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1.
Brassinosteroids (BRs) are the polyhydroxylated plant hormones sharing a common resemblance with animal steroids. They are active even at very low concentrations and are implicated for their pleiotropic involvement in diverse physiological processes and defense strategies during stress in plants. These compounds are well apparent in the plant kingdom with higher amounts in juvenile tissues. A total of 62 steroidal compounds have been identified so far. Keeping their significance in mind, researchers not only have worked extensively on their isolation, but also they were synthesizing their synthetic isomers. Different analytical techniques like HPLC, GC-MS, LC-MS/MS, UPLC-MS/MS, and bioassay-based methods have been used for their isolation, detection, and characterization from composite plant materials. Therefore, this review provides comprehensive information to the readers intending to isolate and characterize BRs, using either laborious techniques or modern-day more efficient methods. 相似文献
2.
Extreme natural habitats like halophytes, marsh land, and marine environment are suitable arena for chemical ecology between plants and microbes having environmental impact. Endophytes are an ecofriendly option for the promotion of plant growth and to serve as sustainable resource of novel bioactive natural products. The present study, focusing on biodiversity of bacterial endophytes from Salicornia brachiata, led to isolation of around 336 bacterial endophytes. Phylogenetic analysis of 63 endophytes revealed 13 genera with 27 different species, belonging to 3 major groups: Firmicutes, Proteobacteria, and Actinobacteria. 30% endophytic isolates belonging to various genera demonstrated broad-spectrum antibacterial and antifungal activities against a panel of human, plant, and aquatic infectious agents. An endophytic isolate Bacillus amyloliquefaciens 5NPA-1, exhibited strong in-vitro antibacterial activity against human pathogen Staphylococcus aureus and phytopathogen Xanthomonas campestris. Investigation through LC–MS/MS-based molecular networking and bioactivity-guided purification led to the identification of three bioactive compounds belonging to lipopeptide class based on 1H-, 13C-NMR and MS analysis. To our knowledge, this is the first report studying bacterial endophytic biodiversity of Salicornia brachiata and the isolation of bioactive compounds from its endophyte. Overall, the present study provides insights into the diversity of endophytes associated with the plants from the extreme environment as a rich source of metabolites with remarkable agricultural applications and therapeutic properties. 相似文献
3.
A method has been developed for the quantitative isolation of glucosinolates by ion-exchange chromatography and high voltage electrophoresis avoiding strongly alkaline and acidic conditions. The compounds were identified by 1H and 13C NMR spectroscopy and through the products arising from enzymatic, acid and alkaline hydrolysis. The method is well suited for the isolation and identification of glucosinolates containing aglucone parts which produce non-volatile compounds on enzymatic hydrolysis. The method has been used in the isolation and identification of 2-hydroxy-2-methylpropylglucosinolate from Reseda alba, 2-hydroxy-2-phenylethylglucosinolate from R. luteola and a new glucosinolate, o-(α-l-rhamnopyranosyloxy)benzylglucosinolate, occurring in R. odorata. The glucosinolate content in different parts of this plant has been determined and the metabolism of glucosinolates is briefly discussed. 相似文献
4.
Current research in drug discovery from medicinal plants involves a multifaceted approach combining botanical, phytochemical, biological, and molecular techniques. Medicinal plant drug discovery continues to provide new and important leads against various pharmacological targets including cancer, HIV/AIDS, Alzheimer's, malaria, and pain. Several natural product drugs of plant origin have either recently been introduced to the United States market, including arteether, galantamine, nitisinone, and tiotropium, or are currently involved in late-phase clinical trials. As part of our National Cooperative Drug Discovery Group (NCDDG) research project, numerous compounds from tropical rainforest plant species with potential anticancer activity have been identified. Our group has also isolated several compounds, mainly from edible plant species or plants used as dietary supplements, that may act as chemopreventive agents. Although drug discovery from medicinal plants continues to provide an important source of new drug leads, numerous challenges are encountered including the procurement of plant materials, the selection and implementation of appropriate high-throughput screening bioassays, and the scale-up of active compounds. 相似文献
5.
