Identification Of Isoflavones Research Articles

Integrated strategy for identifying isoflavones in Belamcandae Rhizoma based on the combination of mass defect filtering and neutral-loss-triggered multistage fragmentation

Belamcandae Rhizoma is a widely used traditional Chinese herbal medicine with isoflavones as the main active ingredient. In this paper, an integrated strategy was developed to discover and identify new isoflavones in Belamcandae Rhizoma by an ultra-high-performance liquid chromatography coupled with high resolution multistage mass spectrometry. Different characterization methods were used based on structural features of isoflavone aglycones and glycosides. On one hand, we adopted a data-dependent acquisition mode incorporated into intelligent AcquireX deep scan algorithms to analyze crude extract, and used a mass defect filtering technique to filter out two kinds of isoflavone aglycones from the extract. On the other hand, neutral-loss-triggered MSn was used to analyze isoflavone glycosides, and under this acquisition mode, MSn scan only took place when chemical components exhibited specific neutral losses. Identification of isoflavones was achieved either by comparison with reference compounds or analysis of characteristic product ions based on MS2 or MSn fragmentation patterns. UV absorbance spectra also contributed to the confirmation of isoflavones. As a result, a total of 65 isoflavone aglycones (42 new aglycones) and 142 isoflavone glycosides (122 new glycosides) were discovered, including a number of trace components. Meanwhile, modifications of new sugar moieties, such as xylose, rhamnose and 6-O-(4‑hydroxy-3,5-dimethoxybenzoyl)-β-D-glucose, were discovered in Belamcandae Rhizoma for the first time. These results indicated the feasibility of this established strategy for in-depth identification of new isoflavone aglycones and glycosides.

Tailoring enhanced production and identification of isoflavones in the callus cultures of Pueraria thomsonii Benth and its model verification using response surface methodology (RSM): a combined in vitro and statistical optimization

BackgroundScientifically, isoflavones from Pueraria thomsonii Benth possess diverse pharmacological activities and have been used to treat various diseases. In vitro propagation of callus has contributed to the reliability for large-scale production of target compounds. However, the factors affecting the biosynthesis of major isoflavones daidzin, puerarin and daidzein in the callus culture of P. thomsonii are still not known. Therefore, we aimed to enhance the in vitro production of daidzin, puerarin and daidzein by optimizing three independent factors such as temperature, NAA and 6-BA concentrations.ResultsOur findings showed that the optimal concentrations for in vitro biomass production and efficient synthesis of puerarin, daidzin and daidzein were found to be 0.158%, 0.463% and 0.057%, respectively. In addition, the HPLC fingerprint with chemo-metrics analysis was constructed by linear regression of the puerarin, daidzin and daidzein which was found to be in the range of 1.0–36.0, 5.0–72.0 and 1.0–15.0 mg/mL and the LODs and LOQs were found to be 0.15, 0.52, 0.35 and 0.28, 1.50, 0.50 mg/mL for puerarin, daidzin and daidzein, respectively. Surprisingly, our results were also in agreement with the concentration obtained from the model verification for optimal and efficient production of puerarin, daidzin and daidzein which was found to be 0.162%, 0.458% and 0.049%, respectively.ConclusionsIn summary, our present investigation provides new insights that could facilitate the enhanced production of valuable isoflavones in P. thomsonii using plant cell cultures treated with appropriate elicitor combinations and temperature. As far as the authors are concerned, this is the first report on production of daidzin, puerarin and daidzein at higher yield at laboratory level for a wide range of applications in future food, medicinal and pharmaceutical companies.

Identification of isoflavones in the extract of supplements for menopause symptoms by direct infusion electrospray ionization tandem mass spectrometry.

In recent years high consumption of dietary supplements has been observed. However, the consumption of dietary supplements may lead to the unexpected side effects that can be related to the number of adulterated supplements quite often marketed. It has prompted the search for a fast and reliable method of identification of main active compounds in the supplements. In this study, the isoflavones present in the methanol extracts of dietary supplements for menopause symptoms were identified by using direct infusion electrospray ionization tandem mass spectrometry. The product ion spectra of [M-H]- ions derived from the extracts matched very well those obtained for standard compounds. Daidzein, genistein, and glycitein were identified in the soy-based supplements, while daidzein, formononetin, and biochanin A were identified in the red clover-based ones. The respective [M+Na]+ ions were also detected; however, their product ion spectra did not allow isoflavone identification. It can be concluded that the main isoflavones present in the extracts of dietary supplements can be successfully and quickly identified by using the direct infusion electrospray ionization in negative ion mode, followed by the tandem mass spectrometry experiment.

