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Flavonoids are widely found in medicinal plants, which have important medical properties. Flavonoids were proved to have many pharmacological activities, such as anti-oxidation, antitumor, antimutation, anti-inflammatory, antibacterial and anti-aging. The extraction of flavonoids is the crucial link in their clinical applications. In recent years, many emerging Chinese medicine extraction methods have also been widely used in the extraction of flavonoids. This paper reviews the current application of new methods for flavonoid extraction, in order to provide references for the extraction, development and utilization of flavonoids. These new extraction methods include supercritical fluid extraction (SFE), ultrasonic assisted extraction (UAE), microwave assisted extraction (MAE), pressurized liquid extraction (PLE), pulsed electric field (PEF) assisted extraction, enzyme assisted extraction (EAE), green solvent extraction, steam explosion assisted extraction, dynamic high pressure microfluidization (DHPM) assisted extraction, etc.
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OBJECTIVE To establisha simultaneous determination method of three main alkaloids in Plumula nelumbinis by high performance liquid chromatography(HPLC), including liensinine, isoliensinine and neferine, and optinize the ultrasonic-assisted extraction process of total alkaloids from Plumula nelumbinis by using response surface methodology based on Box-Behnken design. METHODS The alkaloids were separated on a RP-HPLC C18 monolithic column(4.6 mm×50 mm, Merck, Darmstadt, German) with a mobile phase consisting of methanol-water-triethylamine-acetic acid(70∶30∶0.2∶0.05), and the method was validated for linearity, sensitivity and extraction recovery. Based on the single-factor analysis, a mathematical model was constructed to analyze the effects of each factor of the ultrasonic-assisted extraction and their interactions on alkaloids yields of Plumula nelumbinis. The three independent variables were volume fraction of ethanol(A), solid-liquid ratio(B) and ultrasound extraction time(C), respectively. RESULTS The HPLC method used to separate three alkaloids within 5 min showed an excellent linear correlation (r2>0.999) in the range of 3.4-340 μg·mL-1(liensinine), 3.47-347 μg·mL-1(isoliensinine) and 3.47-347 μg·mL-1 (neferine). The limits of detection(LODs) of three alkaloids were 0.17, 0.69 and 0.69 μg·mL-1, and the limits of quantification were 0.34, 1.73 and 1.73 μg·mL-1, respectively. The intra-day and inter-day variations of RSD were less than 5%, and the extraction recovery rang from 97.06% to 104.12% with RSD ranging from 0.08% to 0.87%(n=6). The optimum extraction conditions were: solid-liquid ratio, 1∶30.3; volume fraction of ethanol, 69.8%; ultrasound extraction time, 29.0 min. The yield under the optimum conditions was found to be(14.82±0.42) mg·g-1, which was agreed closely with the predicted value of 14.70 mg·g-1. CONCLUSION The HPLC method used to determine the three main alkaloids in Plumula nelumbinis shows ideal characteristics of quickness, accuracy, high sensitivity and good repeatability.The optimum ultrasonic extraction technology has expressed excellent extraction ability of alkaloids in Plumula nelumbinis which indicates the RSM result is reasonable and effective.
