulation or utilization, as well as rates of metabolic change for each energy metabolite, were significantly higher in wet season flies than dry season flies. Energy metabolite changes due to inter-related stressors (heat vs. desiccation or starvation) resulted in possible maintenance of energetic homeostasis in wet or dry season flies. Thus, low or high humidity induced plastic changes in energy metabolites can provide cross-protection to seasonally varying climatic stressors.

Keywords: wet-dry seasons, plastic changes, stress related traits, energy metabolites, cross protection

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6905 The Impact of Ramadan Fasting on Blood Pressure: Observational Study and a Meta-Analysis

Authors: Rami Al Jafar, Paul Elliott, Konstantinos K. Tsilidis, Abbas Dehghan

Abstract:

Although Ramadan fasting is a ritual that is practiced every year by millions of Muslims, studies on Ramadan fasting are still scarce. To the best of our knowledge, none of the previous studies comprehensively explored the effect on the metabolic profile. In London Ramadan Fasting Study, blood samples were collected from 81 participants before and 10-14 days after Ramadan. Blood samples were analysed using nuclear magnetic resonance (NMR) spectroscopy which covers 249 metabolites. Mixed-effects models were used to analyse collected data and assess the effect of Ramadan fasting on the metabolic profile. Our observational study involved 85 individuals with a mean age of 45.2 years, and 53.1% of them were males. After Ramadan, forty metabolites were affected significantly by Ramadan fasting. Most of these metabolites were metabolites ratios (24), and the rest were three Glycolysis, three ketone bodies, nine Lipoprotein subclasses and one inflammation marker. This study suggests that Ramadan fasting is significantly associated with changes in the metabolic profile. However, the changes are assumed to be temporary, and the long-term effect of these changes is unknown.

Keywords: metabolic profile, Ramadan fasting, metabolites, intermittent fasting

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6904 Identification and Characterization of in Vivo, in Vitro and Reactive Metabolites of Zorifertinib Using Liquid Chromatography Lon Trap Mass Spectrometry

Authors: Adnan A. Kadi, Nasser S. Al-Shakliah, Haitham Al-Rabiah

Abstract:

Zorifertinib is a novel, potent, oral, a small molecule used to treat non-small cell lung cancer (NSCLC). zorifertinib is an Epidermal Growth Factor Receptor (EGFR) inhibitor and has good blood–brain barrier permeability for (NSCLC) patients with EGFR mutations. zorifertinibis currently at phase II/III clinical trials. The current research reports the characterization and identification of in vitro, in vivo and reactive intermediates of zorifertinib. Prediction of susceptible sites of metabolism and reactivity pathways (cyanide and GSH) of zorifertinib were performed by the Xenosite web predictor tool. In-vitro metabolites of zorifertinib were performed by incubation with rat liver microsomes (RLMs) and isolated perfused rat liver hepatocytes. Extraction of zorifertinib and it's in vitro metabolites from the incubation mixtures were done by protein precipitation. In vivo metabolism was done by giving a single oral dose of zorifertinib(10 mg/Kg) to Sprague Dawely rats in metabolic cages by using oral gavage. Urine was gathered and filtered at specific time intervals (0, 6, 12, 18, 24, 48, 72,96and 120 hr) from zorifertinib dosing. A similar volume of ACN was added to each collected urine sample. Both layers (organic and aqueous) were injected into liquid chromatography ion trap mass spectrometry(LC-IT-MS) to detect vivozorifertinib metabolites. N-methyl piperizine ring and quinazoline group of zorifertinib undergoe metabolism forming iminium and electro deficient conjugated system respectively, which are very reactive toward nucleophilic macromolecules. Incubation of zorifertinib with RLMs in the presence of 1.0 mM KCN and 1.0 Mm glutathione were made to check reactive metabolites as it is often responsible for toxicities associated with this drug. For in vitro metabolites there were nine in vitro phase I metabolites, four in vitro phase II metabolites, eleven reactive metabolites(three cyano adducts, five GSH conjugates metabolites, and three methoxy metabolites of zorifertinib were detected by LC-IT-MS. For in vivo metabolites, there were eight in vivo phase I, tenin vivo phase II metabolitesofzorifertinib were detected by LC-IT-MS. In vitro and in vivo phase I metabolic pathways wereN- demthylation, O-demethylation, hydroxylation, reduction, defluorination, and dechlorination. In vivo phase II metabolic reaction was direct conjugation of zorifertinib with glucuronic acid and sulphate.

