Yield Of Total Flavonoids Research Articles

Optimization of Liquid Fermentation Medium for Production of Inonotus sanghuang (Higher Basidiomycetes) Mycelia and Evaluation of their Mycochemical Contents and Antioxidant Activities.

Inonotus sanghuang, an authentic "Sanghuang" mushroom used in traditional Chinese medicine, is known to possess important pharmacological activities. In this study, we aimed to optimize the liquid fermentation medium for I. sanghuang mycelial production and to determine the effects of two-stage cultivation (shake and static) on the yield of total flavonoids, total phenolics, and polysaccharides, as well as the antioxidant activities of I. sanghuang mycelial extracts (ISME). Under an optimized medium composition (38.96 g/L of corn flour, 4.15 g/L of yeast extract, 20.55 g/L of bran and pH 6.39), the predicted and experimental optimal mycelial biomasses were 17.60 g/L and 18.33±0.86 g/L, respectively. The results of two-stage cultivation showed that contents of total flavonoids and total phenolics in mycelia increased by 37.92% and 77.27%, respectively. However, irregular polysaccharide contents were noted throughout the experimental period. Antioxidant assays showed that ISME possessed good free-radical scavenging activity, which is mainly contributed by polyphenolic-type metabolites.

Kinetic Modelling for Flavonoid Recovery from Red Grape (Vitis vinifera) Pomace with Aqueous Lactic Acid

This study was undertaken with the aim of establishing a correlation between the extraction yield in total flavonoids from red grape pomace and the extraction temperature, using 0.5% (w/v) aqueous lactic acid as the solvent system. Extraction of flavonoids was found to obey second-order kinetics, and on such a basis, the yield in total flavonoids at saturation could be very effectively determined and correlated with temperature using non-linear regression. The results indicated that the extraction yield at saturation is highly correlated with temperature, following a quadratic function. The extract obtained at 40 °C had an optimal predicted total flavonoid yield of 13.27 mg rutin equivalents per gram of dry weight, and it was further analyzed by liquid chromatography-mass spectrometry to characterize its major constituents. The polyphenols detected were flavanols, flavonols and an anthocyanin. The outcome of this study outlined that temperatures above 40 °C are rather unfavorable for flavonoid extraction from red grape pomace, as suggested by the model established through kinetics.

Study on Microwave-Assisted Extraction Technology of Total Flavonoid from Castor Leaves

Orthogonal experiment optimization of microwave-assisted extraction (MAE) conditions of total flavonoids from castor leaves with the factors of ethanol concentration, microwave power, extraction time, solid/liquid ratio on the yield of total flavonoids were studied. The results showed that optimum extracting conditions of total flavonoids from castor leaves were as follows: adding 1:50 g·mL-1 of 40% alcohol and extracting for 3 min,microwave irradiation power 464W. Microwave extraction of total flavonoids from castor leaves is fast, energy saving and high extraction rate.

Application of graphene oxide as a catalyst to accelerate extraction of total flavonoids and the hydrolysis of baicalin from Radix scutellaria

A new method, graphene oxide (GO)-assisted ethanol reflux extraction (GERE) was investigated to extract total flavonoids from Radix scutellaria and accelerate the hydrolysis of baicalin (BG) to baicalein (B), simultaneously. The parameters of GERE were optimized by the single factor experiments, the optimal conditions were 1.1mg GO/g dry raw material, ethanol 60% (v/v), ratio of liquid to solid 15 (mL/g) at 70°C for 6h. Under optimized conditions, the yields of total flavonoids and baicalein by GERE reached 24.95% and 0.38%, about 9.72% and 375% higher than single ethanol reflux extraction (ERE), respectively. SEM results revealed that after treated with GO, the Radix scutellaria surface appeared cracks, ravines and small holes. The structural disruption of wall reduces the mass transfer resistance and accelerates the leaching of flavonoids by ethanol solvent. XRD and FTIR results showed that GO reduced the crystallinity and the grain sizes of the Radix scutellaria cell, changed the internal structures of cellulose as well as cellulose accessibility, and improved the hydrolysis efficiency. HPLC results revealed that the content of baicalein in extract by GERE was significantly higher than that by ERE, which means that GO can catalyze the hydrolysis of baicalin to baicalein. The high yields of total flavonoids and baicalein can be related to the catalytic activity of GO, which can lead to the destruction of the cell wall, the reduction of the crystallinity and the grain sizes of cellulose. Thus, it was indicated that GERE method could be an alternative method for natural compounds extraction in industry.