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OBJECTIVE:To optimize extraction technology of protein from Cryptotympana pustulata and investigate its in vitro antioxidant activity,so as to provide reference for further research of protein from C. pustulata. METHODS:Using extraction amount of protein as response value,based on single factor test,Box-Behnken response surface methodology was used to optimize the ratio of liquid to material,ultraonic time and extraction times.Validation test was conducted. Using Vitamin C(VC)as positive control, in vitro antioxidant activity of protein from C. pustulata was evaluated by using scavenging rate of 1, 1-diphenyl-2-picrylhydrazyl(DPPH)and 2,2′-azino-bis(3-ethyl benzothiazoline-6-sulfonic acid)(ABTS)free radical as index. RESULTS:The optimal extraction condition of protein from C.pustulata was as follows as the ratio of liquid to material 28:1(mL/g), ultrasonic time of 65 min,extracting for twice. In validation test,average extraction amount of protein from C. pustulata was 65.45 mg/g(RSD=1.68%,n=3),relative error of which to predicted value was 5.48%. The protein from C. pustulata showed strong scavenging effect on ABTS and DPPH free radicals. When the concentration of protein from C. slough was 0.2 mg/mL,and the scavenging rate of it to ABTS free radicals was 97%,the effect of which was similar to VC.The protein from C. pustulata showed weak scavenging ability to DPPH free radical,IC50was 0.96 mg/mL,the effect of which was not as good as VC. CONCLUSIONS:The extraction technology of protein from C. pustulata optimized by Box-Behnken response surface methodology shows high accuracy and good reliability.The protein from C.pustulata shows certain antioxidant activity in vitro.
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A green, rapid and efficient method was developed for the extraction of 4 kinds of hosenkosides from the seeds of Impatiens balsamina L. The hosenkosides were extracted by ionic liquid, and then enriched by solid phase extraction. The effects of the kinds and volume of ionic liquid, pH value, ultrasonic time, solid phase extractant and eluent on the extraction fields were investigated and optimized. Under the optimum conditions, the recoveries of 4 kinds of hosenkosides were between 92. 1％ and 108. 2％ , the correlation coefficients were 0. 9945-0. 9975, and the detection limits were 1. 8-4. 5 μg / mL. The RSD values were all less than 3. 9％ . The experimental results showed that this method was fast, efficient, environmental protection. This study provided a reference for the extraction and enrichment of trace components in Chinese medicinal herbs.
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A new ultrasonic-assisted extraction (UANE) method coupled with solid phase extraction (SPE) using ultrasonic fountain was established for the extraction of eight common ginsenosides from leaves of Panax quinquefolium L. The extraction system has been designed and several experimental parameters,including the type and volume of extraction solvent,pH value and salt concentration of extraction solvent,type and volume of elution solvent,and amount of C18, extraction time were examined and optimized. Under the optimal conditions,the recoveries of ginsenosides were in the range of 96. 3% -110. 6%, the relative standard deviations (RSDs) were in the range of 2.8%-4.3%,indicating that the method has a good performance for the extraction of these ginsenosides. Compared with traditional UANE-SPE method, the modified method simplified the extraction device,shortened the extraction time and improved the extraction efficiency.
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Using six kinds of ionic liquids as extractants, ultrasonic-assisted extraction coupled with HPLC method was developed for the simultaneous determination of wilforgine, wiforizine, triptophenolide, wilforine and triptoquinone A in Tripterygium hypoglaucum. The separation was performed on an Inertsil ODS-4 column with the mobile phase of acetonitrile-0.1% phosphoric acid in gradient elution at a flow rate of 0.75 mL•min⁻¹. Detection wavelength was 220 nm and the column temperature was 30℃. Under the optimal extractions, the results showed that triptophenolide and triptoquinone A had the highest extraction yield by using 0.6 mol•L⁻¹ [BMIm]PF6 methanol solution as extraction solvent with the solid-liquid ratio of 1∶10. The calibration curves of triptophenolide and triptoquinone A showed a good linearity in the range of 0.000 65-0.026, 0.066 55-2.662 μg (r=0.999 9)respectively. The average recovery was 102.4% and 97.90% with RSD of 2.5% and 1.5%, respectively. Wilforgine, wiforizine and wilforine had the highest extraction yield when using 0.6 mol• L⁻¹ [BMIm]PF6absolute ethanol solution as extraction solvent with the solid-liquid ratio of 1∶10. The content of wilforgine, wiforizine and wilforine from 0.023 9-0.956, 0.002 7-0.108, 0.006 4-0.256 μg showed a good linearity (r=0.999 9), and the average recovery was 100.6%,99.50% and 98.70% with RSD of 2.1%,1.9% and 2.7%, respectively. The results indicated that this method is convenient, reliable and green, and can be used as a reliableanalytical method for the quality control of T.hypoglaucum.