The combination of the hyphenated techniques LC–MS and LC–SPE–NMR constitutes a powerful platform for the rapid isolation and identification of minor components from natural sources. Electronic circular dichroism (ECD) is a useful tool to determine the absolute configuration of small quantities of chiral molecules. In order to search for minor constituents present in an Ormocarpum kirkii extract, these techniques were applied for the separation and structure elucidation of a series of isoflavanones, biflavanones and biscoumarins. After optimization of chromatographic conditions and subsequent isolation, MS and 1D and 2D NMR data were collected. Experimental and calculated ECD spectra were used in conjunction with NMR data to confirm the absolute configuration of these compounds. Eight compounds were identified for the first time and six have been previously reported. The present approach offers a strategy for accelerating research on natural products. 相似文献
6.
Chromatography is the lynchpin of phytochemistry and is the key to obtaining pure compounds for structure elucidation, for pharmacological testing or for development into therapeuticals. It also plays a fundamental role as an analytical technique for quality control and standardisation of phytotherapeuticals. Although liquid chromatography is barely 100 years old, an extraordinary variety of instrumental and ancillary equipment is available, notably in the domain of high-performance liquid chromatography. It is impossible to touch all areas of chromatography in such a review but certain areas are worthy of mention: HPLC, HPTLC, UPLC and countercurrent chromatography. Another important addition has been the development of hyphenated techniques involving HPLC: LC/UV, LC/MS, LC/MS(n) and LC/NMR. These are indispensable nowadays for the early detection and identification of compounds in crude plant extracts. 相似文献
7.
Several new plant breeding techniques (NPBTs) have been developed during the last decade, and make it possible to precisely perform genome modifications in plants. The major problem, other than technical aspects, is the vagueness of regulation concerning these new techniques. Since the definition of eight NPBTs by a European expert group in 2007, there has been an ongoing debate on whether the resulting plants and their products are covered by GMO legislation. Obviously, cover by GMO legislation would severely hamper the use of NPBT, because genetically modified plants must pass a costly and time‐consuming GMO approval procedure in the EU. In this review, we compare some of the NPBTs defined by the EU expert group with classical breeding techniques and conventional transgenic plants. The list of NPBTs may be shortened (or extended) during the international discussion process initiated by the Organization for Economic Co‐operation and Development. From the scientific point of view, it may be argued that plants developed by NPBTs are often indistinguishable from classically bred plants and are not expected to possess higher risks for health and the environment. In light of the debate on the future regulation of NPBTs and the accumulated evidence on the biosafety of genetically modified plants that have been commercialized and risk‐assessed worldwide, it may be suggested that plants modified by crop genetic improvement technologies, including genetic modification, NPBTs or other future techniques, should be evaluated according to the new trait and the resulting end product rather than the technique used to create the new plant variety. 相似文献
8.
随着对药用植物内生真菌研究的深入,从药用植物内生真菌中寻找新的生物活性成分已成为研究热点。内生真菌对药用植物的生长及活性成分的形成都有影响,内生真菌活性成分已成为发现新颖结构化合物及新药物的潜在资源。简要综述了近年来在内生真菌的分离鉴定、发酵、次生代谢产物、与宿主的关系等方面的研究进展。 相似文献
9.
The most important sweet substance known is sucrose, which is obtained commercially from sugar cane and sugar beet. Because the intake of sucrose has been associated with a number of adverse effects on health, an intensive search has been undertaken to find alternative substances to satisfy the human craving for a sweet taste. Many other plant‐derived compounds are sweet, ranging in structural complexity from sugars and polyhydric alcohols through diterpene and triterpene glycosides to proteins; some of these compounds are intensely sweet, being hundreds or even thousand times sweeter than sucrose, and offer potential for commercial use in dietetic and diabetic foodstuffs. The present review examines the role of ethnobotany in the discovery of sweet‐tasting plants, the chemical isolation and elucidation of the sweet compounds, and some safety and sensory evaluation aspects of these compounds. A discussion on the future prospects of discovering and developing new plant‐derived sweeteners concludes the review. 相似文献
10.