Optimum Process for Extraction of Total Flavonoids from Castor Leaves by Orthogonal Test

Optimization of total flavonoid extraction from Castor leaves using orthogonal test methodology with the factors of ethanol concentration, extraction temperature,extraction time,solid/liquid ratio on the yield of total flavonoids were investigated.The results showed that optimum extracting conditions of total flavonoids from castor leaves were as follows: adding 1:40 g·mL-1 of 30% alcohol and extracting for 30 min,extraction temperature 70°C. Under the optimal conditions, the maximal yield of flavonoids was of 1.81%.

Extraction Technology and Determination of Total Flavonoid in Magnolia Liliiflora Petals

Orthogonal experiment optimization of microwave-assisted extraction (MAE) conditions of total flavonoids from Magnolia liliflora petals with the factors of ethanol concentration, extraction time,microwave power,solid/liquid ratio on the yield of total flavonoids were studied. The results showed that optimum extracting conditions of total flavonoids from Magnolia liliflora were as follows: adding 1:10 g·mL-1 of 40% alcohol and extracting for 5 min,microwave irradiation power 464W, under the optimal conditions, the maximal yield of flavonoids was of 2.08%.

ULTRASOUND-ASSISTED EXTRACTION OF TOTAL FLAVONOIDS FROM LEAVES OF SYRINGA OBLATA LINDL.

Leaves of Syringa Oblata Lindl. possess some important biological activities, such as antibacterial, antiviral, anti-inflammatory and antioxidant activities. These biological properties are mainly attributed to total flavonoids content. In this paper, ultrasound-assisted extraction of total flavonoids from leaves of Syringa Oblata Lindl. was studied. Effects of several experimental parameters, such as concentration of extracting solvent, ratio of liquid to material, extraction temperature, and time of sonication on extraction efficiencies of total flavonoids were evaluated. The best extraction conditions were: 1 g plant sample with 20 mL of 50% ethanol, at 60 °C for 50 min, obtaining a yield of total flavonoids of 92.00 ± 0.87 mg/g of plant. The results indicated that high amounts of total flavonoids can be extracted from leaves of Syringa Oblata Lindl. by ultrasound-assisted extraction technology.

English

The conditions for extraction of the total flavonoids from Flos pueraria was optimized and the purification and separation process were conducted to identify the main constituents of the total flavonoids as well. The solvent for extraction and its concentration, the solid-to-liquid ratio, extraction duration, temperature and ultrasonic frequency were investigated through a single-factor experiment. An orthogonal design (L9 (34) was constructed to achieve the best extraction conditions. The crude extract was then purified sequentially by petroleum ether, ethanol and chloroform, n-butyl alcohol, and eluted gradually with mixed mobile phase of methanol-chloroform solution in the silica gel column system. The ingredients were further separated by color reaction, ultraviolet spectrophotometry, high performance liquid chromatography, infrared and mass spectral analysis. The optimum extraction condition for the total flavonoids from F. pueraria was as follows: extraction  by 50% (v/v) methanol solution, the solid-to-liquid ratio at 1:30, extraction duration 2.0 h, the temperature of 70°C, ultrasound 3 times and 30 min each time. Five isoflavones were separated and identified as irisolidone, genistein, daidzein, kakkalide and puerarin, respectively. Under these optimal conditions, the yield of total flavonoids in the extracts was up to 17.5%, while the output rate previously reported is about 8% in general. Our study provided means for further development and utilization of the bioactive components from F. pueraria. Key words: Flos pueraria, flavonoids, orthogonal design, purification, ultrasound-assisted extraction.

Optimization of Ultrasonic-Assisted Extraction of Flavonoids from Fenugreek Using Response Surface Methodology and High-Performance Liquid Chromatography- Diode Array Detector Coupled with Mass Spectrometry Analysis

In this study, a central composite design based on response surface methodology was used to optimize ultrasonic-assistant extraction of flavonoids from Trigonella foenum-graecum L. The optimal conditions to obtain the highest yield of total flavonoids were as follows: ultrasonic time of 80 min, temperature of 48 degrees C and liquid to solid ratio of 20 g/mL, where 1.03 % yield was predicted. After extraction process optimization, a method for the analysis of flavonoids in Trigonella foenum-graecum L. by HPLC-DAD-MS was developed. The LOD and LOQ of these 4 compounds range from 50.94 to 70.31 ng/mL and 179.63 to 208.85 ng/mL, respectively. The established HPLC-DAD-MS method was proven to be useful for the quantitative and qualitative analysis of flavonoid constituents in fenugreek seed. This is the first report on preparation, identification and determination of flavonoids in fenugreek seed by different method and which would provide a basis for medicinal and edible use of fenugreek.

