Phytochemical Analysis

Current research reports and chronological list of recent articles.


The international scientific journal Phytochemical Analysis is devoted to the publication of original articles concerning the development, improvement, validation and/or extension of application of analytical methodology in the plant sciences. The spectrum of coverage is broad, encompassing methods and techniques relevant to the detection (including bio-screening), extraction, separation, purification, identification and quantification of compounds in plant biochemistry, plant cellular and molecular biology, plant biotechnology, the food sciences, agriculture and horticulture

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Additional research articles see Current Chemistry Research Articles. Magazines with similar content (phytochemistry):

 - Phytochemistry.

 - Phytochemistry Letters.

 - Phytochemistry Reviews.



Phytochemical Analysis - Abstracts



Utilization of the Ability to Induce Activation of the Nuclear Factor (Erythroid-derived 2)-like Factor 2 (Nrf2) to Assess Potential Cancer Chemopreventive Activity of Liquorice Samples

Introduction Nuclear factor (erythroid-derived 2)-like factor 2 (Nrf2) is a transcription factor that regulates expression of many detoxification enzymes. Nrf2-antioxidant responsive element (Nrf2-ARE) signalling pathway can be a target for cancer chemoprevention. Glycyrrhiza glabra, common name, ‘liquorice’, is used as a sweetening and flavouring agent, and traditionally, to treat various ailments, and implicated to chemoprevention. However, its chemopreventive property has not yet been scientifically substantiated. Objective To assess the ability of liquorice root samples to induce Nrf2 activation correlating to their potential chemopreventive property. Methods The ability of nine methanolic extracts of liquorice root samples, collected from various geographical origins, to induce Nrf2 activation was determined by the luciferase reporter assay using the ARE-reporter cell line, AREc32. The antioxidant properties were determined by the 2,2-diphenyl-1-picryhydrazyl (DPPH) and the ferric-reducing antioxidant power (FRAP) assays. Results All extracts exhibited free-radical-scavenging property (RC50 = 136.39–635.66 µg/mL). The reducing capacity of ferrous ion was 214.46–465.59 μM Fe(II)/g. Nrf2 activation indicated that all extracts induced expression of ARE-driven luciferase activity with a maximum induction of 2.3 fold relative to control. These activities varied for samples from one geographical location to another. Conclusions The present findings add to the existing knowledge of cancer chemoprevention by plant-derived extracts or purified phytochemicals, particularly the potential use of liquorice for this purpose. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 16.08.2016


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No abstract is available for this article.
Datum: 16.08.2016


Issue Information

No abstract is available for this article.
Datum: 16.08.2016


An Enzyme-linked Immunosorbent Assay for Genistein 7-O-[α-rhamnopyranosyl-(16)]-β-glucopyranoside Determination in Derris scandens using a Polyclonal Antibody

Introduction Genistein 7-O-[α-rhamnopyranosyl-(16)]-β-glucopyranoside (GTG) is a major bioactive compound in Derris scandens. It is responsible for anti-inflammatory activity by inhibition of cyclooxygenase and lipoxygenase. There are many commercial products of D. scandens available in Thailand. Objective To develop an enzyme-linked immunosorbent assay (ELISA) for the quantitative analysis of GTG in plant material and derived products using a polyclonal antibody. Methods An immunogen was synthesised by conjugating GTG with a carrier protein. The polyclonal antibody against GTG (GTG-PAb) was produced in New Zealand white rabbits. The ELISA method was validated for specificity, sensitivity, accuracy, precision and correlation with HPLC. Results The polyclonal antibody was specific to GTG and genistin within the range of compounds tested. The GTG ELISA was applied in the range 0.04–10.00 μg/mL with a limit of detection of 0.03 μg/mL. The recovery of GTG in spiked Derris scandens extracts ranged from 100.7 to 107.0%, with a coefficient of variation less than 7.0%. The intra- and inter-assay variations were less than 5.0%. The ELISA showed a good correlation with HPLC-UV analysis for GTG determination in samples, with a coefficient of determination (r2) of 0.9880. Conclusion An ELISA was established for GTG determination in Derris scandens. The GTG-PAb can react with GTG and genistin, but genistin has not been found in the plant. Therefore, the ELISA can be used for high throughput quality control of GTG content in D. scandens and its products. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


Metabolomic Fingerprinting of Romaneschi Globe Artichokes by NMR Spectroscopy and Multivariate Data Analysis