Keywords: in vivo metabolites, in vitro metabolites, cyano adducts, GSH conjugate

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6903 Parabens, Paraben Metabolites and Triclocarban in Sediment Samples from the Trondheim Fjord, Norway

Authors: Kristine Vike-Jonas, Susana V. Gonzalez, Olav L. Bakkerud, Karoline S. Gjelstad, Shazia N. Aslam, Øyvind Mikkelsen, Alexandros Asimakopoulos

Abstract:

P-hydrobenzoic acid esters (parabens), paraben metabolites, and triclocarban (TCC) are a group of synthetic antimicrobials classified as endocrine disrupting chemicals (EDCs) and emerging pollutants. The aim of this study was to investigate the levels of these compounds in sediment near the effluent of a wastewater treatment plant (WWTP) in the Trondheim Fjord, Norway. Paraben, paraben metabolites, and TCC are high volume production chemicals that are found in a range of consumer products, especially pharmaceuticals and personal care products (PCPs). In this study, six parabens (methyl paraben; MeP, ethyl paraben; EtP, propyl paraben; PrP, butyl paraben; BuP, benzyl paraben; BezP, heptyl paraben; HeP), four paraben metabolites (4-hydroxybenzoic acid; 4-HB, 3,4-dihydroxybenzoic acid; 3,4-DHB, methyl protocatechuic acid; OH-MeP, ethyl protocatechuic acid; OH-EtP) and TCC were determined by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) in 64 sediment samples from 10 different locations outside Trondheim, Norway. Of these 11 target analytes, four were detected in 40 % or more of the samples. The sum of six parabens (∑Parabens), four paraben metabolites (∑Metabolites) and TCC in sediment ranged from 4.88 to 11.56 (mean 6.81) ng/g, 52.16 to 368.28 (mean 93.89) ng/g and 0.53 to 3.65 (mean 1.50) ng/g dry sediment, respectively. Pearson correlation coefficients indicated that TCC was positively correlated with OH-MeP, but negatively correlated with 4-HB. To the best of the author’s knowledge, this is the first time parabens, paraben metabolites and TCC have been reported in the Trondheim Fjord.

Keywords: parabens, liquid chromatography, sediment, tandem mass spectrometry

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6902 Detection of Heroin and Its Metabolites in Urine Samples: A Chemiluminescence Approach

Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

Abstract:

A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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6901 Effect of Age and Physiological Status on Some Serum Energy Metabolites and Progesterone in Ouled Djellal Breed Ewes in Algeria

Authors: B. Safsaf, M. Tlidjane, B. Mamache, M. A. Dehimi, H. Boukrous, Aly A. Hassan

Abstract:

The aim of this study is to determine the effect of age and physiological status on progesterone and energy metabolism of Ouled Djellal (O.D) breed ewes. 40 healthy ewes were divided into two groups, primiparous and multiparous, with 20 ewes in each group. The body weights (BW) (kg) were 46.6 ± 4.20 and 59.2 ± 3.02, and consuming less 25 to 30% of their basal energetic requirements. The values of serum glucose, triglycerides and cholesterol were lower in pregnant than in non-pregnant ewes. The high to very high significant differences were found during the 15th week of pregnancy for glycaemia and triglyceridemia respectively. Concerning serum progesterone, a very highly significant difference (p < 0.001) was noted in the pregnant group, and the values were higher in MP than in PP. After lambing, the triglyceridemia values were slightly lower in primiparous than in multiparous pregnant ewes. In order to prevent imbalance during critical periods of reproduction, we can use the serum metabolic profile.

Keywords: age, energy metabolites, ouled djellal breed ewes, physiologic status, progesterone

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6900 Biosynthesis of Healthy Secondary Metabolites in Olive Fruit in Response to Different Agronomic Treatments

Authors: Anna Perrone, Federico Martinelli

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Olive fruit is well-known for the high content in secondary metabolites with high interest at nutritional, nutraceutical, antioxidant, and healthy levels. The content of secondary metabolites in olive at harvest may be affected by different water regimes, with significant effects on olive oil composition and quality and, consequently, on its healthy and nutritional features. In this work, a summary of several research studies dealing with the biosynthesis of healthy and nutraceutical metabolites of the secondary metabolism in olive fruit will be reported. The phytochemical findings have been correlated with the expression of key genes involved in polyphenol, terpenoid, and carotenoid biosynthesis and metabolism in response to different development stages and water regimes. Flavonoids were highest in immature fruits, while anthocyanins increased at ripening. In epicarp tissue, this was clearly associated with an up-regulation of the UFGT gene. Olive fruits cultivated under different water regimes were analyzed by metabolomics. This method identified several hundred metabolites in the ripe mesocarp. Among them, 46 were differentially accumulated in the comparison between rain-fed and irrigated conditions. Well-known healthy metabolites were more abundant at a higher level of water regimes. Increased content of polyphenols was observed in the rain-fed fruit; particularly, anthocyanin concentration was higher at ripening. Several secondary metabolites were differentially accumulated between different irrigation conditions. These results showed that these metabolic approaches could be efficiently used to determine the effects of agronomic treatments on olive fruit physiology and, consequently, on nutritional and healthy properties of the obtained extra-virgin olive oil.

Keywords: olea europea, anthocyanins, polyphenols, water regimes

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6899 Employing a Knime-based and Open-source Tools to Identify AMI and VER Metabolites from UPLC-MS Data

Authors: Nouf Alourfi

Abstract:

This study examines the metabolism of amitriptyline (AMI) and verapamil (VER) using a KNIME-based method. KNIME improved workflow is an open-source data-analytics platform that integrates a number of open-source metabolomics tools such as CFMID and MetFrag to provide standard data visualisations, predict candidate metabolites, assess them against experimental data, and produce reports on identified metabolites. The use of this workflow is demonstrated by employing three types of liver microsomes (human, rat, and Guinea pig) to study the in vitro metabolism of the two drugs (AMI and VER). This workflow is used to create and treat UPLC-MS (Orbitrap) data. The formulas and structures of these drugs' metabolites can be assigned automatically. The key metabolic routes for amitriptyline are hydroxylation, N-dealkylation, N-oxidation, and conjugation, while N-demethylation, O-demethylation and N-dealkylation, and conjugation are the primary metabolic routes for verapamil. The identified metabolites are compatible to the published, clarifying the solidity of the workflow technique and the usage of computational tools like KNIME in supporting the integration and interoperability of emerging novel software packages in the metabolomics area.