Ultrasonically Assisted Simultaneous Extraction of Isoorientin, Orientin, and Vitexin from Leaves of Neosinocalamus affinis (Rendle) Keng f. (N. affinis)

Ultrasonically assisted extraction (UAE) technique was developed for the simultaneous extraction of three main flavonoids, that is, isoorientin, orientin, and vitexin, from bamboo leaves of N. affinis, a discarded waste in the locality in this present work. Extraction conditions for each compound were optimized by factorial design coupled with exploration of interaction among factors using selectivity and yield of total flavonoids as evaluation indexes. The optimum conditions for UAE of these flavonoids were obtained. Recovery test and repeatability analysis of UAE technique under the optimized conditions was proven to be satisfactory for flavonoids investigated. Compared with the conventional refluxing extraction, ultrasonic method produces higher yields for each of flavonoids investigated and total flavonoids, with comparable selectivity for three flavonoids. Highly effective extraction of ultrasonic technique for flavonoids indicated it application prospect in the extraction of topic plant.

Improving Flavonoid Extraction from Ginkgo biloba Leaves by Prefermentation Processing

This paper presents a prefermentation treatment method involving fungi to improve flavonoid extraction from the leaves of Ginkgo biloba . The fungi employed for this treatment were screened from the soil present under an ancient ginkgo tree. Seventy-six strains belonging to 23 genera were isolated and identified by a molecular identification method employing 18S rDNA sequences. Thirty-three strains grew well using ginkgo leaves as the growth medium. One strain, Gyx086, with higher extracted yield of flavonoids and more similar to the control, was finally selected for prefermentation processing. The major fermentation factors were optimized by response surface methodology. The optimal conditions for the highest total falvonoid yield were 27.8 °C for temperature, 64.2% for moisture content, and 61 h for fermentation time. Under the optimal condition, a actual total flavonoid yield of 27.59 ± 0.52 mg/g dry weight culture sample was obtained, which was about 70% higher than that of unfermented gingko leaf samples.

Optimization of Ultrasonic Extraction of Total Flavonoids from Litchi (<i>Litchi chinensis</i> Sonn.) Seed by Response Surface Methodology

The ultrasonic technology was applied to optimize the processing parameters of extraction of flavonoids from litchi seed using the response surface methodology. The Box–Behnken design was used to analyze the effect of independent variables including extraction temperature (X1), methanol concentration (X2) and extraction time (X3) on the yield of total flavonoids as the dependent variable(Y). The analysis of variance (ANOVA) indicated that three independent variables, their quadratic terms and interaction terms showed a significant effect on yield of total flavonoids (YTF) except the interaction between X1and X2. A mathematical model with high determination coefficient was constructed, according to which the optimal extraction conditions were determined as follows: extraction temperature 70°C, methanol concentration 68%, extraction time 70 min. Under the above-mentioned condition, the experimental value of YTF was 3.55 g catechin equivalent per 100 g of dry (g CE/100 g ), which agreed with the predicted value perfectly, demonstrating the response surface model was suitable in optimizing the extraction of flavonoids from litchi seed.

Orthogonal test design for optimizing the extraction of total flavonoids from Inula helenium

Background:Inula helenium, which belongs to thecomposite family, is an important crude drug in traditional Chinese medicine.Materials and Methods:The effects of ethanol concentration, liquid to solid ratio, extraction temperature, and duration of microwave irradiation on the flavonoid extraction yield were studied through a single-factor experiment. An orthogonal array (L9(34)) was then constructed to achieve the best extraction conditions.Results:Variance analysis revealed that ethanol concentration significantly affected the extraction yield. The optimal conditions were as follows: ethanol concentration, 50% (v/v); liquid to solid ratio, 15:1; duration of microwave irradiation, 240 s; and extraction temperature, 60°C.Conclusion:Under these optimal conditions, the total flavonoid yield was 18.34 ± 0.64 mg/g. The use of a microwave-assisted process dramatically reduced the time needed for extraction of flavonoids from I. helenium.