Introduction Globe artichoke (Cynara cardunculus L. var. scolymus L. Fiori) and cardoon (Cynara cardunculus L. var. altilis DC) are sources of nutraceuticals and bioactive compounds. Objectives To apply a NMR metabolomic fingerprinting approach to Cynara cardunculus heads to obtain simultaneous identification and quantitation of the major classes of organic compounds. Methodology The edible part of 14 Globe artichoke populations, belonging to the Romaneschi varietal group, were extracted to obtain apolar and polar organic extracts. The analysis was also extended to one species of cultivated cardoon for comparison. The 1H-NMR of the extracts allowed simultaneous identification of the bioactive metabolites whose quantitation have been obtained by spectral integration followed by principal component analysis (PCA). Results Apolar organic extracts were mainly based on highly unsaturated long chain lipids. Polar organic extracts contained organic acids, amino acids, sugars (mainly inulin), caffeoyl derivatives (mainly cynarin), flavonoids, and terpenes. The level of nutraceuticals was found to be highest in the Italian landraces Bianco di Pertosa zia E and Natalina while cardoon showed the lowest content of all metabolites thus confirming the genetic distance between artichokes and cardoon. Conclusion Metabolomic approach coupling NMR spectroscopy with multivariate data analysis allowed for a detailed metabolite profile of artichoke and cardoon varieties to be obtained. Relevant differences in the relative content of the metabolites were observed for the species analysed. This work is the first application of 1H-NMR with multivariate statistics to provide a metabolomic fingerprinting of Cynara scolymus. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


Simultaneous Determination of Bioactive Monoterpene Indole Alkaloids in Ethanolic Extract of Seven Rauvolfia Species using UHPLC with Hybrid Triple Quadrupole Linear Ion Trap Mass Spectrometry

Introduction Rauvolfia serpentina is an endangered plant species due to its over-exploitation. It has highly commercial and economic importance due to the presence of bioactive monoterpene indole alkaloids (MIAs) such as ajmaline, yohimbine, ajmalicine, serpentine and reserpine. Objective To develop a validated, rapid, sensitive and selective ultra-high-performance liquid chromatography coupled with hybrid triple quadrupole-linear ion trap mass spectrometry (UHPLC-QqQLIT-MS/MS) method in the multiple reaction monitoring (MRM) mode for simultaneous determination of bioactive MIAs in ethanolic extract of seven Rauvolfia species and herbal formulations. Methods The separation of MIAs was achieved on an ACQUITY UPLC BEH™ C18 column (1.7 μm, 2.1 mm × 50 mm) using a gradient mobile phase (0.1% aqueous formic acid and acetonitrile) at flow rate 0.3 μL/min in 7 min. The validated method showed good linearity (r2 ≥ 0.9999), limit of detection (LOD) (0.06–0.15 ng/mL), limit of quantitation (LOQ) (0.18–0.44 ng/mL), precisions [intraday: relative standard deviation (RSD) ≤ 2.24%, interday: RSD ≤ 2.74%], stability (RSD ≤ 1.53%) and overall recovery (RSD ≤ 2.23%). Results The validated method was applied to quantitate MIAs. Root of Rauvolfia vomitoria showed a high content of ajmaline (48.43 mg/g), serpentine (87.77 mg/g) whereas high quantities of yohimbine (100.21 mg/g) and ajmalicine (120.51 mg/g) were detected in R. tetraphylla. High content of reserpine was detected in R. micrantha (35.18 mg/g) and R. serpentina (32.38 mg/g). Conclusion The encouraging results of this study may lead to easy selection of suitable Rauvolfia species according to the abundance of MIAs. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


GC-MS and q-NMR based chemotaxonomic evaluation of two Leonurus species

Introduction The genus Leonurus L. (fam: Lamiaceae) is represented in Uzbekistan by two species, L. panzerioides Popov. and L. turkestanicus V. I. Krecz. & Kuprian, which are used to treat nervous disorders and also as sedative and hypotensive agents. Objectives To establish the taxonomic status of Leonurus panzerioides and L. turkestanicus based on their chemical constituents analysed by GC–MS and q-NMR. Materials and Methods Quantitative 1H-NMR (q-NMR) was used to identify and quantify known major components in the methanol extracts of these two species. Additionally, the chemical composition of the essential oils obtained from the aerial parts of these plants were analysed by GC–MS. Results The q-NMR analyses of Leonurus panzerioides and L. turkestanicus revealed the presence of 8-acetylharpagide, harpagide, leonurine and stachydrine as major components. Using the GC–MS method, overall 24 and 39 constituents were identified, respectively, from L. panzerioides and L. turkestanicus oils. The major constituents of the essential oil of L. panzerioides were eugenol (30.9%) and p-vinyl guaiacol (15.8%), whereas thymol (40.1%) and octen-3-ol (13.1%) were the principal compounds in the essential oil of L. turkestanicus. Conclusion The major components in Leonurus panzerioides and L. turkestanicus as identified by the GC–MS and q-NMR analyses, were similar to those present in other Leonurus species and thus provided chemotaxonomic evidence for the placement of these species under the genus Leonurus. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


