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

Rapid Detection and Characterisation of Triterpene Saponins from the Root of Pulsatilla chinensis (Bunge) Regel by HPLC-ESI-QTOF-MS/MS

Introduction Triterpene saponins are the major bioactive components in the root of Pulsatilla chinensis (Bunge) Regel (RPC), and have been reported to possess antitumor and immunological adjuvant activities. However, the isolation, purification and elucidation procedures of triterpene saponins from RPC are difficult and time consuming due to high polarity and structural similarity. Objectives To develop an analytical strategy for discovering and elucidating triterpene saponins in RPC. Methods Methanolic extract of RPC is analysed by high-performance liquid chromatography coupled to electrospray ionisation and quadrupole time-of-flight-mass spectrometry (HPLC-ESI-QTOF-MS/MS). The MS and MS/MS experiments are conducted using the negative-ionisation mode, in order to provide molecular-mass information and production spectra for the structural elucidation of compounds. Results Based on retention times, accurate mass and mass spectrometric fragmentation, 24 triterpene saponins are identified or tentatively elucidated from RPC, of which nine triterpene saponins were not reported previously. Conclusion The HPLC-ESI-QTOF-MS/MS could be employed as a rapid, effective technique to screen and identify triterpene saponins in RPC without tedious and time-consuming isolation of pure constituents. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Issue Information

No abstract is available for this article.
Datum: 17.06.2016

Application of a Smartphone Metabolomics Platform to the Authentication of Schisandra sinensis

Introduction Herbal medicines have been used for a long time all around the world. Since the quality of herbal preparations depends on the source of herbal materials, there has been a strong need to develop methods to correctly identify the origin of materials. Objective To develop a smartphone metabolomics platform as a simpler and low-cost alternative for the identification of herbal material source. Methodology Schisandra sinensis extracts from Korea and China were prepared. The visible spectra of all samples were measured by a smartphone spectrometer platform. This platform included all the necessary measures built-in for the metabolomics research: data acquisition, processing, chemometric analysis and visualisation of the results. The result of the smartphone metabolomics platform was compared to that of NMR-based metabolomics, suggesting the feasibility of smartphone platform in metabolomics research. Results The smartphone metabolomics platform gave similar results to the NMR method, showing good separation between Korean and Chinese materials and correct predictability for all test samples. Conclusion With its accuracy and advantages of affordability, user-friendliness, and portability, the smartphone metabolomics platform could be applied to the authentication of other medicinal plants. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Facile Separation of 5-O-Galloylquinic Acid from Chinese Green Tea Extract using Mesoporous Zirconium Phosphate

Introduction 5-O-Galloylquinic acid from green tea and other plants is attracting increasing attention for its antioxidant and antileishmanial bioactivities. It is always isolated using a silica column, a Sephadex column and high-performance liquid chromatography (HPLC) methods, which are either laborious or instrument dependent. Objective To develop a new method to easily separate 5-O-galloylquinic acid. Methodology Mesoporous zirconium phosphate (m-ZrP) was prepared to conveniently separate 5-O-galloylquinic acid from Chinese green tea extract, and the target compound was easily obtained by simple steps of adsorption, washing and desorption. The effects of the green tea extraction conditions, extract concentrations, and m-ZrP adsorption/desorption dynamics on the 5-O-galloylquinic acid separation were evaluated. Results 5-O-Galloylquinic acid that was separated from a 70% ethanol extract of green tea was of moderate HPLC purity (92%) and recovery (88%), and an increased non-specific binding of epigallocatechin gallate (EGCG) on m-ZrP was observed in the diluted tea extract. The times for maximal adsorption of 5-O-galloylquinic acid in 70% ethanol extract and maximal desorption of 5-O-galloylquinic acid in 0.4% phosphoric acid solution were confirmed as 7 h and 5 h, respectively. Conclusion A facile method to separate 5-O-galloylquinic acid from Chinese green tea extract using m-ZrP was established. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Antitrypanosomal Activity of a Novel Taccalonolide from the Tubers of Tacca leontopetaloides

