Ion chromatography–inductively coupled plasma mass spectrometry determination of arsenic species in marine samples

Arsenic speciation analysis in marine samples was performed using ion chromatography (IC) with inductively coupled plasma mass spectrometry (ICP‐MS) detection. The separation of eight arsenic species, viz. arsenite, monomethyl arsonic acid, dimethylarsinic acid, arsenate, arsenobetaine, tetramethylarsine oxide, arsenocholine and tetramethylarsonium ion was achieved on a Dionex AS4A (weaker anion exchange column) by using a nitric acid pH gradient eluent (pH 3.3 to 1.3). The entire separation was accomplished in 12 min. The detection limits for the eight arsenic species by IC–ICP‐MS were in the range 0.03–1.6 µ g l^?1, based on 3σ of the blank response (n = 6). The repeatability and day‐to‐day reproducibility were calculated to be less than 10% (residual standard deviation) for all eight species. The method was validated by analyzing a certified reference material (DORM‐2, dogfish muscle) and then successfully applied to several marine samples, e.g. oyster, fish muscle, shrimp and marine algae. The low power microwave digestion was employed for the extraction of arsenic from seafood products. Copyright © 2004 John Wiley & Sons, Ltd.     

Determination of different arsenic species in food‐grade spirulina powder by ion chromatography combined with inductively coupled plasma mass spectrometry

A “two‐step” pressurized microwave‐assisted extraction method coupled with ion chromatography with inductively coupled plasma mass spectrometry for the determination of different arsenic species in spirulina samples was developed. The extraction method used H₂O₂/H₂O (1:5, v/v) as solvent to extract all arsenic species except arsenite, which was extracted by using water as solvent. The extraction method had a satisfactory recovery (>96%) and took a short time (20.0 min). With our method, all arsenic species in spirulina samples were completely separated and determined with recoveries of 84–105% and relative standard deviations of 2–4%. Food‐grade spirulina powder samples from seven provinces (Inner Mongolia, Zhejiang, Fujian, Hainan, Yunnan, Jiangsu, and Guangxi) in China were analyzed using the optimized protocol. Arsenate was detected at the concentration range of 170–394 ng/g in all the spirulina samples. Dimethylarsinic acid was detected at the concentration range of 32–839 ng/g in spirulina from above‐six provinces except Guangxi. Monomethylarsonic acid (67 ± 3 ng/g) was detected only in spirulina from Yunnan province. Arsenite was detected at the concentration range of 28–147 ng/g in spirulina from above five provinces except Hainan and Guangxi. Five unknown organic arsenic species were found in spirulina from above six provinces except Guangxi.     

Optimization of microwave‐assisted extraction for six inorganic and organic arsenic species in chicken tissues using response surface methodology

Response surface methodology was applied to optimize the parameters for microwave‐assisted extraction of six major inorganic and organic arsenic species (As(III), As(V), dimethyl arsenic acid, monomethyl arsenic acid, p‐arsanilic acid, and roxarsone) from chicken tissues, followed by detection using a high‐performance liquid chromatography with inductively coupled mass spectrometry detection method, which allows the simultaneous analysis of both inorganic and organic arsenic species in the extract in a single run. Effects of extraction medium, solution pH, liquid‐to‐solid ratio, and the temperature and time of microwave‐assisted extraction on the extraction of the targeted arsenic species were studied. The optimum microwave‐assisted extraction conditions were: 100 mg of chicken tissue, extracted by 5 mL of 22% v/v methanol, 90 mmol/L (NH₄)₂HPO₄, and 0.07% v/v trifluoroacetic acid (with pH adjusted to 10.0 by ammonium hydroxide solution), ramping for 10 min to 71°C, and holding for 11 min. The method has good extraction performance for total arsenic in the spiked and nonspiked chicken tissues (104.0 ± 13.8% and 91.6 ± 7.8%, respectively), except for the ones with arsenic contents close to the quantitation limits. Limits of quantitation (S/N = 10) for As(III), As(V), dimethyl arsenic acid, monomethyl arsenic acid, p‐arsanilic acid, and roxarsone in chicken tissues using this method were 0.012, 0.058, 0.039, 0.061, 0.102, and 0.240 mg/kg (dry weight), respectively.     

