Efficient Sample Preparation Technique Research Articles

Visualising the Emerging Platform of Using Microalgae as a Sustainable Bio-Factory for Healthy Lipid Production through Biocompatible AIE Probes.

Nowadays, a particular focus is using microalgae to get high-valued health beneficiary lipids. The precise localisation of the lipid droplets (LDs) and biochemical changes are crucial to portray the lipid production strategy in algae, but it requires an in vivo tool to rapidly visualise LD distribution. As a novel strategy, this study focuses on detecting lipid bioaccumulation in a green microalga, Chlamydomonas reinhardtii using the aggregation-induced emission (AIE) based probe, 2-DPAN (C24H18N2O). As the messenger molecule and stress biomarker, hydrogen peroxide (H2O2) activity was detected in lipid synthesis with the AIE probe, TPE-BO (C38H42B2O4). Distinctive LDs labelled with 2-DPAN have elucidated the lipid inducing conditions, where more health beneficiary α-linolenic acid has been produced. TPE-BO labelled H2O2 have clarified the involvement of H2O2 during lipid biogenesis. The co-staining procedure with traditional green BODIPY dye and red chlorophyll indicates that 2-DPAN is suitable for multicolour LD imaging. Compared with BODIPY, 2-DPAN was an efficient sample preparation technique without the washing procedure. Thus, 2-DPAN could improve traditional fluorescent probes currently used for lipid imaging. In addition, the rapid, wash-free, multicolour AIE-based in vivo probe in the study of LDs with 2-DPAN could advance the research of lipid production in microalgae.

Unraveling the drug distribution in brain enabled by MALDI MS imaging with laser-assisted chemical transfer

Accurate localization of central nervous system (CNS) drug distribution in the brain is quite challenging to matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI), owing to the ionization competition/suppression of highly abundant endogenous biomolecules and MALDI matrix. Herein, we developed a highly efficient sample preparation technique, laser-assisted chemical transfer (LACT), to enhance the detection sensitivity of CNS drugs in brain tissues. A focused diode laser source transilluminated the tissue slide coated with α-cyano-4-hydroxycinnamic acid, an optimal matrix to highly absorb the laser radiation at 405 nm, and a very thin-layer chemical film mainly containing drug molecule was transferred to the acceptor glass slide. Subsequently, MALDI MSI was performed on the chemical film without additional sample treatment. One major advantage of LACT is to minimize ionization competition/suppression from the tissue itself by removing abundant endogenous lipid and protein components. The superior performance of LACT led to the successful visualization of regional distribution patterns of 16 CNS drugs in the mouse brain. Furthermore, the dynamic spatial changes of risperidone and its metabolite were visualized over a 24-h period. Also, the brain-to-plasma (B/P) ratio could be obtained according to MALDI MSI results, providing an alternative means to assess brain penetration in drug discovery.

Recent advances in sample preparation technologies for analysis of harmful substances in aquatic products

水产品含有丰富的蛋白、维生素和多种微量元素,是人们摄取动物性蛋白质的重要来源之一,我国是世界上最大的水产品消费国,其质量安全问题一直备受关注。但水产样品基质复杂,有害物质的含量低,须对其进行分离富集后才能进行检测,传统的液-液萃取、固相萃取和快速固相分散萃取等样品前处理技术在水产品分析中得到广泛应用,同时针对一些挥发性和超痕量有害物质检测时,固相微萃取同样体现出巨大优势。这些样品前处理技术可以有效去除基体对分析对象的干扰,提高检测方法的灵敏度和准确度。根据目标分析物性质的不同,选择合适的样品前处理技术,是水产品中有害物质分析的关键步骤。该文以水产品中有害物的来源不同,将其分为3类:(1)水产品中环境污染物的分析;(2)养殖运输和加工过程中有害物的分析;(3)水产品中生物毒素的分析。以这3类有害物质的分析为主线,综述了近10年水产品中有害物质分析的样品前处理技术,包括液-液萃取、固相萃取、固相微萃取、快速固相分散萃取和磁性固相萃取等。此外,还对各种技术的优缺点进行了探讨,并对其未来发展方向进行了展望。

Magnetic solid-phase extraction of gingerols in ginger containing products

In this study, a graphene oxide/magnetite (GO-Fe3O4) nanocomposite was synthesized and used as a sorbent in the magnetic solid-phase extraction (MSPE) of gingerols from fresh ginger rhizomes, ginger extracts, commercial tea samples, ginger candies, thermogenic supplements, and tonic water. An MSPE method was developed, and the main influencing parameters in the sample preparation process were investigated. After GO-Fe3O4 based MSPE, 6-gingerol, 8-gingerol, and 10-gingerol were determined by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The whole GO-Fe3O4-MSPE-LC-MS/MS method proved high selectivity and consistent analytical confidence. The limits of detection (LOD) ranged between 2 and 3 μg L-1. Intra-day and inter-day RSDs fluctuated between 1.7 - 13.4% and 0.4–10.9%, respectively. Weighted calibration revealed good linearity within the studied range (5–200 μg L−1) and guaranteed appropriate accuracy (relative residues < 25%). MSPE with GO-Fe3O4 demonstrated to be a practical, fast, efficient, high-throughput, and environmental-friendly sample preparation technique for determining of gingerols in commercial products, and its hyphenation with LC-MS/MS analysis yield a valuable analytical tool for the confident quality control of commercial ginger-containing products.