Optimization of total flavonoid compound extraction from camellia sinesis using the artificial neural network and response surface methodology

The aim of this paper was to model and optimize the process of total flavonoid extraction from the green tea using the artificial neural network and response surface methodology, as well as the comparation of these optimization techniques. The extraction time, ethanol concentration and solid-to-liquid ratio were identified as the independent variables, while the yield of total flavonoid was selected as the dependent variable. Central composite design (CCD), using second-order polynomial model and multilayer perceptron (MLP) were used for fitting the obtained experimental data. The values of root mean square error, cross-validated correlation coefficient and normal correlation coefficient for both models indicate that the artificial neural network is better in prediction of total flavonoid yield than CCD. The optimal conditions using the desirability function at CCD model was achieved for the extraction time of 32.5 min, ethanol concentration of 100% (v/v) and solid-to-liquid ratio of 1:32.5 (m/v). The predicted yield at these conditions was 2.11 g/100 g of the dried extract (d.e.), while the experimentally obtained was 2.39 g/100 g d.e. The extraction process was optimized by the use of simplex method at MLP model. The optimal value of total flavonoid yield (2.80 g/100 g d.e.) was achieved after the extraction time of 27.2 min using ethanol concentration of 100% (v/v) at solid-to-liquid ratio of 1:20.7 (m/v). The predicted value of response under optimal conditions for MLP model was also experimentally confirmed (2.71 g/100 g d.e.).

Production of flavonoids and polysaccharide by adding elicitor in different cellular cultivation processes of Glycyrrhiza uralensis Fisch

In this study, callus and cell suspension were induced from seedlings of licorice (G. uralensis). In addition, it was revealed that the appropriate concentration of sucrose could promote the callus growth and increase the content of polysaccharide. The methyl jasmonate (MJ) and phenylalanine (PHE) could enhance the callus growth and content of flavonoids for G. uralensis. For producing more flavonoids and polysaccharide, two-stage cultivation was performed. In the first step, 30 g L−1 sucrose was fed into a 5-L balloon-type bubble bioreactor on 8th day of culture to enhance cell production and metabolite production. In a two-stage cultivation process, PHE (2 mM) and MJ (5 mg L−1) were added into a 5-L balloon-type bubble bioreactor after 10 days of culture. Using a fed-batch cultivation strategy (30 g L−1 sucrose was fed into a 5-L balloon-type bubble bioreactor on 8th day), polysaccharide production was enhanced to 1.19 g L−1, which was 2.12-fold greater than that in batch cultivation. The flavonoids yield (55.42 mg L−1) which was about 22 % higher than that in batch cultivation was obtained on 21st day. In a two-stage cultivation process, the polysaccharide content was increased by 1.14- and 2.12-fold compared with fed-batch cultivation and batch cultivation on 15th day. Meanwhile, total flavonoids yield (132.36 mg L−1) on 15th day, was increased by 2.26- and 2.67-fold compared with fed-batch cultivation and batch cultivation. In conclusion, two-stage cultivation process combined with the sucrose and elicitor treatment could promote both the callus growth and the secondary metabolites accumulation.

Optimization of total flavonoids content extracted from Flos Populi using response surface methodology

Flos Populi is an excellent source of total flavonoids used as an antidiarrheal drug. For better utilization of the resource, Box–Behnken design was used to optimize the extraction conditions of total flavonoids from Flos Populi. The effects of four independent variables in terms of extraction time, temperature, ethanol concentration, and the ratio of liquid to material on the flavonoids yield were determined and the optimal conditions for flavonoids were evaluated by means of response surface methodology. Correlation analysis of the mathematical regression model indicated that a quadratic polynomial model could be employed to characterize extraction process for the total flavonoids. Response surface plots showed that these independent variables significantly influenced the extraction yield of total flavonoids. The optimal extraction parameters to obtain the highest total flavonoids yield were temperature of 94.66°C, ethanol concentration of 45.92%, the ratio of liquid to material of 18.55:1 (mL/g), and time duration of 2.27h. The average experimental total flavonoids yield under the optimum conditions was found to be 53.08±0.43mg/g, which agreed with the predicted value of 52.04mg/g. The extraction method was applied successfully to extract total flavonoids from Flos Populi.

Optimization of Microwave Assisted Extraction Conditions for Total Flavonoids in Toona Sinensis Leaves Using Response Surface Methodology

Abstract In this study Microwave assisted extraction (MAE) was employed to extract total flavonoids (TF) from the Toona sinensis leaves (TSL). Single factor experiments to evaluate the effects of the process parameters (extraction duration, solvent-solid ratio, ethanol volume fraction, and extraction times) on TF yield of TSL was performed, and response surface methodology (RSM) were used to optimize the extraction parameters. The analysis of variance indicated that the linear coefficients extraction duration, the quadratic terms of extraction duration and solvent-solid ratio, had highly significant effects on the TF yield (P<0.01). And, the quadratic terms of ethanol volume fraction had significant effects on the TF yield (P<0.05). The optimal conditions to obtain the maximum TF yield from TSL were extraction duration 85s, solvent-solid ratio (v/w, mL/g) 20, ethanol volume fraction 50%, microwave power 700 W, and repeated extraction two times. Under these optimal conditions, the TF yield was up to 2.39%. The analysis of variance (ANOVA) and validation test indicated that the method is useful and reliable.