Supercritical Extraction of Scopoletin from Helichrysum italicum (Roth) G. Don Flowers

Introduction The increasing popularity of immortelle (Helichrysum italicum (Roth) G. Don) and its products, particularly in the cosmetic industry, is evident nowadays. This plant is a source of coumarins, especially scopoletin, which are highly soluble in supercritical CO2. Objective The objective of this study was to perform the supercritical CO2 extraction process of Helichrysum italicum flowers at different values of pressure and temperature and to optimise the extraction process using response surface methodology in terms of obtaining the highest extraction yield and yield of extracted scopoletin. Methodology Extraction was performed in a supercritical extraction system under different extraction conditions of pressure and temperature determined by central composite rotatable design. The mass of flowers in the extractor of 40 g, extraction time of 90 min and CO2 mass flow rate of 1.94 kg/h were kept constant during experiments. Antioxidant activity was determined using the DPPH (1,1-diphenyl-2-picrylhydrazyl) free radical scavenging assay method. Scopoletin concentration was determined by HPLC. Results Changes in extraction conditions affect the extracting results remarkably. The greatest extraction yield (6.31%) and the highest yield of scopoletin (1.933 mg/100 g) were obtained under extraction conditions of 20 MPa and 40°C. Extracts have also proven to possess antioxidant activity (44.0–58.1% DPPH scavenging activity) influenced by both temperature and pressure applied within the investigated parameters. Conclusion The extraction conditions, especially pressure, exhibited significant influence on the extraction yield as well as the yield of extracted scopoletin and antioxidant activity of extracts. Copyright © 2016 John Wiley & Sons, Ltd. Supercritical CO2 extraction of H. italicum flowers at different values of pressure and temperature was performed in order to optimize the extraction process using response surface methodology in terms of getting higher extraction yield and yield of scopoletin. The greatest extraction yield and the highest yield of scopoletin were obtained under extraction conditions of 20 MPa and 40°C. Extracts have also proven to possess antioxidant activity influenced by both temperature and pressure applied within the investigated parameters.
Datum: 20.07.2016


Ultra-high Performance Liquid Chromatography with Photodiode Array and Chemiluminescence Detection for the Determination of Polyphenolic Antioxidants in Erigeron acris L. Extracts

Introduction The quality of herbs is directly related to the presence of polyphenolic antioxidants. This is the first report on the quantification of individual polyphenolic constituents of Erigeron acris L. Objective To develop a new method using ultra-high performance liquid chromatography with photodiode array and chemiluminescence (UHPLC-PDA-CL) detection for the separation and determination of polyphenols in Erigeron acris extracts. Methodology The methanolic extracts from leaves and inflorescences of Erigeron acris were prepared by ultrasound assisted extraction. The chromatographic separation was performed on C18 column packed with 1.7-μm particles. The post-column CL detection was based on the enhancing effect of polyphenols on the CL generated in manganese(IV)–hexametaphosphate–formaldehyde system. Results The UHPLC method allowed to separate polyphenols in a short running time (13 min), which was three times shorter compared with traditional HPLC. The CL detection was characterised by 6–48 times higher sensitivity and up to three times lower detection limits compared to PDA detection. Qualitative and quantitative differences were observed in polyphenolic composition of Erigeron acris extracts. The main components of leaves were scutellarin and chlorogenic acid, whereas in inflorescences quercetin 3-O-glucoside was predominant. Conclusion Coupling of UHPLC with CL detection has been developed for the first time. This advanced chromatographic technique coupled with sensitive CL detection is a powerful approach for the investigation of polyphenolic profiles in natural products. The shorter analysis time and diminished waste generation makes the UHPLC method more environmentally friendly and more cost-effective in comparison with conventional HPLC. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


Comparison of Analytical Methods in Chemometric Fingerprinting of Metallicolous and Non-metallicolous Populations of Echium vulgare L.

Introduction Adverse environmental conditions usually change plant biochemical pathways resulting in accumulation or decreased content of both primary and secondary metabolites. The chemometric fingerprinting analysis proves to be a useful tool to reveal phytochemical differentiation between plants inhabiting heavy metal-contaminated and uncontaminated areas. Objective Development and assessment of four analytical techniques – high performance capillary electrophoresis (HPCE), thin-layer chromatography (TLC), mass spectrometry (MS), and Fourier transform infrared (FTIR) spectroscopy in chemometric fingerprinting of metallicolous and non-metallicolous populations of Echium vulgare L. Material and Methods Twenty-one crude methanol extracts of shoot samples representing three populations of Echium vulgare L., two originating from highly metal polluted areas and one from an unpolluted area, were investigated using four analytical methods: HPCE, TLC, MS, and FTIR spectroscopy. Data pre-processing (denoising, background subtracting, horizontal alignment) followed by principal component analysis (PCA), hierarchical clustering analysis (HCA), and phytochemical difference index (DI) calculations facilitated exploration of the differences and similarities between the populations. Results Clear phytochemical divergence between metallicolous and non-metallicolous populations of Echium vulgare was found. The suitability of the analytical techniques for revealing phytochemical markers and discrimination of individuals originating from different populations differed and in general increased in the order: TLC < MS = HPCE < FTIR. Conclusion The chemometric methods applied were successful in discrimination between samples from polluted and unpolluted areas, showing a potential perspective for environmental quality control. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


