Extraction of flavonoids in Brassica juncea and their hydroxyl radical scavenging effect of
【Abstract】 Objective To investigate the mustard in the extraction of total flavonoids, identification method and scavenging of hydroxyl radicals in order to make full use of
mustard plant resources and avoid waste of resources. Methods ultrasonic extraction of ethanol extracted from the mustard flavonoid substances, a flavonoid extracted to verify, and concentration was determined by spectrophotometry, using
mustard flavonoids on hydroxyl radical scavenging experiment. The results measured by the total flavonoid content of the sample C = 0.4450 mg / ml, the recovery rate was 100.7%, the purity and yield are high. Conclusion This method uses the
whole physical process without any pollution, the extraction of flavones from mustard in an effective way. Mustard flavonoids extract of OH radicals generated by Fenton system, a very good scavenging effect.
Key words ultrasonic extraction of total flavonoids
identification of radical scavenging mustard
Extraction of Total Flavone from Bragica juncea Coss and Its Effects on Scavenging of Hydroxyl Radicals
Abstract: ObjectiveTo make use of the resources of Bragica juncea coss, avoid wasting and approach the extraction of total flavanone of Bragica juncea Coss and its effects on scavenging of hydroxyl radicals. MethodsThe flavonoids were extracted by ethanol as the solvent from Bragica juncea coss with ultrasonic wave and ethanol extraction. Using
spectrophotometry to extract and check the flavone of Bragica
juncea Coss.Results The effect of extracts of Bragica juncea Coss scavenging of hydroxyl radicals was studied. By this method getting the density of the total flavone of Bragica
juncea Coss was C = 0.4450mg/ml and the rate of recovery was 100.7%. The outcome and the purity of the flavone were all very high. ConclusionThis method is a purely physical process and has not any pollution. It is an ideal way to extract the flavone of Bragica juncea coss. Total flavone from Bragica juncea Coss has effects on scavenging on hydroxyl radicals.
Key words: Ultrasonic wave treatment; Bragica juncea Coss; Total flavone; Identification; Hydroxyl radical; Scavenging
Mustard Bragica juncea Coss, also known as mustard,
potherb mustard, wrinkled leaf mustard, yellow mustard, for cruciferous plants mustard Brassia juncea (L.) Czern.et Coss oil mustard Brassia juncea (L.) Czern.et Coss.Var.gracilis Tsen et Lee's Spear leaves. Are cultivated all over the
country. Wash leaves in spring mining, civil frequently used to do pickles. Mustard sexual Weixin, Wen. Go lung, stomach, kidney. Li Huo Tan with gas, temperature in the spleen, dispelling wind scattered the blood, reduces swelling, pain and other effects. Indications Hantan cough, Xiongge unfavorable Wei Han deprivation, diarrhea, flu cold and bone injury, joint pain embolism . There are currently studies have shown that : Mustard contains flavonoid glycosides, acid resin, tannins, etc., including flavonoids and other
active ingredients for treating coronary heart disease, senile dementia, cerebral thrombosis, nervous system diseases and eliminate free radicals, antimicrobial, anti-cancer and so a
significant effect, non-toxic side effects, and thus developed
a variety of medicines and health food products.
Current home and abroad for mustard Flavonoids relative lack of study. Ethanol extraction is currently useful for extracting flavonoids mustard in a report , however, the relatively long time to extract, process more complex, extraction efficiency is low. In recent years, ultrasound technology, the extraction of plant alkaloids, glycosides, biologically active substances, animal tissue study of pulp and other toxic substances have been reported [4,5],
indicating that its has a low power consumption, high efficiency, non - destruction of the characteristics of the
active ingredient. Many studies have shown that the use of ultrasonic vibrations generated by a strong, high acceleration, strong cavitation, mixing function, etc., can accelerate the active ingredients of plant material into the solvent, thus increasing the extraction rate of the active ingredients, shortening extraction time, and also can avoid the impact of high temperature on the extraction composition , but the ultrasonic technology to extract the mustard in flavonoid research has not been reported. This paper reports the mustard with the ultrasonic extraction of total flavonoids, identify and extract flavonoids on hydroxyl
radical scavenging effect.
