Identification Of Flavonoids Research Articles

TLC-MALDI-TOF-MS-based identification of flavonoid compounds using an inorganic matrix.

Thin layer chromatography (TLC) is frequently used to obtain the fingerprint of a plant extract. Although the retardation factor and the response to visualisation give primary information about compound identification, the direct TLC-mass spectrometry (MS) coupling allows a more detailed characterisation of samples. To demonstrate the potential for the flavonoid dereplication using an inorganic matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) method with and without TLC separation. Samples derived from wine, apple or rose were deposited on an aluminium-backed silica gel TLC sheet compatible with the MS adapter. Unlike the wine sample, for apple and rose samples compound derivatisation was necessary. These two samples were deposited twice and the plate was cut in two parts. One half was oversprayed with Neu-Peg reagent to visualise flavonoids while the inorganic matrix was deposited on each flavonoid zone on the second half for MS ionisation. Mass spectra obtained for samples without plate development showed numerous ions corresponding to glycosylated flavonoids. The lower m/z observed could be due either to aglycone flavonoids or to in-source fragment ions. After plate development, a separation of many spots was observed and each spot was analysed separately leading to a deeper identification of the present flavonoids. Moreover, isobaric flavonoids with different hRf values could be differentiated. TLC-MALDI-TOF-MS using an inorganic matrix enabled the analysis of anthocyanins in positive mode and of flavonols, flavanols, dihydrochalcones and phenolic acids in negative mode, reducing adduct, aggregate forms giving thus simple and reliable spectra for the dereplication approach of flavonoids in complex samples.

Identification of flavonoids in Plumula nelumbinis and evaluation of their antioxidant properties from different habitats

Plumula nelumbinis is a well-known functional food and traditional medicine. In this study, a sensitive and efficient ultra-performance liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOF-MS) method was used for systematic identification of the total flavonoids in Plumula nelumbinis. Based on comparisons of retention times, mass spectrometry data and molecular weights with standards and references, thirty-eight flavonoid compounds were unambiguously or tentatively identified. Ten flavonoids were detected for the first time in Nelumbo and fifteen flavonoids were detected for the first time in Plumula nelumbinis. The study revealed a positive correlation between total flavonoids and the results of the antioxidant activity determinations which were accomplished with 1,1-diphenyl-2-picrylhydrazyl (DPPH), oxygen radical absorbance capacity (ORAC) and ferric reducing ability of plasma (FRAP) assays. In 13 Chinese habitats that were examined for the first time, the content of flavonoids varied from 44.51 ± 0.58 to 117.82 ± 0.35 μg RE/mg DW. Among of them, Hunanxiangtan and Shandongheze provinces produce plants with the highest content of flavonoids, and also exhibiting the strongest antioxidant activity measured in the three assays. These two habitats could therefore have advantages over other habitats. This study applies rational and effective utilities to the analysis of Plumula nelumbinis samples from different habitats and allows their evaluation as natural medicines or functional food materials in commercial processing.

Isolation and Identification of Flavonoids in Iraqi <i>Rheum ribes</i> Root

Isolation and Identification of Flavonoids in Iraqi <i>Rheum ribes</i> Root

An optimized solvent extraction and characterization of unidentified flavonoid glucuronide derivatives from spinach by UHPLC-HR-QTOF-MS

A rapid, sensitive analytical method using ultra-high-pressure liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-HR-QTOF-MS) was developed for the identification and quantification of flavonoids from spinach. The extraction efficiency of flavonoids was evaluated by different solvents such as acetone, ethanol, methanol, acetone: water (70:30), ethanol: water (70:30) and methanol: water (70:30). Flavonoid identification was achieved by UV spectra, high resolution accurate mass and their fragmentation pattern. The precursor and product ions were recorded by both broadband collision ion dissociation (bbCID) and multiple reaction monitoring (MRM) techniques. Different collision energies (5, 10, 15, 20, 40, and 70 eV) were optimized to obtain the mass spectra of flavonoids in positive and negative ionization modes. For the first time, five minor flavonoid glucuronide derivatives were identified in spinach. MRM and bbCID provided glucuronide fingerprint ions at m/z 175.0278 and m/z 113.0257 respectively in negative ionization mode. The quantification of identified flavonoids was achieved by 5,3',4'-trihydroxy-3-methoxy-6:7-methylen-dioxyflavone-4'-β-D-glucuronide which was purified by semi-preparatory HPLC. The purity of the isolated compound was confirmed by NMR analysis. The identified 5,3',4'-trihydroxy-3-methoxy-6:7-methylen-dioxyflavone-4'-β-D-(2′-O-feurloyl-glucuronide) was the prominent flavonoid and the level was significantly higher in the acetone fraction (2.95 ± 0.16 µg/g FW). This study demonstrates the systematic identification of potential bioactive compounds especially glucuronide derivatives from spinach.