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OBJECTIVE: To determine the optimum ultrasonic-assisted extracting process of total flavonoids from Zingiber mioga by response surface methodology, and then study the antioxidant activity of the extract. METHODS: On the basis of single factor experiments, a 4-factor, 3-level Box-Behnken center-united experiment was conducted. The Box-Behnken experiment analyzed and optimized the processing conditions by response surface methodology, then the antioxidant activity of the extract was determined. RESULTS: The optimum conditions at ultrasonic power of 300 W were as follows treatment temperature 60℃, ethanol concentration 85.99%, liquid-to-solid ratio 98.08 mL·g-1, treatment time 15 min. Under the optimum conditions, the extraction rate of the total flavonoids was 37.01%, with a bias of 4.28% compared with the predicted value of 35.49%; the reducing power of the total flavonoids from Zingiber mioga at the concentration of 1.038 7 mg·mL-1 was equal to that of vitamin C of 56.60 μg·mL-1; the ·OH radical scavenging ratio of the total flavonoids from Zingiber mioga was 73.07%, the DPPH radical scavenging ratio was 65.72%, and the inhibition rate of anti-lipid peroxidation was 49.94%. CONCLUSION: The total flavonoids from Zingiber mioga can be developed as a new type of antioxidant agents, and regression analysis and parameter optimization of the extracting process can be conducted by using response surface methodology.
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OBJECTIVE@#To investigate suitable condition for extraction of the active components from Ajuga nipponensis (A. nipponensis).@*METHODS@#Orthogonal experimental design was used to determine the optimal extraction parameters for ecdysterones and flavonoids. Finally, the hepatoprotective abilities of A. nipponensis extracts were evaluated by CCl4-induced animal models.@*RESULTS@#Maximum yields of flavonoids (7.87 ± 0.10) mg/g and ecdysterones (0.73 ± 0.02) mg/g could be obtained when the extraction time was 50 min, the extraction temperature was 60 °C, and the ratio of sample to 70% (v/v) ethanol was 1:20 (w/w). The antioxidant property of A. nipponensis was correlated to the concentration of its extracts. At 5 mg/mL, A. nipponensis extract scavenged 84.8% of DPPH radical and had absorbance values of 2.43 ± 0.04 reducing power. Upon CCl4-induced liver injury, glutamic oxaloacetic transaminase and glutamic pyruvic transaminase decreased significantly after the mice were treated with A. nipponensis. Histological researches also explained that A. nipponensis reduced the extent of liver lesions induced by CCl4.@*CONCLUSIONS@#A. nipponensis exhibited potent antioxidant activity in chemical experimental models and hepatoprotective effect against CCl4-induced liver damage.
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Objective:To optimize the ultrasonic-assisted extraction technology of polysaccharide from Tremella by Box-Benhnken response surface methodology .Methods:The single-factor extraction tests and response surface analysis were employed to optimize the technology conditions , including ultrasonic power , ultrasonic time and liquid/solid ratio.Results:The experiment results indicated that the optimal extraction conditions were ultrasonic power of 360 W, ultrasonic time of 23 min and liquid/solid ratio of 45 ∶1 ( ml· g-1 ) . Under the conditions , 6 batches of polysaccharide from Tremella were extracted .The average extraction rate of Tremella polysaccharides was 20.4%±1.8%(n=6).Conclusion: The ultrasonic-assisted extraction process of Tremella polysaccharide using Box-Behnken response surface methodology is simple with good predictability .