Keywords: KNIME, CFMID, MetFrag, Data Analysis, Metabolomics

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6898 Metabolomics Profile Recognition for Cancer Diagnostics

Authors: Valentina L. Kouznetsova, Jonathan W. Wang, Igor F. Tsigelny

Abstract:

Metabolomics has become a rising field of research for various diseases, particularly cancer. Increases or decreases in metabolite concentrations in the human body are indicative of various cancers. Further elucidation of metabolic pathways and their significance in cancer research may greatly spur medicinal discovery. We analyzed the metabolomics profiles of lung cancer. Thirty-three metabolites were selected as significant. These metabolites are involved in 37 metabolic pathways delivered by MetaboAnalyst software. The top pathways are glyoxylate and dicarboxylate pathway (its hubs are formic acid and glyoxylic acid) along with Citrate cycle pathway followed by Taurine and hypotaurine pathway (the hubs in the latter are taurine and sulfoacetaldehyde) and Glycine, serine, and threonine pathway (the hubs are glycine and L-serine). We studied interactions of the metabolites with the proteins involved in cancer-related signaling networks, and developed an approach to metabolomics biomarker use in cancer diagnostics. Our analysis showed that a significant part of lung-cancer-related metabolites interacts with main cancer-related signaling pathways present in this network: PI3K–mTOR–AKT pathway, RAS–RAF–ERK1/2 pathway, and NFKB pathway. These results can be employed for use of metabolomics profiles in elucidation of the related cancer proteins signaling networks.

Keywords: cancer, metabolites, metabolic pathway, signaling pathway

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6897 Phthalate Exposure among Roma Population in Slovakia

Authors: Miroslava Šidlovská, Ida Petrovičová, Tomáš Pilka, Branislav Kolena

Abstract:

Phthalates are ubiquitous environmental pollutants well-known because of their endocrine disrupting activity in human organism. The aim of our study was, by biological monitoring, investigate exposure to phthalates of Roma ethnicity group i.e. children and adults from 5 families (n=29, average age 11.8 ± 7.6 years) living in western Slovakia. Additionally, we analysed some associations between anthropometric measures, questionnaire data i.e. socio-economic status, eating and drinking habits, practise of personal care products and household conditions in comparison with concentrations of phthalate metabolites. We used for analysis of urine samples high performance liquid chromatography and tandem mass spectrometry (HPLC-MS/MS) to determine concentrations of phthalate metabolites monoethyl phthalate (MEP), mono-n-butyl phthalate (MnBP), mono-iso-butyl phthalate (MiBP), mono(2-ethyl-5-hydroxyhexyl) phthalate (5OH-MEHP), mono(2-ethyl-5-oxohexyl) phthalate (5oxo-MEHP) and mono(2-etylhexyl) phthalate (MEHP). Our results indicate that ethnicity, lower socioeconomic status and different housing conditions in Roma population can affect urinary concentration of phthalate metabolites.

Keywords: biomonitoring, ethnicity, human exposure, phthalate metabolites

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6896 Epicatechin Metabolites and Its Effect on ROS Production in Bovine Aortic Endothelial Cells

Authors: Nasiruddin Khan

Abstract:

The action of (-)-epicatechin, a cocoa (Theobroma cacao) flavanol that modulates redox/oxidative stress are contributed mainly to their antioxidant properties. The present study investigates the concentration and time dependent effect of (-)-epicatechin metabolites 3MeEc, 4MeEc, and 4SulEc on the production of ROS on BAEC using L-012, Lucigenin as chemiluminescence dye and XO/HX system. Our result demonstrates that 3MeEc shows significant (P <0.05) lowering effect of ROS production in BAEC with increasing concentration of metabolite while L-012 was used as chemiluminescence dye but not in the case of Lucigenin. In XO/HX system, using L-012 as chemiluminescence dye, 3MeEc and 4MeEc showed significant lowering effect on ROS production with increasing concentration from 100-500nM as compared to the positive control (SOD). When Lucigenin was used as chemiluminescence dye, 3MeEc exerted significant lowering effect with increasing concentration when compared to the positive control (SOD) whereas 4MeEc showed significant lowering effect in ROS production from 250 nM on as compared to positive control. For 4SulEc, a significant lowering effect of ROS production was only observed at 100 and 250 nM. Overall, although each metabolite shows considerable effect, 3MeEc exhibited more pronounced effect on decreasing the production of ROS as compared to other two metabolites.