Evaluation of the effect of arbuscular mycorrhizal fungi infection in three aromatic plants

The aim of this study was to evaluate the effect of the colonization of arbuscular mycorrhizal fungi in the production of secondary metabolites in Petroselinum crispum, Salvia officinalis and Dysphania ambrosioides. Three treatments were established: Control (C), commercial inoculum (CI) and native inoculum (NI). Every 15 days height, stem diameter and number of leaves were measured. The percentage of colonization was assessed by staining of roots, and identification of flavonoids through TLC thin layer chromatography, finally the concentration of total phenols was evaluated by spectrophotometry from the ethanol extracts of each plant. The results of development of biomass and the percentage of colonization show statistically significant difference for the three variables evaluated (P≤0.05) from each culture in the treatment of NI with respect to control, showing a greater effect on plants of S. officinalis (51% and 91%, respectively). Chromatography reveals the presence of flavonoids in the three plants; however this is more intense for the treatment of NI, showing increased production of total phenols in S. officinalis. Finally, the chemical characterization of the substrate shows a higher assimilation of nitrogen and phosphorus (0.11% and 0.35 mg / kg respectively) in S. officinalis associated with a native inoculum (NI).

UFLC-Q-TOF-MS/MS-Based Screening and Identification of Flavonoids and Derived Metabolites in Human Urine after Oral Administration of Exocarpium Citri Grandis Extract.

Exocarpium Citri grandis (ECG) is an important Traditional Chinese Medicine (TCM) for the treatment of cough and phlegm, and the flavonoids contained were considered the main effective components. To date, the systematic chemical profiling of these flavonoids and derived in vivo metabolites in human have not been well investigated. ECG was extracted using boiling water and then provided to volunteers for oral administration. Following the ingestion, urine samples were collected from volunteers over 48 h. The extract and urine samples were analyzed using ultra-fast liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry (UFLC-Q-TOF-MS/MS) system to screen and identify flavonoids and derived in vivo metabolites. A total of 18 flavonoids were identified in the ECG extract, and 20 metabolites, mainly glucuronide and sulfate conjugates, were screened in urine samples collected post consumption. The overall excretion of naringenin metabolites corresponded to 5.45% of intake and occurred mainly within 4–12 h after the ingestion. Meanwhile, another 29 phenolic catabolites were detected in urine. Obtained data revealed that flavonoids were abundant in the ECG extract, and these components underwent extensive phase II metabolism in humans. These results provided valuable information for further study of the pharmacology and mechanism of action of ECG.

Shading and vermicompost effect on growth and flavonoid content of Tapak Liman (Elephantopus scaber L.)

Tapak Liman (Elephantopus scaber L) is one of Indonesian medicinal plants which is well known as weed. In Thailand, Tapak Limanthis plant is use for traditional medicine due to its flavonoids contains. Flavonoid is compound with red, yellow, purple and blue pigments, used for cancer, aphrodiasiac and anti-radical treatments. One obstacle of Tapak liman cultivation is the effort to increase its flavonoids compound. There is a bridge between flavonoids compound with growth and yield of Tapak Liman. For that, this research aims to find out the effect of shade intensity combined with vermicompost dosage on Tapak Liman growth and yield. This research was conducted in Mei to August 2016 at Medicinal Plantation of BPTO, Tanjungsari Village, Tegal Gede, Karanganyar. Complete Randomized Design compiled with split plot and two factors: shade intensity (0%, 50%, 75%) and vermicompost dosage per plant (0 g, 250 g, 500 g, 750 g) used as the experimental design. The variables observed are leaves number,leaves length, canopy diameter, fresh weight, dry weight, root length, chlorophyl analysis and flavonoid identification. Data were analyzed using ANOVA, any significant treatments followed with Duncan’s Multiple Range Test (DMRT) at α = 10%. Result showed that 75% shade intesity and 750 g of vermicompost has gave highest yield of leaf and total simplicia of Tapak Liman. Shade intensity of 50% with 250 g of and 500 g/plant of vermicompost dosage showed highest flavonoid rendement (Rf 0,5) with highly contrasting spot colors.