Activity guided isolation of isoflavones and hyphenated HPLC-PDA-ESI-ToF-MS metabolome profiling of Azorella madreporica Clos. from northern Chile

Four isoflavones were isolated from the MeOH extract of a sample of Azorella madreporica from the high Andes of northern Chile by selective fractionation using the bleaching of the DPPH radical. The compounds were identified by NMR spectroscopy and mass spectrometry together with 18 additional complex isoflavones present in the plant extract which were detected for the first time using HPLC hyphenated with UV–visible and high resolution mass detectors (HR-PDA-ToF-ESI-MS). The results obtained showed that the methodology developed is appropriate for rapid analysis and identification of isoflavones in extracts from native A. madreporica which can have potential use as an additive for food, or as a nutritional supplement due to the high content on isoflavones (yield of 162.98, 290.18, 44.25 and 31.03mg/kg dry plant for the isolated compounds genistein-8-C-β-d-glucopyranoside (1), alpinum isoflavone (19), angustone C (21) and 4´-acetyl-alpinum isoflavone, 22, respectively). The latter compound 22 is a new isoflavone derivative isolated for the first time from a plant source. The moderate antioxidant properties and content of isoflavonoids found in the aerial parts can explain, at least in part, the traditional use of the native plant and health benefits of the infusions.

Detecção de soja pelo teor de isoflavonas em lombo injetado de suíno

No presente trabalho, a utilização de 1,5% de proteína isolada de soja em lombo de suíno foi detectada por meio do teor de isoflavonas, utilizando-se cromatografia líquida de alta eficiência. Elegeu-se o pico de genistina para quantificação, construindo-se uma curva-padrão com a proteína isolada de soja utilizada nas salmouras para injeção das carnes. A técnica revelou-se como uma alternativa rápida e eficaz para ser implantada no controle da utilização desse ingrediente em produtos cárneos.

Rapid-resolution HPLC with spectrometric detection for the determination and identification of isoflavones in soy preparations and plant extracts

A rapid-resolution HPLC/UV-VIS DAD separation method (which takes <1 min) for the determination and identification of genistin, genistein, daidzein, daidzin, glycitin, glycitein, ononin, formononetin, sissotrin and biochanin A in fmol quantities in submicroliter sample volumes was optimized. A linear gradient elution (0 min 22% B, 1.0 min 80% B, 1.4 min 100% B, 1.8 min 22% B) using a mobile phase containing 0.2 % (v/v) acetic acid (solvent A) and methanol (solvent B) was applied on a Zorbax SB C18 column (1.8 microm particle size) at 80 degrees C. The method was verified using samples of bits of soy and methanolic extracts from Trifolium pratense, Iresine herbstii and Ononis spinosa plants. Pseudobaptigenin glucoside, irilone, prunetin, texasin, tlatlancuayin and other isoflavones, in addition to aglycones of isoflavones and their beta-glucosides and malonyl and acetyl derivatives, were identified by UV-VIS DAD and electrospray mass spectrometric (ESI-MS) detection in the extracts.

Separation and identification of isoflavones with on‐line liquid chromatography‐electrochemistry‐thermospray mass spectrometry

AbstractThe on‐line high‐performance liquid chromatography‐electrochemistry‐thermospray mass spectrometry (LCEC‐LC/MS) for separation and identification of the phenolic isoflavones genistein, formononetin, and Biochanin A is described. The isoflavones are separated on a C‐18 reversed phase column with methanol: 0.1 M ammonium acetate pH (70:30 v/v) as eluent, which is compatible for both LCEC and LC/MS detection. Low ng detection is achieved with a conventional glassy carbon thin‐layer electrochemical cell at 1.3 V vs. Ag/AgCl. Selected ion monitoring (SIM) mode LCMS results in detection levels at the 20–50 ng range, and the compounds are positively identified. In the on‐line detection, the electrochemical cell is connected to the thermospray and the column effluent flows from the working electrode into the heated thermospray capillary tube. The results show that on‐line coupling between a thin‐layer LCEC and a thermospray‐LC/MS can be used to characterize analytical systems.