Keywords: epicatechin metabolites, HO-1, Nrf2, ROS

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6895 Evaluation of DNA Oxidation and Chemical DNA Damage Using Electrochemiluminescent Enzyme/DNA Microfluidic Array

Authors: Itti Bist, Snehasis Bhakta, Di Jiang, Tia E. Keyes, Aaron Martin, Robert J. Forster, James F. Rusling

Abstract:

DNA damage from metabolites of lipophilic drugs and pollutants, generated by enzymes, represents a major toxicity pathway in humans. These metabolites can react with DNA to form either 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodG), which is the oxidative product of DNA or covalent DNA adducts, both of which are genotoxic and hence considered important biomarkers to detect cancer in humans. Therefore, detecting reactions of metabolites with DNA is an effective approach for the safety assessment of new chemicals and drugs. Here we describe a novel electrochemiluminescent (ECL) sensor array which can detect DNA oxidation and chemical DNA damage in a single array, facilitating a more accurate diagnostic tool for genotoxicity screening. Layer-by-layer assembly of DNA and enzyme are assembled on the pyrolytic graphite array which is housed in a microfluidic device for sequential detection of two type of the DNA damages. Multiple enzyme reactions are run on test compounds using the array, generating toxic metabolites in situ. These metabolites react with DNA in the films to cause DNA oxidation and chemical DNA damage which are detected by ECL generating osmium compound and ruthenium polymer, respectively. The method is further validated by the formation of 8-oxodG and DNA adduct using similar films of DNA/enzyme on magnetic bead biocolloid reactors, hydrolyzing the DNA, and analyzing by liquid chromatography-mass spectrometry (LC-MS). Hence, this combined DNA/enzyme array/LC-MS approach can efficiently explore metabolic genotoxic pathways for drugs and environmental chemicals.

Keywords: biosensor, electrochemiluminescence, DNA damage, microfluidic array

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6894 Molecular Profiling of an Oleaginous Trebouxiophycean Alga Parachlorella kessleri Subjected to Nutrient Deprivation

Authors: Pannaga Pavan Jutur

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Parachlorella kessleri, a marine unicellular green alga belonging to class Trebouxiophyceae, accumulates large amounts of oil, i.e., lipids under nutrient-deprived (-N, -P, and -S) conditions. Understanding their metabolic imprints is important for elucidating the physiological mechanisms of lipid accumulations in this microalga subjected to nutrient deprivation. Metabolic and lipidomic profiles were obtained respectively using gas chromatography-mass spectrometry (GC-MS) of P. kessleri under nutrient starvation (-N, -P and -S) conditions. Relative quantities of more than 100 metabolites were systematically compared in all these three starvation conditions. Our results demonstrate that in lipid metabolism, the quantities of neutral lipids increased significantly followed by the decrease in other metabolites involved in photosynthesis, nitrogen assimilation, etc. In conclusion, the metabolomics and lipidomic profiles have identified a few common metabolites such as citric acid, valine, and trehalose to play a significant role in the overproduction of oil by this microalga subjected to nutrient deprivation. Understanding the entire system through untargeted metabolome profiling will lead to identifying relevant metabolites involved in the biosynthesis and degradation of precursor molecules that may have the potential for biofuel production, aiming towards the vision of tomorrow’s bioenergy needs.

Keywords: algae, biofuels, nutrient stress, omics

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6893 Identification of Bioactive Metabolites from Ficus carica and Their Neuroprotective Effects of Alzheimer's Disease

Authors: Hanan Khojah, RuAngelie Edrada-Ebel

Abstract:

Neurodegenerative disease including Alzheimer’s disease is a major cause of long-term disability. Oxidative stress is frequently implicated as one of the key contributing factors to neurodegenerative diseases. Protection against neuronal damage remains a great challenge for researchers. Ficus carica (commonly known as fig) is a species of great antioxidant nutritional value comprising a protective mechanism against innumerable health disorders related to oxidative stress as well as Alzheimer’s disease. The purpose of this work was to characterize the non-polar active metabolites in Ficus carica endocarp, mesocarp, and exocarp. Crude extracts were prepared using several extraction solvents, which included 1:1 water: ethylacetate, acetone and methanol. The dried extracts were then solvent partitioned between equivalent amounts of water and ethylacetate. Purification and fractionation were accomplished by high-throughput chromatography. The isolated metabolites were tested on their effect on human neuroblastoma cell line by cell viability test and cell cytotoxicity assay with acrolein. Molecular weights of the active metabolites were determined via LC–HRESIMS and GC-EIMS. Metabolomic profiling was performed to identify the active metabolites by using differential expression analysis software (Mzmine) and SIMCA for multivariate analysis. Structural elucidation and identification of the interested active metabolites were studied by 1-D and 2-D NMR. Significant differences in bioactivity against a concentration-dependent assay on acrolein radicals were observed between the three fruit parts. However, metabolites obtained from mesocarp and the endocarp demonstrated bioactivity to scavenge ROS radical. NMR profiling demonstrated that aliphatic compounds such as γ-sitosterol tend to induce neuronal bioactivity and exhibited bioactivity on the cell viability assay. γ-Sitosterol was found in higher concentrations in the mesocarp and was considered as one of the major phytosterol in Ficus carica.

Keywords: alzheimer, Ficus carica, γ-Sitosterol, metabolomics

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6892 Implementation of Metabolomics in Conjunction with Chemometrics for the Dentification of the Differential Chemical Markers of Different Grades of Sri Lankan White, Green and Black Tea: Camellia Sinenesis L.