Extraction, isolation and identification of flavonoid from Chenopodium album aerial parts

Chenopodium album L., (C. album) (family: Chenopodiaceae) is an annual shrub widely grown in Asia, Africa, Europe and North America. It is commonly known as Bathua (in Hindi), pigweed, fat hen or lamb-quarters. The leaves of C. album are applied as a poultice to bug bites, sunstroke, rheumatic joints and as mild laxative. The flavonoids contained in C. album aerial parts were extracted, identified and characterized. Sequential soxhlet extraction was subjected to preliminary phytochemical screening and flavonoid quantification. The results showed that maximum yield of the flavonoid (7.335 mg/g) were obtained from acetone extract. This acetone extract was subjected to flash chromatography for isolation of flavonoid. Characterization of isolated flavonoid was done by UV, IR, 1H & 13C NMR and MS. On the basis of chemical and spectral analysis structure was elucidated as 2-(3, 4-dihydroxyphenyl)-3, 5, 7-trihydroxy-4H-chromen-4-one, a flavonoid.

Phytochemical screening and antioxidant activity of ethanolic extract and ethyl acetate fraction from basil leaf (Ocimum basilicum L.) by DPPH radical scavenging method

Basil leaf (Ocimum basilicum L.) contains various compounds such as flavonoid, alkaloid, phenol and essential oil, so it needs to be fractionated to find out the flavonoid compound with the greatest potential as an antioxidant. This research was aimed to know the chemical compound, antioxidant potential of ethanolic extract and ethyl acetate fraction from basil leaf. The basil leaf was extracted by maceration using ethanol 70 %. The crude extract was fractionated with ethyl acetate. The ethanolic extract and ethyl acetate fraction were screened of phytochemical content including identification of flavonoids, alkaloids and polyphenolics. The antioxidant activity of the ethanolic extract and ethyl acetate fraction were tested qualitatively with 2,2-diphenyl-1-picrylhydrazyl (DPPH) and phosphomolybdate. Its antioxidant activity was determined quantitatively using DPPH radical scavenging method. Phytochemical screening test showed that ethanolic extract and ethyl acetate fraction from basil leaf contain flavonoids, polyphenolics, and alkaloids. The qualitative analysis of antioxidant activity of ethanolic extract and ethyl acetate fraction from basil leaf showed an antioxidant activity. The IC50 value of ethanolic extract, ethyl acetate fraction and quercetin were 1,374.00±6.20; 389.00±1.00; 2.10±0.01μg/mL, respectively. The research showed that antioxidant activity of the ethyl acetate fraction more potential than the ethanol extract of the basil leaf, but less than quercetin.

Rapid ultra-high-performance liquid chromatography/quadrupole time-of-flight tandem mass spectrometry and selected reaction monitoring strategy for the identification and quantification of minor spinacetin derivatives in spinach.

Spinach is green leafy vegetable which is a rich source of flavonoids, phenolic acids, carotenoids, and vitamins A, C and E. It contains unique flavonoids which have significant anticarcinogenic, antiinflammatory and free radical scavenging activities. The present study reports the systematic identification and quantification of novel flavonoids by ultra-high-performance liquid chromatography with quadrupole time-of-flight tandem mass spectrometry (UHPLC/HR-QTOFMS). An ultrasonication technique was used for the extraction of flavonoids from spinach. A rapid and reliable analytical method was established for the identification of flavonoids from methanolic extract. Flavonoids were characterized by their ultraviolet (UV) spectra, high-resolution accurate masses and MS/MS fragmentation pathways obtained using electrospray ionization (ESI). Furthermore, precursor ions from the intact molecule, and the resulting product ions, were monitored by selected reaction monitoring (SRM) with different collision energies in positive and negative ion mode. For the first time, five minor spinacetin derivatives were identified under optimized SRM and broadband collision-induced dissociation (+bbCID) conditions. Fragmentation pathways were proposed for spectra obtained in ESI positive ion mode. The use of HR-QTOFMS and SRM allowed us to differentiate between molecules with the same nominal mass. The identified spinacetin derivatives were found to be acylated with ferulic and coumaric acids. UHPLC interfaced with HR-QTOFMS in combination with SRM provides a rapid, reliable and highly sensitive method for the identification of flavonoids, and potentially other bioactive compounds, in a complex matrix.

Identification of flavonoids of Wheatgrass (Triticum aestivum L.) at various stages of growth and evaluation of their Antioxidant Activity

Identification of flavonoids of Wheatgrass (Triticum aestivum L.) at various stages of growth and evaluation of their Antioxidant Activity

Development of a Comprehensive Flavonoid Analysis Computational Tool for Ultrahigh-Performance Liquid Chromatography-Diode Array Detection-High-Resolution Accurate Mass-Mass Spectrometry Data.