Introduction Several taccalonolides with various bioactivities have been isolated from Tacca species but no studies to isolate taccalonolides with anti-trypanosomal activity from Tacca leontopetaloides have been reported. Objectives To analyse extracts of the roots of Tacca leontopetaloides, purify the extracts by column chromatography and identify isolated compounds by spectroscopic methods. The compounds and fractions will be tested for antitrypanosomal activity in vitro against Trypanosoma brucei brucei. Material and methods Dried roots or tubers of Tacca leontopetaloides, chromatographic separation and spectroscopic identification. Results A novel taccalonolide A propanoate and some known taccalonolides were isolated and their structures were determined by NMR and mass spectrometry Conclusion Several taccalonolides were isolated from Tacca leontopetaloides and were found to have in vitro antitrypanosomal activity against Trypanosoma brucei brucei and EC50 values for the isolated compounds were from 0.79 µg/mL. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Issue Information

No abstract is available for this article.
Datum: 17.06.2016

In situ Identification of Labile Precursor Compounds and their Short-lived Intermediates in Plants using in vivo Nanospray High-resolution Mass Spectrometry

Introduction Many secondary metabolites in plants are labile compounds which under environmental stress, are difficult to detect and track due to the lack of rapid in situ identification techniques, making plant metabolomics research difficult. Therefore, developing a reliable analytical method for rapid in situ identification of labile compounds and their short-lived intermediates in plants is of great importance. Objective To develop under atmospheric pressure, a rapid in situ method for effective identification of labile compounds and their short-lived intermediates in fresh plants. Methodology An in vivo nanospray high-resolution mass spectrometry (HR-MS) method was used for rapid capture of labile compounds and their short-lived intermediates in plants. A quartz capillary was partially inserted into fresh plant tissues, and the liquid flowed out through the capillary tube owing to the capillary effect. A high direct current (d.c.) voltage was applied to the plant to generate a spray of charged droplets from the tip of the capillary carrying bioactive molecules toward the inlet of mass spectrometer for full-scan and MS/MS analysis. Results Many labile compounds and short-lived intermediates were identified via this method: including glucosinolates and their short-lived intermediates (existing for only 10 s) in Raphanus sativus roots, alliin and its conversion intermediate (existing for 20 s) in Allium sativum and labile precursor compound chlorogenic acid in Malus pumila Mill. Conclusion The method is an effective approach for in situ identification of internal labile compounds and their short-lived intermediates in fresh plants and it can be used as an auxiliary tool to explore the degradation mechanisms of new labile plant compounds. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Identification of Alkaloids in Stephania hainanensis by Liquid Chromatography Coupled with Quadrupole Time-of-flight Mass Spectrometry

Introduction Plants in the genus Stephania can produce diverse bioactive alkaloids. Stephania hainanensis is a medicinal plant that contains effective alkaloids. However, only 10 alkaloids have been reported in this species. Objective To characterise the alkaloids in Stephania hainanensis using liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (LC-QTOF-MS/MS). Methods An LC-QTOF-MS/MS method was developed for structural characterisation of the alkaloids in Stephania hainanensis. The chromatographic separation was performed on a phenyl column with gradient elution, and the tandem mass spectra were obtained by using an electrospray ionisation (ESI) interface in positive ionisation mode. Compound identification was based on the exact masses, fragmentation pathways, retention behaviours and related botanical biogenesis. Results A total of 37 tetrahydroprotoberberine-, quaternary protoberberine-, aporphine-, proaporphine-, benzylisoquinoline- or bisbenzylisoquinoline-type alkaloids were identified or tentatively identified in a single LC run. Twenty-seven of these alkaloids, including the benzylisoquinoline-type of alkaloids, have not been previously reported in Stephania hainanensis. The possible fragmentation pathways of different types of alkaloids were proposed. Besides the general fragmentations, the characteristic losses of CH3N = CH2 were observed for the benzylisoquinoline and aporphine alkaloids with two methyl groups on the nitrogen. Conclusion The LC-QTOF-MS/MS method enabled profiling and rational, but tentative, identification of diverse alkaloids in Stephania hainanensis. The results obtained may be helpful for understanding the bioactivity of S. hainanensis and evaluating the quality of this plant. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Direct Coupling of HPTLC with MALDI-TOF MS for Qualitative Detection of Flavonoids on Phytochemical Fingerprints