Authors: Dina A. Selim, Eman Shawky, Rasha M. Abu El-Khair

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In the current study, UPLC-MS/MS combined to chemometrics were applied on seven Sri Lankan tea grades; Orange Pekoe, Flowery Pekoe, Broken Orange Pekoe Fannings, Broken Orange Pekoe black tea, green tea, silver tips and golden tips white tea grades for their comprehensive metabolic profiling. Certain metabolites, namely, Theasensinin C and E, theaflavin and theacitrin appeared to be the main chemical markers of black tea type, catechin, epicatechin, epigallocatechin, methyl epigallocatechin were the main discriminatory markers of green tea type, while theanine, oolongotheanine and quercetin glycosides were the main chemical markers of white tea type. Theogalloflavin, epigallocatechin and flavonoid glycosides were the main down-accumulated metabolites while theaflavin gallate, and N-ethyl pyrrolidinone epicatechin were the chief up- accumulated metabolites between whole and broken black tea leave grades while puerin A and C and gallic acid was the main down- accumulated metabolites and N-ethyl pyrrolidinone epicatechin gallate was the main up-accumulated one between broken and fanning black tea grades.

Keywords: tea grading, Sri Lankan tea, chemometrics, metabolomics, chemical markers

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6891 Association between Levels of Volatile Organic Compound Metabolites and Cigarette Smoking-Related Urothelial Carcinoma

Authors: Chi-Jung Chung, Chao-Hsiang Chang, Chiu-Shong Liu, Sheng-Wei Li, Mu-Chi Chung, Ting-Jie Wen, Hui-Ling Lee

Abstract:

Cigarette smoke contains volatile organic compounds (VOCs), such as acrylamide, 1,3-butadiene, and benzene. This study aimed to explore the associations between the urinary levels of cotinine and VOC metabolites and the risk of urothelial carcinoma (UC). A hospital-based case–control study involving two groups matched on the basis of age ( ± 3 years) and gender was designed. UC was clinically diagnosed through urological examinations and pathologically verified. Smoking-related information was collected through questionnaires and face-to-face interviews with all study participants. Urine samples were collected for the analysis of the urinary levels of VOC metabolites, cotinine, and 8-hydroxydeoxygua- nosine (8-OHdG), which was selected as a proxy of oxidative stress. Multiple logistic regressions were applied to estimate the risk of UC. The urinary cotinine and 8-OHdG levels of the UC group were higher than those of the control group. The urinary levels of VOC metabolites, including N-acetyl-S-(2-carbamoylethyl)-L-cysteine (AAMA), N- acetyl-S-(2-carbamoyl-2-hydroxyethyl)-L-cysteine, N-acetyl-S- (4- hydroxy-2-buten-1-yl)-Lcysteine-3, trans, trans-muconic acid (t,t- MA), and S-phenylmercapturic acid (SPMA) increased as the urinary levels of cotinine increased. Relevant dose-response relationships between the risk of UC risk and the urinary levels of AAMA , t,t-MA, SPMA, and 8-OHdG were found after adjusting for potential risk factors. The UC risk of participants with high urinary levels of cotinine, AAMA, t,t-MA, SPMA, and 8-OHdG were 3.5–6-fold higher than those of other participants. Increased urinary levels of VOC metabolites were associated with smoking-related UC risk. The development of UC should be explored in large-scale in vitro or in vivo studies with the repeated measurement of VOC metabolites.

Keywords: volatile organic compound, urothelial carcinoma, cotinine, 8-hydroxydeoxyguanosine

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6890 Microbial Metabolites with Ability of Anti-Free Radicals

Authors: Yu Pu, Chien-Ping Hsiao, Chien-Chang Huang, Chieh-Lun Cheng

Abstract:

Free radicals can accelerate aging on human skin by causing lipid oxidation, protein denaturation, and even DNA mutation. Substances with the ability of anti-free radicals can be used as functional components in cosmetic products. Research are attracted to develop new anti-free radical components for cosmetic application. This study was aimed to evaluate the microbial metabolites on free radical scavenging ability. Two microorganisms, PU-01 and PU-02, were isolated from soil of hot spring environment and grew in LB agar at 50°C for 24 h. The suspension was collected by centrifugation at 4800 g for 3 min, The anti-free radical activity was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging assay. The result showed that the growth medium of PU-01 presented a higher DPPH scavenging effect than that of PU-02. This study presented potential anti-free radical components from microbial metabolites that might be applied in anti-aging cosmetics.