Liquid chromatography and mass spectrometry methods, especially ultrahigh-performance liquid chromatography coupled with diode array detection and high-resolution accurate-mass multistage mass spectrometry (UHPLC-DAD-HRAM/MSn), have become the tool-of-the-trade for profiling flavonoids in foods. However, manually processing acquired UHPLC-DAD-HRAM/MSn data for flavonoid analysis is very challenging and highly expertise-dependent due to the complexities of the chemical structures of the flavonoids and the food matrixes. A computational expert data analysis program, FlavonQ-2.0v, has been developed to facilitate this process. The program first uses UV-vis spectra for an initial stepwise classification of flavonoids into classes and then identifies individual flavonoids in each class based on their mass spectra. Step-wise identification of flavonoid classes is based on a UV-vis spectral library compiled from 146 flavonoid reference standards and a novel chemometric model that uses stepwise strategy and projected distance resolution (PDR) method. Further identification of the flavonoids in each class is based on an in-house database that contains 5686 flavonoids analyzed in-house or previously reported in the literature. Quantitation is based on the UV-vis spectra. The stepwise classification strategy to identify classes significantly improved the performance of the program and resulted in more accurate and reliable classification results. The program was validated by analyzing data from a variety of samples, including mixed flavonoid standards, blueberry, mizuna, purple mustard, red cabbage, and red mustard green. Accuracies of identification for all samples were above 88%. FlavonQ-2.0v greatly facilitates the identification and quantitation of flavonoids from UHPLC-HRAM-MSn data. It saves time and resources and allows less experienced people to analyze the data.

Isolation and identification of flavonoids from anticancer and neuroprotective extracts of <i>Trigonella foenum</i> graecum

Purpose: To evaluate the protective effects of Trigonella foenum graecum methanol and ethyl acetate extracts, against cancer cell lines and NaNO 2 -induced neurodegeneration in mice brain. Methods: Adult male albino mice ( n = 20) were administered NaNO 2 orally at a dose of 300 mg/kg for 15 days. The control group received distilled water and normal mice feed. Experimental groups were given T. foenum graecum methanol and ethyl acetate extracts in two different doses of 100 and 200 mg/kg orally for 15 days. Histopathological examination of the brain was carried out with the aid of cresyl violet and H&E staining. In addition, the cytotoxicity of the extracts was evaluated by 3-(4,5- dimethylthiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay against HCEC, MCF-7 and Hep2 cell lines. Nuclear magnetic resonance (NMR) and electrospray ionization mass spectrometry (ESI-MS) were used to determine the structures of the bioactive compounds. Results: Methanol and ethyl acetate extracts of T. foenum graecum seeds inhibited neurodegeneration in the hippocampus and cortex regions of the brain when compared to control group. Moreover, the extracts exhibited anticancer activity against Hep2 and MCF-7cells and low cytotoxicity against HCEC, sparing healthy cells in-vitro . In addition, two flavonoids amurensin and cosmosiin were isolated from T. foenum graecum extracts. Conclusion: Amurensin and cosmosiin from T. foenum extracts are reported here for the first time agents that possess significant anticancer and neuroprotective properties. Keywords: Trigonella foenum-graecum , Anticancer, Neurodegeneration, Flavonoids, Amurensin, Cosmosiin

Phytochemical Screening and Antioxidant Activity of Selekop (Lepisanthes amoena) Fruit

Selekop (Lepisanthes amoena (Hassk.) Leenh.) plant leaves are used by the Dayak tribe of East Kalimantan as traditional cosmetics. Selekop fruit is also edible, but not well known. This study was conducted to obtain the phytochemical content and antioxidant assay in flesh, seed and pericarp extracts from the fruit of Selekop. Phytochemical analysis was conducted on ethanol extract for identification of flavonoid, alkaloid, tannin, saponin, triterpenoid and steroid. The antioxidant activity was done by DPPH assay with ascorbic acid as positive control. The flesh contained flavonoid, saponin, and tannin; the seed contained flavonoid, alkaloid, saponin, triterpenoid, and tannin; and the pericarp contained flavonoid, alkaloid, saponin, triterpenoid, and tannin. Analysis of antioxidant activity revealed the following Inhibitory Concentration (IC50 values): 122.51 ppm of flesh, 63.30 ppm of seed, 53.21 ppm of pericarp and 3.06 ppm of ascorbic acid. Based on these results, the ethanol extract of the seed and the flesh had a phytochemical content and antioxidant activity which was better than the flesh extract from Selekop fuit.