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Objective To investigate suitable condition for extraction of the active components from Ajuga nipponensis (A. nipponensis). Methods Orthogonal experimental design was used to determine the optimal extraction parameters for ecdysterones and flavonoids. Finally, the hepatoprotective abilities of A. nipponensis extracts were evaluated by CCl
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OBJECTIVE: To optimize the ultrasonic-assisted extraction process of isoflavones from Pueraria lobata (Wild.) Ohwi using response surface methodology and measure the reducing power. METHODS: On the basis of single factor experiments, a four factors and five levels Box-Behnken experiment was designed. Then the reducing power of isoflavones was measured. RESULTS: The optimum conditions for the ultrasonic-assisted extraction process of isoflavone from Pueraria lobata (Wild.) Ohwi at ultrasonic power of 300 W were as follows: ethanol concentration 75%, liquid-solid ratio 35 mL · g-1, extraction time 35 min, and treatment temperature 50℃. Under the optimum conditions, the yield of polysaccharide from Pueraria lobata (Wild.) Ohwi was 24.18%, and the relative error between experimental value and predicted value was only 3.02%; the EC50 of the reducing power of isoflavones reached 0.135 mg · mL-1. CONCLUSION: The response surface methodology is suitable for regression analysis and optimization of the ultrasonicassisted extraction of polysaccharide from Pueraria lobata (Wild.) Ohwi.
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OBJECTIVE:To optimize ultrasonic-assisted extraction technology of total flavonoids from Vaccinium uliginosum. METHODS:Based on single factor test,the ultrasonic-assisted extraction technology of total flavonoids from V. uliginosum was op-timized by orthogonal test,using the yield of total flavonoids as index,with ethanol volume fraction,solid-liquid ratio and ultrason-ic power as factors. The optimized technology was validated and compared with traditional ethanol reflux method(reflux extracting for 2 times,2 h each time). RESULTS:The optimal extraction technology was as follows as the solid-liquid ration of 1:25,70%ethanol,ultrasonic power of 250 W(40 min for ultrasonic time). In validation test,the yield of total flavonoids from V. uliginosum was 3.98%(RSD=0.41%,n=3),and the yield of total flavonoids was 2.25% by ethanol reflux method. CONCLUSIONS:The ultrasonic-assisted extraction is obviously superior to ethanol reflux method on extraction time and effects,and it is suitable for in-dustrialized production.
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OBJECTIVE:To optimize the ultrasonic-assisted extraction technology of total flavonoids from Tibetan Scindapsus aureu. METHODS:The single-factor test was adopted to investigate the effects of extraction time,extraction temperature,the vol-ume fraction of ethanol,the ratio of liquid to solid and power on the extraction rate of total flavonoids from Tibetan Scindapsus au-reu in the process of ultrasonic-assisted extraction,based on which,four-factor and five-level Box-Behnken central component de-sign was performed with unchanged ultrasonic power,and then response surface method was adopted to analyze the effects of the 4 factors and their interaction on the extraction rate of total flavonoids to determine the optimal factor and level,and verification tests were conducted.RESULTS:The optimal ultrasonic-assisted extraction technology was as follows as extraction time of 29 min,tem-perature of 44 ℃,volume fraction of ethanol of 78%,ratio of liquid to solid of 13∶1(ml/g). In three verification tests,the aver-age extraction rate of total flavonoids was 2.130%,with an relative error of 0.005% compared to the theoretical value. CONCLU-SIONS:The optimal technology is reasonable and feasible,and can be used for theultrasonic-assisted extraction of total flavonoids from Tibetan Scindapsus aureu.
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A method of simultaneous determination of the eleven phenol and aniline dyes in oxidative hair dyes by ultrasonic-assisted extraction and high performance liquid chromatography-tandem mass spectrometry was developed. The orthogonal and single-factor experiments were designed to and optimize the ultrasonic-assisted extraction conditions, and the samples were extracted using 10 mL of 5% methanol under the conditions of ascorbic acid as an antioxidant for 10 min. The gradient elution program and the electrospray ionization mode change were together used for the optimization of the measurements, and the determinations were completed by using the multi-reaction monitoring scan. The detection limits were 1. 15-9. 43 μg/g, the recoveries of spiked samples were 88. 0%-118. 1%. The method can be used to determine trace prohibited and restricted dyes in hair dyes.