Keywords: anti-ageing, anti-free radical, biotechnology, microorganism

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6889 Plasmonic Nanoshells Based Metabolite Detection for in-vitro Metabolic Diagnostics and Therapeutic Evaluation

Authors: Deepanjali Gurav, Kun Qian

Abstract:

In-vitro metabolic diagnosis relies on designed materials-based analytical platforms for detection of selected metabolites in biological samples, which has a key role in disease detection and therapeutic evaluation in clinics. However, the basic challenge deals with developing a simple approach for metabolic analysis in bio-samples with high sample complexity and low molecular abundance. In this work, we report a designer plasmonic nanoshells based platform for direct detection of small metabolites in clinical samples for in-vitro metabolic diagnostics. We first synthesized a series of plasmonic core-shell particles with tunable nanoshell structures. The optimized plasmonic nanoshells as new matrices allowed fast, multiplex, sensitive, and selective LDI MS (Laser desorption/ionization mass spectrometry) detection of small metabolites in 0.5 μL of bio-fluids without enrichment or purification. Furthermore, coupling with isotopic quantification of selected metabolites, we demonstrated the use of these plasmonic nanoshells for disease detection and therapeutic evaluation in clinics. For disease detection, we identified patients with postoperative brain infection through glucose quantitation and daily monitoring by cerebrospinal fluid (CSF) analysis. For therapeutic evaluation, we investigated drug distribution in blood and CSF systems and validated the function and permeability of blood-brain/CSF-barriers, during therapeutic treatment of patients with cerebral edema for pharmacokinetic study. Our work sheds light on the design of materials for high-performance metabolic analysis and precision diagnostics in real cases.

Keywords: plasmonic nanoparticles, metabolites, fingerprinting, mass spectrometry, in-vitro diagnostics

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6888 Biological Organic or Inorganic Sulfur Sources Feeding Effects on Intake and Some Blood Metabolites of Close-Up Holstein Cows

Authors: Mehdi Kazemi-Bonchenari, Esmaeil Manidari, Vahid Keshavarz

Abstract:

This study was carried out to investigate the effects of increased level of sulfur by supplementing magnesium sulfate with or without biologically organic source in dairy cow close-up diets on dry matter intake (DMI) and some blood metabolites. The 24 multiparous close-up Holstein cows averaging body weight 687.94 kg and days until expected calving date 21.89 d were allocated in three different treatments (8 cows per each) in a completely randomized design. The first treatment (T1) has contained 0.21% sulfur (DM basis), the second treatment (T2) has contained 0.41% sulfur which entirely supplied through magnesium sulfate and the third treatment (T3) has contained 0.41% sulfur which supplied through combination of magnesium sulfate and an organic source of sulfur. All the cows were fed same diet after parturition until 21 d. The DMI for both pre-calving (P < 0.001) and post-calving was affected by treatments (P < 0.004) and T2 showed the lowest DMI among treatments. Among the blood metabolites, glucose, calcium, and copper were decreased in T2 (P < 0.05). However, blood concentrations of BHBA, NEFA, urea, CPK, and AST were increased in T2 (P < 0.05). The results of the present study indicate that although magnesium sulfate has negative effect on dairy cow health and performance, a combination of magnesium sulfate and biological organic source of sulfur in close-up diets could have positive effects on DMI and performance of Holstein dairy cows.

Keywords: organic sulfur, dairy cow, intake, blood metabolites

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6887 Heat and Humidity Induced Plastic Changes in Body Lipids and Starvation Resistance in the Tropical Zaprionus indianus of Wet-Dry Seasons

Authors: T. N. Girish, B. E. Pradeep, Ravi Parkash

Abstract:

Insects from tropical wet or dry seasons are likely to cope starvation stress through seasonal phenotypic plasticity in energy metabolites. Accordingly, we analyzed such plastic changes in Zaprionus indianus flies reared under wet or dry season-specific conditions; and also after adult acclimation at 32℃ for 1 to 6 days; and to low (40% RH) or high (70% RH) humidity. Both thermal or humidity acclimation revealed significant accumulation of body lipids for wet season flies but low humidity acclimation did not change the level of body lipids in dry season flies. Developmental and adult acclimation showed sex specific differences i.e., starvation resistance and body lipids were higher in the males of dry season but in females of wet season. We found seasonal and sex specific differences in the relative level for body lipids at death; and in the rates of accumulation or utilization of energy metabolites (body lipids, carbohydrates and proteins). Body lipids constitute the preferred energy source under starvation for flies of both the seasons. However, utilization of carbohydrates (~20% to 30%) and proteins (~20% to 25%) was evident only in dry season flies. Higher starvation resistance after thermal or humidity acclimation is achieved by increased accumulation of lipids. Adult acclimation of wet or dry season flies revealed plastic changes in mean daily fecundity despite reduction in fecundity under starvation. Thus, thermal or humidity induced plastic responses in body lipids support starvation resistance under wet or dry seasons.

Keywords: heat or humidity acclimation, plastic changes in body lipids and starvation resistance, tropical drosophilid, Wet- or Dry seasons, Zaprionus indianus

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6886 Simple Modified Method for DNA Isolation from Lyophilised Cassava Storage Roots (Manihot esculenta Crantz.)

Authors: P. K. Telengech, K. Monjero, J. Maling’a, A. Nyende, S. Gichuki

Abstract:

There is need to identify an efficient protocol for use in extraction of high quality DNA for purposes of molecular work. Cassava roots are known for their high starch content, polyphenols and other secondary metabolites which interfere with the quality of the DNA. These factors have negative interference on the various methodologies for DNA extraction. There is need to develop a simple, fast and inexpensive protocol that yields high quality DNA. In this improved Dellaporta method, the storage roots are lyophilized to reduce the water content; the extraction buffer is modified to eliminate the high polyphenols, starch and wax. This simple protocol was compared to other protocols intended for plants with similar secondary metabolites. The method gave high yield (300-950ng) and pure DNA for use in PCR analysis. This improved Dellaporta protocol allows isolation of pure DNA from starchy cassava storage roots.