An environmentally friendly sodium dodecyl sulfate‐synergistic microwave‐assisted extraction followed by ultra‐high‐performance liquid chromatography with photodiode array method for the simultaneous extraction and determination of iridoids,phenylpropanoids, and lignans from Eucommiae Cortex

A simple and green sodium dodecyl sulfate‐synergistic microwave‐assisted extraction method was developed to extract and determine the iridoids, phenylpropanoids, and lignans in Eucommiae Cortex followed by ultra‐high‐performance liquid chromatography with photodiode array detection. The biodegradable solution (sodium dodecyl sulfate) was used as a promising alternative to organic solvents. The response surface methodology provided the optimum extraction conditions (2 mg/mL sodium dodecyl sulfate, 1100 W microwave power, and 6 min extraction time). The recoveries of three types of components ranged from 95.0 to 105% (RSDs < 5%). The intra‐ and inter‐day precision and accuracy were less than 3.40% and within the range of 97.1‐105%, respectively. Compared with other extraction methods, this newly established method was more efficient and environmental friendly. The results demonstrated that sodium dodecyl sulfate‐synergistic microwave‐assisted extraction followed by ultra‐high‐performance liquid chromatography with photodiode array method was applicable for the simultaneous extraction and determination of these three types of compounds for quality evaluation of Eucommiae Cortex.     

Simultaneous determination of organophosphorus pesticides in Chinese herbal medicines by microwave‐assisted extraction coupled with dispersive‐solid phase extraction and gas chromatography

A reliable, rapid and nontoxic analytical method was proposed for the simultaneous determination of 16 organophosphorus pesticides in Chinese herbal medicines. The pesticides were extracted by ethanol and the experimental variables, such as temperature, extraction time and volume of ethanol, were optimized through orthogonal array experimental design. Cleanup of extracts was performed with dispersive‐solid phase extraction using primary secondary amine as the sorbent. The determination of pesticides in the final extracts was carried out by gas chromatography–flame photometric detection. Under optimized conditions the obtained recoveries, except for isocarbophos, were in the range 73.8–123%, with relative standard deviations equal to or lower than 15.2% and limits of detection ranging from 0.001 to 0.009 mg/kg. Copyright © 2010 John Wiley & Sons, Ltd.     

Rapid determination of oxindole alkaloids in cat's claw by HPLC using ionic liquid‐based microwave‐assisted extraction and silica monolithic column

Cat's claw is a large woody vine with hook‐like thorns, and has been traditionally used to treat inflammatory disorders in South and Central America. In this study, a rapid, validated high‐performance liquid chromatographic (HPLC) method using a silica monolithic column was developed for the simultaneous determination of oxindole alkaloids, namely rhynchophylline, pteropodine, isomitraphylline and isopteropodine, in cat's claw. The ionic liquid‐based microwave‐assisted extraction (ILMAE), considered as an environmentally friendly and powerful tool, was first applied in the extraction of oxindole alkaloids. To optimize the HPLC method, the stationary phases, pH values of mobile phase and flow rates were investigated. The validated HPLC method using a Monolithic RP_18e column (100 × 4.6 mm) enables these analytes to be separated almost twice as fast as with a conventional particulate column (~16 vs ~30 min) with limits of quantification and detection of 0.5 and 0.15 μg/mL, respectively. The ILMAE conditions were optimized by the Taguchi orthogonal array design. In comparison with conventional water boiling extraction, ILMAE offers almost four times higher yields within an extremely short extraction time. The developed HPLC coupled with ILMAE method could be efficient and practical for rapid determination of oxindole alkaloids in cat's claw.     

Fractional factorial design based microwave‐assisted extraction for the determination of organophosphorus and organochlorine residues in tobacco by using gas chromatography—mass spectrometry

Sample preparation is often the main bottleneck in analyzing biological samples. Particularly, effective evaluation of sample preparation conditions usually involves multiple factors and tedious and time‐consuming experiments. In this study, fractional factorial design, specifically orthogonal array testing, was employed to screen and optimize multiple extraction parameters in concise but representative experiments. An efficient and sensitive method was developed to determine organophosphorus and organochlorine pesticide residues in tobacco, via microwave‐assisted extraction and gas chromatography coupled with mass spectrometry detection. With orthogonal array design, screening, and optimization tests were subsequently conducted to determine the range, impact rank, and possible interactions of extraction temperature, time, microwave power, additive salt, and additive water. Orthogonal array testing selectively reduces the size and cost of experiments and meanwhile provides more information compared to the traditional experimental design that optimizes one factor at a time. A good linear range (0.02–2.00 μg/mL), limits of detection (0.001–0.098 μg/mL), and recovery rates (70.4–107.1%) were demonstrated by spiking known concentrations of multiple pesticide standards in tobacco samples. The established method was then successfully applied to the determination of multipesticide residues in raw tobacco leaves and commercial cigarettes.     