Introduction Thin layer chromatographic fingerprints of plant raw materials and extracts for food and pharma applications often focus on phenol carbonic acids and flavonoids. The visual detection and comparison of Rf values of applied reference substances only renders limited phytochemical information. Recently, direct coupling of TLC with MALDI-TOF MS has been successfully applied for analysis of biologically relevant compounds such as lipids. The mass analysis of low molecular weight TLC or HPTLC fingerprints of flavonoids has, to our knowledge, not yet been investigated. Objectives In this study, the feasibility of direct coupling of HPTLC with UV-MALDI-TOF MS for determination of molecular mass of the ubiquitously present flavonol glycoside, rutin, and flavone glycoside, luteolin-7-O-glucoside, as well as their corresponding aglycones, quercetin and luteolin, is demonstrated. Methodology HPTLC plate suitable for combination with a MALDI MS adapter was used for chromatographic separation of compounds of interest. After separation, the plate was sprayed with 2,5 dihydroxybenzoic acid as a MALDI matrix using an automated spraying device. After drying, the developed chromatograms were scanned by UV-MALDI-TOF MS in positive mode with a spatial resolution of 0.2 mm. Results All compounds studied were distinctly detected in MALDI-TOF mass spectra. This is particularly pertinent for the co-eluted aglycones luteolin and quercetin, which could not have been distinguished by the common visual HPTLC derivatisation and evaluation. Conclusion This study demonstrates the potential of MALDI-TOF MS for the analysis of low molecular weight fingerprints of flavonoids directly from their HPTLC chromatogram. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Simultaneous Quantification of Seven Bioactive Flavonoids in Citri Reticulatae Pericarpium by Ultra-Fast Liquid Chromatography Coupled with Tandem Mass Spectrometry

Introduction Citri Reticulatae Pericarpium (CRP) is a commonly-used traditional Chinese medicine with flavonoids as the major bioactive components. Nevertheless, the contents of the flavonoids in CRP of different sources may significantly vary affecting their therapeutic effects. Thus, the setting up of a reliable and comprehensive quality assessment method for flavonoids in CRP is necessary. Objective To set up a rapid and sensitive ultra-fast liquid chromatography coupled with tandem mass spectrometry (UFLC-MS/MS) method for simultaneous quantification of seven bioactive flavonoids in CRP. Methods A UFLC-MS/MS method coupled to ultrasound-assisted extraction was developed for simultaneous separation and quantification of seven flavonoids including hesperidin, neohesperidin, naringin, narirutin, tangeretin, nobiletin and sinensetin in 16 batches of CRP samples from different sources in China. Results The established method showed good linearity for all analytes with correlation coefficient (R) over 0.9980, together with satisfactory accuracy, precision and reproducibility. Furthermore, the recoveries at the three spiked levels were higher than 89.71% with relative standard deviations (RSDs) lower than 5.19%. The results indicated that the contents of seven bioactive flavonoids in CRP varied significantly among different sources. Among the samples under study, hesperidin showed the highest contents in 16 samples ranged from 27.50 to 86.30 mg/g, the contents of hesperidin in CRP-15 and CRP-9 were 27.50 and 86.30 mg/g, respectively, while, the amount of narirutin was too low to be measured in some samples. Conclusion This study revealed that the developed UFLC-MS/MS method was simple, sensitive and reliable for simultaneous quantification of multi-components in CRP with potential perspective for quality control of complex matrices. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Myrosinase Compatible Simultaneous Determination of Glucosinolates and Allyl Isothiocyanate by Capillary Electrophoresis Micellar Electrokinetic Chromatography (CE-MEKC)