The bulk of indole‐3‐acetic acid (IAA) in plants is found in the form of conjugated molecules, yet past research on identifying these compounds has largely relied on methods that were both laborious and inefficient. Using recent advances in analytical instrumentation, we have developed a simple yet powerful liquid chromatography–mass spectrometry (LC–MS)‐based method for the facile characterization of the small IAA conjugate profile of plants. The method uses the well‐known quinolinium ion ( m/z 130.0651) generated in MS processes as a signature with high mass accuracy that can be used to screen plant extracts for indolic compounds, including IAA conjugates. We reinvestigated Glycine max (soybean) for its indoles and found indole‐3‐acetyl‐trytophan (IA‐Trp) in addition to the already known indole‐3‐acetyl‐aspartic acid (IA‐Asp) and indole‐3‐acetyl‐glutamic acid (IA‐Glu) conjugates. Surprisingly, several organic acid conjugates of tryptophan were also discovered, many of which have not been reported in planta before. These compounds may have important physiological roles in tryptophan metabolism, which in turn can affect human nutrition. We also demonstrated the general applicability of this method by identifying indolic compounds in different plant tissues of diverse phylogenetic origins. It involves minimal sample preparation but can work in conjunction with sample enrichment techniques. This method enables quick screening of IAA conjugates in both previously characterized as well as uncharacterized species, and facilitates the identification of indolic compounds in general. 相似文献
11.
Phytochemical investigation of the methanol extract of the root bark of Dichostachys cinerea (Fabaceae) led to the isolation and characterization of 3 new pyrrolidine-derived compounds with 12 carbon side chain and their glycosides. The structures of these compounds ( 1–3) were established using spectroscopic analytical methods comprising NMR experiments, ESI and HR-ESIMS mass spectrometry, and mass tandem spectrometry. The compounds are related to some polyhydroxypyrrolidine type-iminosugars (broussonetinine and broussonetine) found in the restrictive plant families Moraceae, Campanulaceae and Hyacinthaceae, but with, no hydroxyl group on the pyrrolidine ring. Pyrrolidines with long-side chains have been found in the plant family, Araceae ( Arisarum vulgare). The presence of compounds 1– 3 in D. cinerea expands the range of distribution of pyrrolidine and its derivatives in plants, and thus presenting a new addition to the molecular diversity of pyrrolidine which could explain some of pharmacological properties of sourced plants. 相似文献
13.
Natural products chemistry has witnessed many new developments in the last 5 years like extractions with subcritical water
and ionic liquids, LC/HRMS and LC/SPE/cryo-NMR, UHPLC, TLC/MS, MS-based preparative HPLC, comprehensive chromatography (GC × GC,
LC × LC), high-throughput screening, introduction of monolithic columns, miniaturisation, and automated structure identification.