Determination of arsenic species in oyster tissue by microwave‐assisted extraction and liquid chromatography–atomic fluorescence detection

A method for the determination of arsenic species in oyster tissue is established. The extraction of arsenic species is carried out by using low‐power microwaves. Quantitative extraction is obtained at a power of 40 W, and in 5 min, using the extracting agent methanol/water (1 + 1). The measurements are carried out using liquid chromatography–UV irradiation–hydride generation–atomic fluorescence detection (LC–UV–HG–AFS). Three arsenic species were detected in oyster tissue: arsenobetaine (AsBet) (87%), a probable arsenosugar (AsS) (4.9%), and dimethylarsinate (DMA) (4.7%). No influence of the clean‐up, the microwave field or the IR drying system on the stability of the arsenic compounds was observed. The extracts can be kept stable up to 3 days at 4 °C. The performance of the method is proved on fresh samples, as they are usually analysed in routine laboratories. Copyright © 2001 John Wiley & Sons, Ltd.     

Determination of plant growth regulators in pears by microwave‐assisted extraction and liquid chromatography with electrospray ionization mass spectrometry

A new method for the determination of six plant growth regulators, 3‐indolylacetic acid, 3‐indolepropionic acid, 2‐naphthoxyacetic acid, 2,4‐dicholrophenoxyacetic acid, 1‐naphthlcetic acid, and methyl naphthalene‐1‐acetate, in pears was established by liquid chromatography with electrospray ionization mass spectrometry. In this study, a microwave‐assisted extraction technique was first applied for the determination of plant growth regulators in fruit and three cleanup techniques were, respectively, investigated for the purification of pear samples. The chromatographic separation was performed on a Diamonsil C₁₈ column by using 0.01 mol/L formic acid/ammonium formate buffer solution (pH 3.5)/methanol (35:65, v/v) as the mobile phase with a flow rate of 0.7 mL/min in 1:1 split mode. The LODs ranged from 0.3 to 1.9 μg/kg. Under optimized conditions, the average recoveries (five replicates) for six plant growth regulators (spiked at 0.01, 0.05, and 0.5 mg/kg) ranged from 78.9 to 118.0%, and the RSDs were 1.4–10.3%.     

Enantioselective analysis of bambuterol in human plasma using microwave‐assisted chiral derivatization coupled with ultra high performance liquid chromatography and tandem mass spectrometry

In this study, an enantioselective analytical method based on microwave‐assisted chiral derivatization coupled with ultra high performance liquid chromatography and tandem mass spectrometry was developed for the determination of bambuterol enantiomers in human plasma. The chiral derivatization reaction was greatly accelerated by microwave irradiation. Under the optimized conditions, both the derivatization time and separation time on column was only 3 min, and the lower limit of quantification was 2.5 pg/mL. The recoveries were in the range of 90.1–93.0% without significant matrix effect. Compared with the conventional heating chiral derivatization, microwave‐assisted chiral derivatization obtained higher chiral derivatization yields with much shorter time due to the effect of microwave irradiation. Furthermore, the racemization during the derivatization reaction was systematically investigated. The results showed the concentration of acetic acid and the reaction time had significant effects on the racemization, which could be well controlled during microwave‐assisted chiral derivatization for the short reaction time. Finally, this novel approach was demonstrated by determining bambuterol in human plasma of a clinical pharmacokinetic study in eight healthy volunteers. On the basis of the results, microwave‐assisted chiral derivatization coupled with ultra high performance liquid chromatography and tandem mass spectrometry as a simple and effective enantioselective analysis technique for the determination of chiral drugs in complex biological samples showed great promise.     