Effect of shading, nitrogen and magnesium fertilizer on phyllanthin and total flavonoid yield of Phyllanthus niruri in Indonesia soil

Phyllanthus niruri is widely used as traditional medicine especially in Indonesia. The need to produce a standardized herbal medicine raw material has urged a research to study the effect of shading, nitrogen (N) and magnesium (Mg) fertilizer dosage on phyllanthin and total flavonoid yield of P. niruri. Pot experiment was conducted by using Inceptisols soil from Yogyakarta, Indonesia. The study was carried out in a split plot design with three replications. The main plot was shaded with 3 levels (0, 60 and 95%). The subplots was urea and magnesium suphate (MgSO4) fertilizer (proportion 5 : 1) with 5 levels (0, 1+0.2, 2+0.4, 3+0.6 and 4+0.8). The period of harvesting were 1, 2 and 3 months after planting. The results indicated that application of 3 g urea, 0.6 g MgSO4 and 60% paranets shading could improve the plant growth, N-Mg uptake, total flavonoid and phyllanthin yield of P.niruri. Based on the regression curve, the optimum dosage that produced the highest total flavonoid yield was 2.68 g of urea and 0.54 g of MgSO4, while for obtaining the highest phyllanthin yield was 2.39 g of urea and 0.48 g of MgSO4 per plant. N and Mg absorption were positively correlated with total flavonoid and phyllanthin yield. The best period of harvest to obtain the highest total flavonoid and phyllanthin yield was 3 months after planting (plant age ± 4 months). Key words: Phyllanthus niruri, shading, nitrogen (N) and magnesium (Mg) fertilizer, phyllanthin, total flavonoid

Response surface optimized ultrasonic-assisted extraction of flavonoids from Sparganii rhizoma and evaluation of their in vitro antioxidant activities.

An efficient ultrasound-assisted extraction technique was employed to extract total flavonoids from Sparganii rhizoma. The optimum extraction conditions for the highest yield of total flavonoids were ethanol concentration 53.62%, ultrasonication time 29.41 min and ultrasound power 300 W, which were determined using response surface methodology. The extraction yields of the optimal ultrasound-assisted extraction were higher than using conventional extraction. The crude extract was then purified on a polyamide resin, whereby the flavonoids content in the purified extract increased to 94.62%. The antioxidant activities of the purified flavonoids including DPPH radical scavenging activity, ABTS+ radical scavenging activity, reducing power, hydroxyl radical scavenging activity and superoxide anion scavenging activity, were evaluated in vitro, which suggested that the flavonoids showed significant antioxidant activities. Rutin, kaempferol and formononetin were identified in the extract by comparing relative retention times and UV-Vis spectra with those of reference standards.

Growth, physiological characteristics and total flavonoid content of Glechoma longituba in response to water stress

The dried over-ground parts of the plant Glechoma longituba are one of the important ingredients used in traditional Chinese medicine (TCM). In this study, five different watering treatments, namely 95 to100% (T1), 80 to 85% (T2), 65 to 70% (T3), 50 to 55% (T4) and 35 to 40% (T5) of field capacity were used to investigate the effects of water deficiency on the growth, physiological characteristics and total flavonoid content of G. longituba. The study found that epidermis, palisade tissue, spongy tissue and vascular cylinder thickness peaked in the T2 treatment and then decreased in the other treatments in anatomy structure of leaf. And anatomical structure analysis of the stem revealed that the thickness of the cortex and the vascular cylinder were the highest in the T2 treatment, too. Water stress increased the thickness of the vessel walls in the xylem and the size of the parenchyma cells in the cortex, whereas it decreased the size of the parenchyma cells in the pith. As the water stress intensified, the resulting changes found in physiological indices suggested that water stress stimulated the level of redox activity. In addition, the yield of total flavonoids in the T2 treatment was higher than in the other treatments. Our results suggested that 80 to 85% of field capacity is the most suitable watering treatment for the growth of G. longituba. This procedure would be suitable for the domestication of G. longituba. Key words: Glechoma longituba, water treatment, physiological characteristics, total flavonoids.