Nevertheless identifying bioactive constituents in complex plant extracts remains a tedious process. The classical approach
of bioassay guided fractionation is time-consuming while off-line screening of extracts does not provide information on individual
compounds and sometimes suffers from false positives or negatives. One way out of this is by coupling chromatography with
chemical or biochemical assays, so called high resolution screening. An example is the development of HPLC on-line assays
for antioxidants. By the post-column addition of a relatively stable coloured radical like DPPH • or ABTS •+, radical scavengers are detected as negative peaks because in a reaction coil they reduce the model radical to its reduced,
non-coloured form. When combined with LC/DAD/MS and LC/SPE/NMR, reliable identification of active constituents becomes possible
without the necessity of ever isolating them in a classical sense. Also for finding leads for new drugs, combining HPLC with
biochemical assays is interesting but technically more difficult. Most enzymes do not work at the organic modifier concentrations
commonly encountered in RP-HPLC and the reaction time is often longer requiring dilution and lengthy coils respectively. Therefore,
new techniques have to be implemented to gain the required sensitivity for on-line enzyme assays. For stable analytes, high
temperature LC offers a solution to the organic modifier problem. When enzymes are highly expensive, like those used in the
screening for Cytochrome P450 inhibitors, miniaturisation to chip format may offer a way out. Microreactors (chips) are not
only useful for miniaturising larger assays but also offer completely new prospects in phytochemical analysis. One such application
is in the sample clean-up of acids and bases like alkaloids. In a lay-out of three parallel channels of 100 μm width with
the middle one containing organic phase and the two outer ones water of high pH (feed phase) and low pH (trapping phase) such
a chip replaces two classical LLE steps but is much faster and requires less solvents and less manpower input. 相似文献
14.
One of the major challenges facing the pharmaceutical industry today is finding new ways to increase productivity, decrease costs whilst still ultimately developing new therapies that enhance human health. To help address these challenges the utilisation of analytical technologies and high-throughput automated platforms has been employed; in order to perform more experiments in a shorter time frame with increased data quality. One of the main in vitro techniques to assess new chemical entities in a discovery setting has been the use of recombinant liver enzymes, microsomes and hepatocytes. These techniques can help predict in vivo metabolism, clearance and potential drug–drug interactions of these new compounds by cytochrome P450s (the major drug metabolising enzymes). This in vitro methodology has been totally transformed in recent times by the use of automated liquid handling and HPLC tandem mass spectrometry detection techniques (LC–MS/MS). This review aims looking at recent advances in the methodology used to investigate drug metabolism by cytochrome P450s; including an up to date summary of high-throughput platforms including the use of automation and LC–MS/MS to facilitate greater throughput, chromatographic resolution and data quality. 相似文献
15.
Nature has developed an outstanding bio- and molecular diversity as a result of billion years of evolution resulting in a tremendous number of secondary metabolites. Among them a small part has been so far explored for discovery of lead compounds. The lead discovery from natural sources is a technological challenge for the pharmaceutical industry. The bio- and molecular diversity in plants, animals and microorganisms, as sources for new leads, and the remarkable recent developments in NMR, mass spectrometry coupled with advanced separation techniques (LC and GC), high throughput screening, and structure-based virtual screening are discussed in this article. 相似文献
16.
A phytochemical study on the most bioactive extract from Tectona grandis led to the isolation of two new norlignans, tectonoelin A and tectonoelin B, together with ten known compounds. The structures of the compounds were determined by a combination of 1D and 2D NMR techniques. This is the first time that this type of compound (C8–C8′ linkage norlignans) has been isolated from a dicotyledon. The general bioactivities of the isolated compounds have been studied using etiolated wheat coleoptiles. The activities showed that the isolated lignans and norlignans should be part of the defence mechanisms of this plant. 相似文献
17.
As a part of a project designed to investigate Colchicum species in Jordan, the chemical constituents of Colchicum crocifolium Boiss. (Colchicaceae) were investigated using LC‐MS and LC–UV/Vis PDA. A decision tree for working with colchicinods has been developed by incorporating data from LC‐UV/PDA and LC‐MS. This dereplication strategy draws upon the UV/PDA spectra to classify compounds into one of four structural groups and combines this with retention time and mass spectra/molecular weight to identify the compounds. This strategy was applied on a small amount of extract (2 mg) of Colchicum crocifolium to dereplicate 10 known compounds from four different structural groups, namely (?)‐demecolcine, 2‐demethyl‐(?)‐colchicine or3‐demethyl‐(?)‐colchicine, N‐deacetyl‐(?)‐colchicine, (?)‐colchiciline, (?)‐colchicine, β‐lumidemecolcine, 2‐demethyl‐ β‐lumicolchicine or 3‐demethyl‐ β‐lumicolchicine, N, N‐dimethyl ‐N‐deacetyl‐ β‐lumicornigerine, (?)‐isoandrocymbine and (?)‐autumnaline. Furthermore, a new compound was identi?ed as N, N‐dimethyl ‐N‐deacetyl‐(?)‐cornigerine. Three compounds, which had molecular ions at m/ z 325, 340 and 374, could not be dereplicated into any obvious structural classes that have been isolated in our laboratories previously or reported in the literature. Copyright © 2008 John Wiley & Sons, Ltd. 相似文献
18.