An instrument and reagent
1.1 Instrument KQ5200DB NC Ultrasonic Cleaner (Ultrasonic frequency 40 kHz, Kunshan City, Jiangsu Province, Ultrasonic Instrument Co., Ltd.); UV-755B UV-Vis spectrophotometer
(Shanghai Analytical Instruments Factory); ZF-I-type three -
UV analyzer (Shanghai Optical Instrument Factory Gucun); SHB-
III Multi-purpose circulating water pump (Zhengzhou Great Wall Industry and Trade Branch Co., Ltd.).
1.2 Reagents 95% ethanol, AR; sodium nitrite AR; aluminum
nitrate AR; sodium hydroxide AR; three aluminum chloride AR; hydrochloride, AR; ammonia AR; glacial acetic acid AR; Ethyl acetate AR; acid magnesium AR; magnesium powder AR; n-butanol
AR; rutin standard (Chinese medicines and biological
products); hydrogen peroxide AR; ferrous sulfate AR; acid AR; mustard, were collected from Guangxi Baise City.
2 Methods and Results
2.1 Extraction of total flavonoids to take fresh mustard, drying, crushing. Weigh mustard powder 6 g, add 80 ml 95%
ethanol, using ultrasonic extraction 2.5 h, leaching. Residues together with 80 ml 95% ethanol, using ultrasonic extraction 2.5 h, leaching, merging filtrate, vacuum recovery of ethanol to the filtrate until the remaining 25 ml, placed in 250 ml
flask, with 60% ethanol was diluted to scale, may sample fluid.
2.2 Determination of rutin as reference substance based on determination of total flavonoids content of mustard, join
the aluminum ion reagent, while controlling suitable pH value,
and the flavonoid compounds and aluminum salts to form complexes, in the visible region can get stable characteristic absorption peaks .
2.3 quantitative test - Determination of total flavonoids
2.3.1 Wavelength selection to take samples of fluid
amount, in 0.30 ml 5% sodium nitrite solution under alkaline conditions exist, after aluminum nitrate color, with its reagent blank reference solution in the wavelength range 420 ~ 700 nm measured network compounds of the absorbance at 510 nm complex with maximum absorption wavelength, so when you use the wavelength of measurement.
2.3.2 Standard curve drawing lessons were precise control fluid rutin (0.10 mg / ml) 0.00,0.50,1.00,2.00,3.00,4.00,5.00 ml flask at 10.00 ml, respectively of 5% sodium nitrite
solution, 0.30 ml, shaken, put it aside for 6 min; plus 10% aluminum nitrate solution, 0.30 ml, shaken, put it aside for 6 min; plus 4% sodium hydroxide solution 4.00 ml, with 60% ethanol was diluted to scale, shake, put it aside for 12 min,
with reagent blank as reference solution, measured at absorbance at 510 nm. The results in Table 1. Table 1 UV-Vis
spectrophotometer determination of rutin in control solution absorbance results (omitted)
According to Table 1 were the regression equation: A =-
0.0221 13.7578C, r = 0.9999.
2.3.3 Determination of precision extract fluid samples absorb 1.00 ml, owned 10.00 ml flask, according to the standard curve preparation method of determination of absorbance, A = 0.368, according to the regression equation: A
=- 0.022 1 13.757 8 C, r = 0.999 9 sample calculation of total flavonoid content of C = 0.445 0 mg / ml.
2.3.4 Experimental precision the amount of recovery to take samples of liquid 1.00 ml, by adding the standard rutin reference substance 1.00 ml (0.10 mg / ml), with the former samples were measured absorbance measurement operation, find the recovery rate was 100.7%. Reposted elsewhere in the paper for free download http://
2.4 Qualitative test - Identification of flavonoids
2.4.1 color reaction
UV-light-color-forming reaction: take the sample solution points in filter paper, in the visible light greyish-yellow
under UV light was gray-brown and a fluorescent spots.
Concentrated ammonia reaction: take the sample solution
points in the filter paper on top of the filter paper in the ammonia smoked 30 s, immediately observed under ultraviolet light, showing a very obvious brown fluorescent spots.