Profiling and Simultaneous Quantitative Determination of Anthocyanins in Wild Myrtus communis L. Berries from Different Geographical Areas in Sardinia and their Comparative Evaluation

Introduction Myrtus communis L. (Myrtaceae) is a self-seeded shrub, widespread in Sardinia, with anti-inflammatory, antiseptic, antimicrobial, hypoglycemic and balsamic properties. Its berries, employed for the production of sweet myrtle liqueur, are characterised by a high content of bioactive polyphenols, mainly anthocyanins. Anthocyanin composition is quite specific for vegetables/fruits and can be used as a fingerprint to determine the authenticity, geographical origin and quality of raw materials, products and extracts. Objective To rapidly analyse and determine anthocyanins in 17 samples of Myrtus communis berries by developing a platform based on the integration of UHPLC–MS/MS quantitative data and multivariate analysis with the aim of extracting the most information possible from the data. Methodology UHPLC-ESI-MS/MS methods, working in positive ion mode, were performed for the detection and determination of target compounds in multiple reaction monitoring (MRM) mode. Optimal chromatographic conditions were achieved using an XSelect HSS T3 column and a gradient elution with 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Principal component analysis (PCA) was applied to the quantitative data to correlate and discriminate 17 geographical collections of Myrtus communis. Results The developed quantitative method was reliable, sensitive and specific and was successfully applied to the quantification of 17 anthocyanins. Peonidin-3-O-glucoside was the most abundant compound in all the extracts investigated. Conclusion The developed methodology allows the identification of quali-quantitative differences among M. communis samples and thus defines the quality and value of this raw material for marketed products. Moreover, the reported data have an immediate commercial value due to the current interest in developing antioxidant nutraceuticals from Mediterranean plants, including Sardinian Myrtus communis. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 20.07.2016


Pro-toxic 1,2-Dehydropyrrolizidine Alkaloid Esters, Including Unprecedented 10-Membered Macrocyclic Diesters, in the Medicinally-used Alafia cf. caudata and Amphineurion marginatum (Apocynaceae: Apocynoideae: Nerieae and Apocyneae)

Introduction Within the Apocynoideae (Apocynaceae) pro-toxic dehydropyrrolizidine alkaloids have been reported only in Echiteae. However, attraction of pyrrolizidine alkaloid-pharmacophagous insects suggested their presence in Alafia cf. caudata Stapf (Nerieae: Alafiinae) and Amphineurion marginatum (Roxb.) D.J. Middleton (Apocyneae: Amphineuriinae), both used as medicinal plants. Objective To confirm the presence of dehydropyrrolizidine alkaloids in Alafia cf. caudata and Amphineurion marginatum and identify their structures. Methods Methanol extracts of air-dried roots, stems and leaves of non-flowering plants were analysed using HPLC-ESI(+)MS and MS/MS or collision-induced dissociation MS in low and/or high resolution modes. Pyrrolizidine alkaloids were tentatively identified based on the mass spectrometry data. Solid phase extraction combined with semi-preparative HPLC were used to isolate major alkaloids. Structures were elucidated using NMR spectroscopy. Results Monoesters of retronecine with senecioic, hydroxysenecioic or syringic acids were identified in roots of Alafia cf. caudata. Two unprecedented 10-membered macrocyclic dehydropyrrolizidine alkaloid diesters were isolated from roots of Amphineurion marginatum. Pyrrolizidine alkaloids were detected in root and leaf material of Alafia cf. caudata at 0.34 and 0.01% dry weight (DW), and 0.13, 0.02 and 0.09% DW in root, leaf and stem material of Amphineurion marginatum. Conclusions The presence of pro-toxic dehydropyrrolizidine alkaloids suggests that medical preparations of these plants pose potential health risks to consumers. Dehydropyrrolizidine alkaloids are evidently more widespread in Apocynoideae than previously assumed, and it would seem rewarding to study other members of this family for the presence of pyrrolizidines, dehydropyrrolizidines and dihydropyrrolizines. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 19.07.2016






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