固相萃取分离-电感耦合等离子体质谱法测定海产品中的无机砷

分别将N-（β氨乙基）-?-氨丙基三甲氧基硅烷(AEAPTMS)、3-巯丙基三甲氧基硅烷(MPTMS)与四乙氧基硅烷（TEOS)水解共聚,制备氨基\巯基键合的硅胶材料。将此材料作为固相萃取（SPE）小柱的填充材料,建立了固相萃取快速分离富集海产品样品中五价砷As（Ⅴ）和三价砷As（Ⅲ）电感耦合等离子体质谱法（ICP-MS）测定海产品中无机砷的方法。研究了固相萃取小柱对无机砷的吸附原理、性能和洗脱条件,在pH3～4范围内固相萃取小柱材料有良好的选择吸附性,利用2%硝酸可将As（Ⅴ）洗脱,利用2%硝酸+0.1mol/L KIO3可将As（Ⅲ）洗脱。实际样品检测的加标回收率在72～103%之间,方法实现了海产品样品中无机砷形态快速、方便、准确的检测。     

Study on the PEG‐based microwave‐assisted extraction of flavonoid compounds from persimmon leaves

A method for PEG‐based microwave‐assisted extraction (MAE) of flavonoid compounds from persimmon leaves has been successfully developed. The extraction efficiency of total flavonoid content was evaluated by the chromatographic peak areas of quercetin and kaempferol, which are two bioactive components typically found in persimmon leaves. The best combination of extraction parameters was obtained with response surface methodology. A microwave power of 525 W, liquid to solid ratio of 17:1 mL/g, and PEG aqueous solution concentration of 60% w/w were identified as the optimum parameters. Extraction dynamics analysis indicated that the quercetin, kaempferol, and total flavonoid contents were rising with increasing extraction time up to 20–25 min, from which point onwards they all decreased. Under the optimum conditions, quercetin, kaempferol, and total flavonoid contents obtained from the sample were 1.20 ± 0.05, 0.64 ± 0.11, and 16.90 ± 0.06 mg/g, respectively. Compared with ethanol‐based MAE, and ethanol‐based and PEG‐based ultrasonic‐assisted extractions, PEG‐based MAE had higher efficiency for the extraction of flavonoid compounds from persimmon leaves. Overall, PEG‐based MAE represents an efficient choice for the extraction of bioactive substances from traditional Chinese medicines.     

Determination of arsenic species in Solanum Lyratum Thunb using capillary electrophoresis with inductively coupled plasma mass spectrometry

A simple and highly efficient interface to couple capillary electrophoresis with inductively coupled plasma mass spectrometry by a microflow polyfluoroalkoxy nebulizer and a quadruple ion deflector was developed in this study. By using this interface, six arsenic species, including arsenite, arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, and arsenocholine, were baseline‐separated and determined in a single run within 11 min under the optimized separation conditions. The instrumental detection limit was in the range of 0.02–0.06 ng/mL for the six arsenic compounds. Repeatability expressed as the relative standard deviation (n = 5) of both migration time and peak area were better than 2.5 and 4.3% for six arsenic compounds. The proposed method, combined with a closed‐vessel microwave‐assisted extraction procedure, was successfully applied for the determination of arsenic species in the Solanum Lyratum Thunb samples from Anhui province in China with the relative standard deviations (n = 5) ≤4%, method detection limits of 0.2–0.6 ng As/g and a recovery of 98–104%. The experimental results showed that arsenobetaine was the main speciation of arsenic in the Solanum Lyratum Thunb samples from different provinces in China, with a concentration of 0.42–1.30 μg/g.     

Ultrasound/microwave‐assisted solid–liquid–solid dispersive extraction with high‐performance liquid chromatography coupled to tandem mass spectrometry for the determination of neonicotinoid insecticides in Dendrobium officinale

A one‐step ultrasound/microwave‐assisted solid–liquid–solid dispersive extraction procedure was used for the simultaneous determination of eight neonicotinoids (dinotefuran, nitenpyram, thiamethoxam, clothianidin, imidacloprid, acetamiprid, thiacloprid, imidaclothiz) in dried Dendrobium officinale by liquid chromatography combined with electrospray ionization triple quadrupole tandem mass spectrometry in multiple reaction monitoring mode. The samples were quickly extracted by acetonitrile and cleaned up by the mixed dispersing sorbents including primary secondary amine, C₁₈, and carbon‐GCB. Parameters that could influence the ultrasound/microwave‐assisted extraction efficiency such as microwave irradiation power, ultrasound irradiation power, temperature, and solvent were investigated. Recovery studies were performing well (70.4–113.7%) at three examined spiking levels (10, 50, and 100 μg/kg). Meanwhile, the limits of quantification for the neonicotinoids ranged from 0.87 to 1.92 μg/kg. The method showed good linearity in the concentration range of 1–100 μg/L with correlation coefficients >0.99. This quick and useful analytical method could provide a basis for monitoring neonicotinoid insecticide residues in herbs.     