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Objective To isolate and determine Aloin and Aloeemodin in Barbodos Aloe by ionic liquid-based ultrasonic-assisted extraction coupled with high performance liquid chromatography, with 1-butyl-3-methylimidazolium chloride ([BMIM]Br) solution as the extraction solvent. Methods The separation was performed on Phenomenex C18 column (250 mm×4.6 mm, 5 μm) with detection wavelength of 360 nm. The mobile phase was consisted of methanol-0.3% acetic acid solution (65∶35) with the flow rate of 0.80 mL/min, and the column temperature at 35 ℃. Results The calibration curves for Aloin and Aloeemodin were liner within 0.000 336-1.68 μg (r=0.999 96) and 0.000 608-3.04 μg (r=0.999 76), respectively. The limit of detection (LOD) was 0.05 06 ng/mL and 0.262 ng/mL, respectively. The average recovery was 95.99% and 95.80%, respectively. Conclusion The method is simple, rapid, accurate, sensitive, low cost and environment-friendly, thus it provides an effective means for assaying anthraquinones in Barbodos Aloe.
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Axillary osmidrosis caused by excessive secretion of apocrine glands, which causes an malodor and extreme social embarrassment. To remove the apocrine glands, various types of treatment methods for axillary osmidrosis have been introduced. They could be summarized as manual of instrumental subdermal shaving, ultrasonic assisted liposuction, Botox(R) injection, electrical probe, laser dessication, etc. Surgical removal of apocrine glands has been the favored treatment for the several decades, but such complications as long-term postoperative immobilization period, skin necrosis, hematoma, and visible scars have been frequent. Recently, for the purpose of reducing these complications, some operators have advocated other procedures such as axillary ultrasonic assisted liposuction, Botox(R) injection, laser dessication. From March 2002 to April 2003, we have corrected the axillary osmidrosis in 80 cases using the subdermal shaving(24 cases), ultrasonic assisted liposuction(20 cases), Botox(R) injection(10 cases), electrical probe(26 cases). Of these operation, manual dermal shaving had the lowest recurrence rate, but disadvantages of this procedure was 4-5cm axillary scar, moderate post operative complication, long immobilization period. Otherwise Ultrasonic assisted liposuction had many advantages which were simple operation technique, short operation time, early recovery of social activity, rare post operative complication, invisible scar, lower recurrence rate. In case of Botox(R) injection was also simple operation technique, but which was short term effect about 6 months so it was necessary to treat repetitive injection but relatively high recurrence rate in severe osmidrosis. In case of electrical probe was also simple operation technique, but which was vague treatment effect, need to repetitive operation and relatively high recurrence rate in severe osmidrosis. In comparative evaluation of these methods, ultrasonic assisted liposuction was considered better results to osmidrosis treatment than others method in many aspects.
Subject(s)
Apocrine Glands , Cicatrix , Desiccation , Hematoma , Immobilization , Lipectomy , Necrosis , Recurrence , Skin , UltrasonicsABSTRACT
OBJECTIVE: To study on the effect of different extracting methods on the content determination of polysaccharide from Caulis Dendrobii.METHODS: Polysaccharide was extracted from Caulis Dendrobii respectively by reflux extraction,improved reflux extraction,ultrasound wave extraction and ultrasonic-assisted thermal reflux extraction.The content of polysaccharide was determined by phenol-sulfuric acid method.RESULTS: The determined results using these methods were as follows:(13.473?0.634)%,(15.692?0.743)%,(11.87?0.536)% and (16.29?0.407)%.CONCLUSION: The content determination of polysaccharide from Caulis Dendrobii was associated with the extraction method.The polysaccharide from Caulis Dendrobii could be extracted more thoroughly and rapidly by ultrasonic-assisted extraction technology.Therefore this method could be used to pretreat the sample in content determination of Caulis Dendrobili polysaccharide.