Quantification of fatty acids has been crucial to elucidate lipid biosynthesis pathways in plants. To date, fatty acid identification and quantification has relied mainly on gas chromatography (GC) coupled to flame ionization detection (FID) or mass spectrometry (MS), which requires the derivatization of samples and the use of chemical standards for annotation. Here we present an alternative method based on a simple procedure for the hydrolysis of lipids, so that fatty acids can be quantified by liquid chromatography mass spectrometry (LC‐MS) analysis. Proper peak annotation of the fatty acids in the LC‐MS‐based methods has been achieved by LC‐MS measurements of authentic standard compounds and elemental formula annotation supported by 13C isotope‐labeled Arabidopsis. As a proof of concept, we have compared the analysis by LC‐MS and GC‐FID of two previously characterized Arabidopsis thaliana knock‐out mutants for FAD6 and FAD7 desaturase genes. These results are discussed in light of lipidomic profiles obtained from the same samples. In addition, we performed untargeted LC‐MS analysis to determine the fatty acid content of two diatom species. Our results indicate that both LC‐MS and GC‐FID analyses are comparable, but that because of higher sensitivity and selectivity the LC‐MS‐based method allows for a broader coverage and determination of novel fatty acids. 相似文献
19.
Coastal plants live in heterogeneous and potentially stressful environments in which multiple stress factors may coexist. Some of these constraints can induce oxidative stress with consequent damage to cell components and structures. To contrast oxidative damage plants have evolved antioxidant systems, including both enzymatic and non-enzymatic molecules. The aim of this study was to highlight main physiological traits evolved by plants to survive in coastal environment through a comparison of nutritional and physiological parameters between dune (DC) and laboratory-grown (LC) plants of Calystegia soldanella (L.), a typical dune plant. In comparison with laboratory plants, dune plants living on a soil with relatively low nutrient content, were characterised by lower total nitrogen, K + and phosphate content and by lower K +/Na +, PO 4 2?/Cl ? and N/Cl ? ratios. Pigment content was significantly higher in LC than in DC plants. Despite their higher hydrogen peroxide content and lipid peroxidation, dune plants had a membrane damage, assessed by the electrolytic conductivity method, not significantly different from that of LC plants. Phenol and ascorbate pools, glutathione reductase and catalase activities were significantly higher in dune than in laboratory plants. Although the stress level was high, coastal plants were well protected against oxidative damage and proline, phenols, ascorbate, glutathione reductase and catalase seemed to play a pivotal role in plant adaptation to the constraints of coastal environment. 相似文献
20.
As part of our search of new bioactive compounds from indigenous medicinal plants, phytochemical investigation of the aerial parts of Fagonia indica Burm led to the isolation of seven compounds including two new compounds, namely, indicacin ( 1) and fagonicin ( 2), and five known compounds ( 3– 7) from the methanol extract. Compounds 6 and 7 are hitherto unreported from this plant. The structures of the new compounds were elucidated from their spectral data, mainly HREIMS, 1D NMR ( 1H, 13C NMR, and DEPT) and 2D NMR (COSY, NOESY, HSQC and HMBC), and by comparison with the literature data. The new compounds 1 and 2 were assayed for their cytotoxicity against human colorectal cancer cell line H-29. Compound 1 exhibited 51.40% cytotoxicity at 6.25 μM/mL dose whereas compound 2 demonstrated 39.3% cytotoxicity at the same dose. 相似文献
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