Coupling nanoliter high‐performance liquid chromatography to inductively coupled plasma mass spectrometry for arsenic speciation

Nanoliter high‐performance liquid chromatography shows low consumption of solvents and samples, offering one of the best choices for arsenic speciation in precious samples in combination with inuctively coupled plasma mass spectrometry. A systematic investigation on coupling nanoliter high‐performance liquid chromatography to inductively coupled plasma mass spectrometry from instrument design to injected sample volume and mobile phase was performed in this study. Nanoflow mobile phase was delivered by flow splitting using a conventional high‐pressure pump with reuse of mobile phase waste. Dead volume was minimized to 60 nL for the sheathless interface based on the previously developed nanonebulizer. Capillary columns for nanoliter high‐performance liquid chromatography were found to be sensitive to sample loading volume. An apparent difference was also found between the mobile phases for nanoliter and conventional high‐performance liquid chromatography. Baseline separation of arsenite, arsenate, monomethylarsenic, and dimethylarsenic was achieved within 11 min on a 15 cm C₁₈ capillary column and within 12 min on a 25 cm strong anion exchange column. Detection limits of 0.9–1.8 μg/L were obtained with precisions variable in the range of 1.6–4.2%. A good agreement between determined and certified values of a certified reference material of human urine (GBW 09115) validated its accuracy along with good recoveries (87–102%).     

Rapid preparative separation of six bioactive compounds from Gentiana crassicaulis Duthie ex Burk. using microwave‐assisted extraction coupled with high‐speed counter‐current chromatography

A rapid method combining microwave‐assisted extraction (MAE) and high‐speed counter‐current chromatography (HSCCC) was applied for preparative separation of six bioactive compounds including loganic acid ( I ), isoorientin‐4′‐O‐glucoside ( II ), 6′‐O‐β‐d ‐glucopyranosyl gentiopicroside ( III ), swertiamarin ( IV ), gentiopicroside ( V ), sweroside ( VI ) from traditional Tibetan medicine Gentiana crassicaulis Duthie ex Burk. MAE parameters were predicted by central composite design response surface methodology. That is, 5.0 g dried roots of G. crassicaulis were extracted with 50 mL 57.5% aqueous ethanol under 630 W for 3.39 min. The extract (gentian total glycosides) was separated by HSCCC with n‐butanol/ethyl acetate/methanol/1% acetic acid water (7.5:0.5:0.5:3.5, v/v/v/v) using upper phase mobile in tail‐to‐head elution mode. 16.3, 8.8, 12., 25.1, 40.7, and 21.8 mg of compounds I–VI were obtained with high purities in one run from 500 mg of original sample. The purities and identities of separated components were confirmed using HPLC with photo diode array detection and quadrupole TOF‐MS and NMR spectroscopy. The study reveals that response surface methodology is convenient and highly predictive for optimizing extraction process, MAE coupled with HSCCC could be an expeditious method for extraction and separation of phytochemicals from ethnomedicine.     

Comparison of microwave‐assisted and conventional extraction of mangiferin from mango (Mangifera indica L.) leaves

Mangiferin is the main bioactive component in mango leaves, which possesses anti‐inflammatory, antioxidative, antidiabetic, immunomodulatory, and antitumor activities. In the present study, a microwave‐assisted extraction method was developed for the extraction of mangiferin from mango leaves. Some parameters such as ethanol concentration, liquid‐to‐solid ratio, microwave power, and extraction time were optimized by single‐factor experiments and response surface methodology. The optimal extraction conditions were 45% ethanol, liquid‐to‐solid ratio of 30:1 (mL/g), and extraction time of 123 s under microwave irradiation of 474 W. Under optimal conditions, the yield of mangiferin was 36.10 ± 0.72 mg/g, significantly higher than that of conventional extraction. The results obtained are beneficial for the full utilization of mango leaves and also indicate that microwave‐assisted extraction is a very useful method for extracting mangiferin from plant materials.     