Of aluminum chloride reaction: take sample solution point
in the filter paper, dropping 1% ethanol solution of aluminum chloride, drying. In the visible light greyish-yellow
fluorescence under UV light yellow spots.
Of magnesium acetate reaction: take sample solution point in the filter paper, dropping 1% methanol solution of
magnesium acetate, drying, ultraviolet light yellow spots.
Hydrochloric acid - Mg reaction: Take 1ml of ethanol
extract in the test tube, add magnesium powder, then add a few drops of concentrated hydrochloric acid (1 accession), the
bubble was at purple.
2.4.2 Paper Chromatography check points in 10 μl of
sample solution on the filter paper. With n-butanol: acetic
acid: water = 4:1:5 (volume ratio) as the agent, started up 5 h, removed dry. Spray 1% ethanol solution of aluminum
chloride. After drying under ultraviolet light, visible fluorescent spots.
2.5 total flavonoids extract on scavenging OH reference method of Fenton reaction to the establishment of the reaction system model , the use of H2O2 and Fe2 mixing OH, but OH is
highly reactive, the survival time is short, if the reaction system by adding salicylic acid, can effectively capture the OH, and produce non-ferrous products. Reactive as follows: H2O2 Fe2 ? Fe3 * OH OH -
The products in a strong absorption at 510 nm, if in this reaction system by adding a clear OH function of EUT will
compete with the salicylic acid OH, thereby reducing the amount of colored product is generated using a fixed reaction time method , in the same volume of reaction system (8.8 mmol
/ L H2O2 1 ml, 9 mmol / L Fe2 l ml, 9 mmol / L 1 ml salicylic acid - ethanol solution) by adding a series of different concentrations of flavonoids extract of mustard, and with distilled water as reference, and the reagent blank solution,
comparing measured at 510 nm absorbance of different concentrations will be able to determine the measured object (flavonoids) on OH scavenging, its clearance rate formula: clearance rate ( %) = A0-AxA0 × 100%
In order to extract the experiment as the samples were
collected from samples of varying sizes were measured by constant volume to 10m1 of its OH scavenging effect. As can be seen from Figure 1, mustard extract flavonoids produced by the Fenton system, removal of OH have a certain role, as the
mustard total flavone concentration increased, the OH radical scavenging ability increased, indicating When the concentration of flavonoids increased, its clearance rate is also increased.
The experimental results show that this experimental
method is simple and recovery was 100.7%, to use this article provided by ultrasonic extraction and purification methods of flavonoids obtained from its high purity. Ultrasound can strengthen ethanol extraction method, to achieve time-saving,
high efficiency, energy-saving purposes, this method uses the whole physical process without any pollution, is an ideal way to extract flavonoid, has wide application prospects.
Flavonoids are a large class of natural products, widely exist in the plant kingdom, is the active ingredient in many Chinese herbal medicines. In nature is the most common flavonoids and flavonol Others include dihydrocodeine yellow (alcohol), isoflavones, double-flavone, flavanol, chalcone,
orange one, and the new anthocyanins and other flavonoids .
Natural source of bio-flavonoids molecular weight is small,
the body can be quickly absorbed through the blood-brain
barrier, can enter the adipose tissue, thus reflecting the following functions: to eliminate fatigue, protect blood
vessels, anti-arteriosclerosis, expand capillaries, and clear
the micro-circulation, anti - fat oxidation, anti-aging,
activation of cells in the brain and other organs function.
Flavonoids have anti-cancer, anti-tumor, anti-
cardiovascular and cerebrovascular diseases, anti-inflammatory
analgesic, immune regulation, lowering blood sugar, treatment of osteoporosis, antibacterial anti-virus, anti-oxidation,
anti-aging, anti-radiation effects [7, 8]. In recent years, the world set off the development boom of the botanicals,
botanical characteristics of its acclaimed line of natural and low toxicity, while its broad-spectrum flavonoids remarkable
pharmacological effects. The experimental results show that the more abundant mustard contains flavonoids, which extract
flavonoids on hydroxyl radical scavenging effect has a very good. Therefore, there is a wide range of development and utilization of value, worthy of comprehensive utilization, strengthen resource development.
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