Introduction The functional food Cruciferous vegetables contain glucosinolates which are decomposed by the myrosinase enzyme upon tissue damage. The isothiocyanates are the most frequent decomposition products. Because of their various bioactivities, these compounds and the myrosinase is of high interest to many scientific fields. Objective Development of a capillary electrophoresis method capable of myrosinase-compatible, simultaneous quantification of glucosinolates and isothiocyanates. Methods Capillary electrochromatography parameters were optimised, followed by optimisation of a myrosinase-compatible derivatisation procedure for isothiocyanates. Vegetable extracts (Brussels sprouts, horseradish, radish and watercress) were tested for myrosinase activity, glucosinolate content and isothiocyanate conversion rate. Allyl isothiocyanate was quantified in some food products. Results The method allows quantification of sinigrin, gluonasturtiin and allyl isothiocyanate after myrosinase compatible derivatisation in-vial by mercaptoacetic acid. The chromatograhpic separation takes 2.5 min (short-end injection) or 15 min (long-end injection). For the tested vegetables, measured myrosinase activity was between 0.960–27.694 and 0.461–26.322 µmol/min/mg protein, glucosinolate content was between 0–2291.8 and 0–248.5 µg/g fresh weight for sinigrin and gluconastrutiin, respectively. The possible specificity of plants to different glucosinolates was also shown. Allyl isothiocyanate release rate was different in different vegetables (73.13 − 102.13%). The method could also be used for quantification of allyl isothiocyanate from food products. Conclusions The presented capillary electrophoresis method requires a minimal amount of sample and contains only a few sample preparation steps, and can be used in several applications (glucosinolate determination, myrosinase activity measurement, isothiocyanate release estimation). Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 17.06.2016

Chemotaxonomic Studies of Nine Gentianaceae Species from Western China Based on Liquid Chromatography Tandem Mass Spectrometry and Fourier Transform Infrared Spectroscopy

Introduction Gentianaceae species which widely occur all over the world are used as folk medicine and raw food material with bitter properties. Although comparative analysis on metabolites in several Gentianaceae species has been reported, metabolic similarities used for chemotaxonomic studies are not yet clear. Objective To systematically characterise the variations of holistic metabolome and characteristic metabolites (iridoid glycosides and phenols) in nine Gentianaceae species from western China. Methodology Fourier transform infrared (FT-IR) spectroscopy was applied to determine the variations of holistic metabolome. A targeted metabolic profiling using liquid chromatography with tandem mass spectrometry (LC–MS/MS) was established for determination of seven characteristic metabolites and identification of their derivatives. Both FT-IR and LC–MS/MS data were subjected to chemometrics analysis for exploring variations in iridoid glycosides and phenols within these species. Results Holistic metabolome in genera Gentiana and Swertia was largely different. Diversity of the biosynthetic pathway of iridoid glycosides was also observed in these species. Principal component analysis (PCA) showed a clear separation according to infrageneric classifications of genus Gentiana. Some secondary metabolites, such as mangiferin, rhodenthoside A-C, isoorientin, isovitexin, amarogentin, and swertianolin would serve as potential chemotaxonomic markers to differentiate Gentianaceae species. Furthermore, the accumulation of the six major metabolites seems to depend on geographical regions in Sect. Monopodiae and Sect. Cruciata. Conclusions The combination of LC–MS/MS and FT-IR would provide some potential evidence on chemotaxonomic studies of Gentianaceae. Copyright © 2016 John Wiley & Sons, Ltd.
Datum: 26.02.2016

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