Determination of seven arsenic species in seafood by ion exchange chromatography coupled to inductively coupled plasma-mass spectrometry following microwave assisted extraction: method validation and occurrence data

The determination of seven arsenic species in seafood was performed using ion exchange chromatography on an IonPac AS7 column with inductively coupled plasma mass spectrometry detection after microwave assisted extraction. The effect of five parameters on arsenic extraction recoveries was evaluated in certified reference materials. The recoveries of total arsenic and of arsenic species with the two best extraction media (100% H₂O and 80% aqueous MeOH) were generally similar in the five seafood certified reference materials considered. However, because MeOH co-elutes with arsenite, which would result in a positively biased arsenite concentration, the 100% H₂O extraction conditions were selected for validation of the method. Figures of merit (linearity, LOQs (0.019-0.075 mg As kg⁻¹), specificity, trueness (with recoveries between 82% (As(III)) and 104% (As(V) based on spikes or certified concentrations), repeatability (3-14%), and intermediate precision reproducibility (9-16%) of the proposed method were satisfactory for the determination of arsenite, monomethylarsonic acid, dimethylarsinic acid, arsenate, arsenobetaine and arsenocholine in fish and shellfish. The performance criteria for trimethylarsine oxide, however, were less satisfactory. The method was then applied to 65 different seafood samples. Arsenobetaine was the main species in all samples. The percentage of inorganic arsenic varied between 0.4-15.8% in shellfish and 0.5-1.9% at the utmost in fish. The main advantage of this method that uses only H₂O as an extractant and nitric acid as gradient eluent is its great compatibility with the long-term stability of both IEC separation and ICP-MS detection.     

Optimization of microwave‐assisted extraction followed by RP‐HPLC for the simultaneous determination of oleanolic acid and ursolic acid in the fruits of Chaenomeles sinensis

An optimized microwave‐assisted extraction (MAE) method and an efficient HPLC analysis method were developed for fast extraction and simultaneous determination of oleanolic acid and ursolic acid in the fruit of Chaenomeles sinensis. The open vessel MAE process was optimized by using a central composite experimental design. The optimal conditions identified were microwave power 600 W, temperature 52°C, solvent to material ratio 32 mL/g and extraction time 7 min. The results showed that MAE is a more rapid extraction method with higher yield and lower solvent consumption. The HPLC–photodiode array detection analysis method was validated to have good linearity, precision, reproduction and accuracy. Compared with conventional extraction and analysis methods, MAE–HPLC–photodiode array detection is a faster, convenient and appropriate method for determination of oleanolic acid and ursolic acid in the fruits of C. sinensis.     

Extraction and isolation of flavonoid glycosides from Flos Sophorae Immaturus using ultrasonic‐assisted extraction followed by high‐speed countercurrent chromatography

A method of ultrasonic‐assisted extraction followed by high‐speed countercurrent chromatography was established for the extraction and isolation of three flavonoid glycosides, i.e. rutin, narcissin, and nicotiflorin from Flos Sophorae Immaturus. The effects of ultrasonic‐assisted extraction factors for the main flavonoid compound (rutin) from Flos Sophorae Immaturus were optimized using Box–Behnken design combined with response surface methodology. The optimum conditions were determined as ultrasonic power 83% (600 W), solvent‐to‐material ratio 56:1, methanol concentration 82% v/v, and extraction time 60 min. Three bioactive flavonol glucosides, rutin, narcissin, and nicotiflorin were isolated from Flos Sophorae Immaturus using high‐speed countercurrent chromatography. The separation was performed with a two‐phase solvent system containing ethyl acetate/n‐butanol/methanol/water (4:0.9:0.2:5, v/v). Amounts of 87 mg of rutin, 10.8 mg of narcissin, and 1.8 mg of nicotiflorin were isolated from 302 mg of crude extract of Flos Sophorae Immaturus in a one‐step separation within 160 min with purities of 99.3, 98.0, and 95.1%, respectively, as determined by HPLC with diode array detection. Their structures were characterized by UV, MS, and NMR spectroscopy. It was demonstrated that the established method was simple, fast, and convenient, which was feasible to extract and isolate active flavonoid glycosides from Flos Sophorae Immaturus.