Keywords: cassava storage roots, dellaporta, DNA extraction, lyophilisation, polyphenols secondary metabolites

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6885 In vitro Callus Production from Lantana Camara: A Step towards Biotransformation Studies

Authors: Maged El-Sayed Mohamed

Abstract:

Plant tissue culture practices are presented nowadays as the most promising substitute to a whole plant in the terms of secondary metabolites production. They offer the advantages of high production, tunability and they have less effect on plant ecosystems. Lantana camara is a weed, which is common all over the world as an ornamental plant. Weeds can adapt to any type of soil and climate due to their rich cellular machinery for secondary metabolites’ production. This characteristic is found in Lantana camara as a plant of very rich diversity of secondary metabolites with no dominant class of compounds. Aim: This trait has encouraged the author to develop tissue culture experiments for Lantana camara to be a platform for production and manipulation of secondary metabolites through biotransformation. Methodology: The plant was collected in its flowering stage in September 2014, from which explants were prepared from shoot tip, auxiliary bud and leaf. Different types of culture media were tried as well as four phytohormones and their combinations; NAA, 2,4-D, BAP and kinetin. Explants were grown in dark or in 12 hours dark and light cycles at 25°C. A metabolic profile for the produced callus was made and then compared to the whole plant profile. The metabolic profile was made using GC-MS for volatile constituents (extracted by n-hexane) and by HPLC-MS and capillary electrophoresis-mass spectrometry (CE-MS) for non-volatile constituents (extracted by ethanol and water). Results: The best conditions for the callus induction was achieved using MS media supplied with 30 gm sucrose and NAA/BAP (1:0.2 mg/L). Initiation of callus was favoured by incubation in dark for 20 day. The callus produced under these conditions showed yellow colour, which changed to brownish after 30 days. The rate of callus growth was high, expressed in the callus diameter, which reached to 1.15±0.2 cm in 30 days; however, the induction of callus delayed for 15 days. The metabolic profile for both volatile and non-volatile constituents of callus showed more simple background metabolites than the whole plant with two new (unresolved) peaks in the callus’ nonvolatile constituents’ chromatogram. Conclusion: Lantana camara callus production can be itself a source of new secondary metabolites and could be used for biotransformation studies due to its simple metabolic background, which allow easy identification of newly formed metabolites. The callus production gathered the simple metabolic background with the rich cellular secondary metabolite machinery of the plant, which could be elicited to produce valuable medicinally active products.

Keywords: capillary electrophoresis-mass spectrometry, gas chromatography, metabolic profile, plant tissue culture

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6884 Changes in Amounts of Glycyrrhizin and Phenolic Compounds of Glycrrhiza glabra L. Seedlings Treated by Copper and Zinc Oxide

Authors: Roya Razavizadeh, Razieh Soltaninejad, Hakimeh Oloumi

Abstract:

Glycyrrhiza glabra L. (Licorice) is one of the oldest medicinal plants in Iran and secondary metabolites present in the plant root is used in food and pharmaceutical industries. With the use of heavy metals as elicitors, plant secondary metabolite production can be increased. In this study, the effects of the concentrations of 1 and 10 μM of zinc oxide and copper oxide on the contents of reducing sugars (as precursor of secondary metabolites), proline, glycyrrhizin, total phenolic compounds, flavonoids and anthocyanin in Glycyrrhiza glabra seedlings were investigated. Also, the correlation between the content of these metabolites in the treated seedlings was examined using Pearson's test. The amount of reducing sugars at concentration of 10 μM zinc oxide was decreased. Whereas, the amounts of proline and glycyrrhizin under treatment 1 and 10 μM copper oxide and 1 μM zinc oxide compared with the control plants was increased. The content of total phenolic compounds was increased with increasing concentrations of copper oxide. The highest amount of flavonoids was observed at concentrations of 1 and 10 μM copper oxide. Anthocyanin content was increased in concentration of 1 μM copper oxide. Also, the tannin content of the Glycyrrhiza glabra seedlings at concentrations of 10 μM zinc oxide was increased. Based on the result it seemed that at concentrations of 1 and 10 μM copper oxide the amount of glycyrrhizin, phenolic compounds, flavonoids, anthocyanins were significantly increased, whereas, zinc oxide had no significant impact on the levels of these metabolites.

Keywords: zinc oxide, copper oxide, phenolic compounds, licorice (glycyrrhiza glabra L.), glycyrrhizin

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6883 Human Lens Metabolome: A Combined LC-MS and NMR Study

Authors: Vadim V. Yanshole, Lyudmila V. Yanshole, Alexey S. Kiryutin, Timofey D. Verkhovod, Yuri P. Tsentalovich

Abstract:

Cataract, or clouding of the eye lens, is the leading cause of vision impairment in the world. The lens tissue have very specific structure: It does not have vascular system, the lens proteins – crystallins – do not turnover throughout lifespan. The protection of lens proteins is provided by the metabolites which diffuse inside the lens from the aqueous humor or synthesized in the lens epithelial layer. Therefore, the study of changes in the metabolite composition of a cataractous lens as compared to a normal lens may elucidate the possible mechanisms of the cataract formation. Quantitative metabolomic profiles of normal and cataractous human lenses were obtained with the combined use of high-frequency nuclear magnetic resonance (NMR) and ion-pairing high-performance liquid chromatography with high-resolution mass-spectrometric detection (LC-MS) methods. The quantitative content of more than fifty metabolites has been determined in this work for normal aged and cataractous human lenses. The most abundant metabolites in the normal lens are myo-inositol, lactate, creatine, glutathione, glutamate, and glucose. For the majority of metabolites, their levels in the lens cortex and nucleus are similar, with the few exceptions including antioxidants and UV filters: The concentrations of glutathione, ascorbate and NAD in the lens nucleus decrease as compared to the cortex, while the levels of the secondary UV filters formed from primary UV filters in redox processes increase. That confirms that the lens core is metabolically inert, and the metabolic activity in the lens nucleus is mostly restricted by protection from the oxidative stress caused by UV irradiation, UV filter spontaneous decomposition, or other factors. It was found that the metabolomic composition of normal and age-matched cataractous human lenses differ significantly. The content of the most important metabolites – antioxidants, UV filters, and osmolytes – in the cataractous nucleus is at least ten fold lower than in the normal nucleus. One may suppose that the majority of these metabolites are synthesized in the lens epithelial layer, and that age-related cataractogenesis might originate from the dysfunction of the lens epithelial cells. Comprehensive quantitative metabolic profiles of the human eye lens have been acquired for the first time. The obtained data can be used for the analysis of changes in the lens chemical composition occurring with age and with the cataract development.

Keywords: cataract, lens, NMR, LC-MS, metabolome

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6882 Transcriptomic Analysis for Differential Expression of Genes Involved in Secondary Metabolite Production in Narcissus Bulb and in vitro Callus

Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls

Abstract:

The Amaryllidaceae genus Narcissus contains secondary metabolites, which are important sources of bioactive compounds such as pharmaceuticals indicating that their biological activity extends from the native plant to humans. Transcriptome analysis (RNA-seq) is an effective platform for the identification and functional characterization of candidate genes as well as to identify genes encoding uncharacterized enzymes. The biotechnological production of secondary metabolites in plant cell or organ cultures has become a tempting alternative to the extraction of whole plant material. The biochemical pathways for the production of secondary metabolites require primary metabolites to undergo a series of modifications catalyzed by enzymes such as cytochrome P450s, methyltransferases, glycosyltransferases, and acyltransferases. Differential gene expression analysis of Narcissus was obtained from two conditions, i.e. field and in vitro callus. Callus was obtained from modified MS (Murashige and Skoog) media supplemented with growth regulators and twin-scale explants from Narcissus cv. Carlton bulb. A total of 2153 differentially expressed transcripts were detected in Narcissus bulb and in vitro callus, and 78.95% of those were annotated. It showed the expression of genes involved in the biosynthesis of alkaloids were present in both conditions i.e. cytochrome P450s, O-methyltransferase (OMTs), NADP/NADPH dehydrogenases or reductases, SAM-synthetases or decarboxylases, 3-ketoacyl-CoA, acyl-CoA, cinnamoyl-CoA, cinnamate 4-hydroxylase, alcohol dehydrogenase, caffeic acid, N-methyltransferase, and NADPH-cytochrome P450s. However, cytochrome P450s and OMTs involved in the later stage of Amaryllidaceae alkaloids biosynthesis were mainly up-regulated in field samples. Whereas, the enzymes involved in initial biosynthetic pathways i.e. fructose biphosphate adolase, aminotransferases, dehydrogenases, hydroxyl methyl glutarate and glutamate synthase leading to the biosynthesis of precursors; tyrosine, phenylalanine and tryptophan for secondary metabolites were up-regulated in callus. The knowledge of probable genes involved in secondary metabolism and their regulation in different tissues will provide insight into the Narcissus plant biology related to alkaloid production.

Keywords: narcissus, callus, transcriptomics, secondary metabolites

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6881 Simultaneous Production of Forskolin and Rosmarinic Acid in vitro Cultures of Coleus Forskohlii Briq

Authors: Ennus Tajuddin Tamboli, Madhukar Garg, Mohd. Mujeeb, Sayeed Ahmad

Abstract:

An efficient protocol for simultaneous production of forskolin and rosmarinic acid in in vitro callus derived from the leaves of Coleus forskohlii Briq. has been developed. MS media was used for the establishment of cultures and NAA + 6-BA (1.0 ppm) was found best for callus growth. The callus was further subjected to treatment with various elicitor/precursors viz. chitosan, thidiazuron and methyl jasmonate to observe their effect on production of biomass and accumulation of secondary metabolites. The content of forskolin and rosmarinic acid were estimated by HPTLC, in comparison to natural explant which showed 2 fold and 10 fold rise in forskolin and rosmarinic acid content, respectively. Methy1 jasmonate 50 µM was found best for production of forskolin, whereas thidiazuron showed best results in the yield of rosmarinic acid, separately in static culture. However, combined treatment in suspension culture showed moderated effect for increase in secondary metabolites but the biomass increased significantly as compared to static culture.

Keywords: plant tissue culture, secondary metabolites, coleus, forskolin, rosmarinic acid, HPTLC

Procedia PDF Downloads 258