Isolation and identification of flavonoid from mycorrhizal and non mycorrhizal roots and rhizomes of Acorus calamus

Isolation and identification of flavonoid from mycorrhizal and non mycorrhizal roots and rhizomes of Acorus calamus

FlavonoidSearch: A system for comprehensive flavonoid annotation by mass spectrometry

Currently, in mass spectrometry-based metabolomics, limited reference mass spectra are available for flavonoid identification. In the present study, a database of probable mass fragments for 6,867 known flavonoids (FsDatabase) was manually constructed based on new structure- and fragmentation-related rules using new heuristics to overcome flavonoid complexity. We developed the FlavonoidSearch system for flavonoid annotation, which consists of the FsDatabase and a computational tool (FsTool) to automatically search the FsDatabase using the mass spectra of metabolite peaks as queries. This system showed the highest identification accuracy for the flavonoid aglycone when compared to existing tools and revealed accurate discrimination between the flavonoid aglycone and other compounds. Sixteen new flavonoids were found from parsley, and the diversity of the flavonoid aglycone among different fruits and vegetables was investigated.

UPLC fingerprint analysis and identification of flavonoids in Scutellaria indica by UFLC-triple-quadrupole time-of-flight mass spectrometry

ABSTRACTThe aim of this study was to raise the quality control level of Scutellaria indica. A quick and stable ultra performance liquid chromatography method was established for the fingerprint analysis of S. indica. A total of 32 common peaks were marked with 10 batches S. indica detected in 30 min using similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine (2004A version). Besides, a high-resolution mass spectrometer was used to identify flavonoids in S. indica. A total of 27 flavonoids in S. indica were identified. And a series of fragmentation regularities were obtained, which could be used for the identification of other flavonoids. Therefore, the established liquid chromatography–tandem mass spectrometer method could be successfully utilized for the quality control of S. indica.

Construction of Comprehensive Flavonoid Analysis Tool by Using UV‐vis Spectra Library, In‐house Database, and Chemometrics

Liquid Chromatography and mass spectrometry methods, especially ultra‐high performance liquid chromatography coupled with high resolution accurate mass‐mass spectrometry (UHPLC‐HRAM‐MS), have become the best methods for flavonoid identification and quantification. However, processing acquired UHPLC‐HRAM‐MS data for flavonoid analysis is very challenging and highly expertise‐dependent because of the complexity of the physical and chemical properties of the flavonoids. An expert data analysis program, FlavonQ, has been developed to facilitate this process. The program first categorizes the flavonoids using a chemometric model based on the UV‐Vis spectra library compiled for 146 flavonoid reference standards. A novel stepwise classification strategy is used that provides data representation in each step as optimized by a projected distance resolution (PDR) method. The stepwise classification strategy significantly improves the performance of the classifiers which results in more accurate and reliable classification results. An in‐house flavonoid database which contains 5686 previously reported flavonoids is used for identification of flavonoids. FlavonQ was validated by analyzing data from samples with spiked flavonoid mixed standards and plant samples including blueberry, mizuna, purple mustard, red cabbage, and red mustard green extract. Accuracies for identification for all samples were above 88%. FlavonQ greatly facilitates the identification and quantitation of flavonoids from UHPLC‐HRAM‐MS data. The process is automated, saving tremendous resources, and allowing less‐experienced people to perform data analysis on flavonoids with reasonable results.Support or Funding InformationThis research is supported by the Agricultural Research Service of the U.S. Department of Agriculture, an Interagency Agreement with the Office of Dietary Supplements at the National Institutes of Health (Y01 OD001298‐01). The John A. Milner Fellowship program by USDA Beltsville Human Nutrition Research Center and the NIH Office of Dietary Supplements is acknowledged for the support to Dr. Mengliang Zhang. We thank to Dr. Peter de B. Harrington for providing his codes on chemometric models.

Spectral analysis of flavonoids content of dandelion Taraxacum officinale W. and chicory usual Cichorium intybus L. depending on growing places

Article presents results of the identification of flavonoids by chromatographic method, dependence of content flavonoids on conditions of plant growth. Coefficients of technogenic concentrations of heavy metals in soil samples and intensity of migration of pollutants from soil to plants using biological absorption coefficient were established.

Rational development of MS method for rapid identification of Flavonoids in plant extracts

Rational development of MS method for rapid identification of Flavonoids in plant extracts