Efficient enrichment and analysis of atrazine and its degradation products in Chinese Yam using accelerated solvent extraction and pipette tip solid-phase extraction followed by UPLC–DAD

The Chinese Yam (Dioscorea opposita) is cultivated widely in China. However, there are potential safety risks associated with herbicide residues in Chinese Yam because of its characteristics and unregulated use of pesticides. We developed an efficient sample preparation technique for simultaneous analysis of atrazine and its degradation products in Chinese Yam using accelerated solvent extraction and pipette tip solid-phase extraction followed by UPLC-DAD analysis. Molecularly imprinted polymer (MIP) were used as the adsorbent material. Method performance was evaluated and our results showed the limit of quantification was 1–3 μg/ kg, with recoveries in the range 79.2–103.8%. When compared with other methods, this method provided superior performance in terms of sensitivity, accuracy, and precision. This method is, therefore, expected to be useful in supervised residue analysis.

СРАВНЕНИЕ ИСП-МС АНАЛИЗА ГЕОЛОГИЧЕСКИХ ОБРАЗЦОВ В ВАРИАНТЕ РАСТВОРОВ И ЛАЗЕРНОЙ АБЛЯЦИИ СТЕКОЛ

Актуальность исследования связана с необходимостью опробования нового эффективного способа пробоподготовки геологических образцов, подходящего для массового многоэлементного ИСП-МС анализа, снижающего общие временные затраты и обеспечивающего возможность определения широкого набора элементов в рамках одного измерительного цикла с применением лазерной абляции (ЛА-ИСП-МС). Цель исследования состоит в сравнении метрологических характеристик двух методик ИСП-МС анализа, основанных на разных способах пробоподготовки силикатных пород – переведении твердого образца в раствор после сплавления с метаборатом лития и использовании готовых боратных стекол для ЛА-ИСП-МС анализа. Объекты: международные и отечественные стандартные образцы состава естественных горных пород: базальты – BHVO-1, BHVO-2, BCR-2, граниты – СГ-1а, СГ-3, габбро СГД-1а, трапп СТ-1а. Методы: масс-спектрометрия с индуктивно-связанной плазмой (ИСП-МС), лазерная абляция с масс-спектрометрическим окончанием (ЛА-ИСП-МС). Результаты исследования показали, что разработанные методики многоэлементного ИСП-МС и ЛА-ИСП-МС анализа позволяют определять широкий набор элементов в геологических образцах в рамках одного измерительного цикла в высоком, среднем и низком разрешении при использовании для внешней градуировки стандартных образцов с аналогичной матрицей и внутреннего стандарта для учета и снижения матричного влияния и дрейфа приборных параметров. Правильность определения всех элементов в соответствии с разработанными методиками доказана сравнением результатов анализа шести стандартных геологических образцов с принятыми значениями. ЛА-ИСП-МС отличается экспрессностью за счет отсутствия стадии пробоподготовки, при этом пределы обнаружения и погрешность определения для ЛА-ИСП-МС методики выше, чем при ИСП-МС анализе растворов.

Fast quantitative urinary proteomic profiling workflow for biomarker discovery in kidney cancer

BackgroundUrine has evolved as a promising body fluids in clinical proteomics because it can be easily and noninvasively obtained and can reflect physiological and pathological status of the human body. Many efforts have been made to characterize more urinary proteins in recent years, but few have focused on the analysis throughput and detection reproducibility. Increasing the urine proteomic profiling throughput and reproducibility is urgently needed for discovering potential biomarker in large cohorts.MethodsIn this study, we developed a fast and robust workflow for streamlined urinary proteome analysis. The workflow integrate highly efficient sample preparation technique and urinary specific data-independent acquisition (DIA) approach. The performance of the workflow was systematically evaluated and the workflow was subsequently applied in a proof-of-concept urine proteome study of 21 kidney cancer (KC) patients and 22 healthy controls.ResultsWith this workflow, the entire sample preparation process takes less than 3 h and allows multiplexing on standard centrifuges. Without pre-fractionation, our newly developed DIA method allows quantitative analysis of ~ 1000 proteins within 80 min of MS time (~ 15 samples/day). The quantitation accuracy of the whole workflow was excellent with median CV of 9.1%. The preliminary study on KC identified 125 significantly changed proteins.ConclusionsThe result suggested the feasibility of applying the high throughput workflow in extensive urinary proteome profiling and clinical relevant biomarker discovery.

Implementation of antimicrobial peptides for sample preparation prior to nucleic acid amplification in point-of-care settings

ABSTRACTBackground: A variety of sample preparation techniques are used prior to nucleic acid amplification. However, their efficiency is not always sufficient and nucleic acid purification remains the preferred method for template preparation. Purification is difficult and costly to apply in point-of-care (POC) settings and there is a strong need for more robust, rapid, and efficient biological sample preparation techniques in molecular diagnostics.Methods: Here, the authors applied antimicrobial peptides (AMPs) for urine sample preparation prior to isothermal loop-mediated amplification (LAMP). AMPs bind to many microorganisms such as bacteria, fungi, protozoa and viruses causing disruption of their membrane integrity and facilitate nucleic acid release.Results: The authors show that incubation of E. coli with antimicrobial peptide cecropin P1 for 5 min had a significant effect on the availability of template DNA compared with untreated or even heat treated samples resulting in up to six times increase of the amplification efficiency.Conclusion: These results show that AMPs treatment is a very efficient sample preparation technique that is suitable for application prior to nucleic acid amplification directly within biological samples. Furthermore, the entire process of AMPs treatment was performed at room temperature for 5 min thereby making it a good candidate for use in POC applications.

Application of ionic liquid-based ultrasonic-assisted extraction followed by rapid resolution LC-ESI/MS to chromone analysis from radix saposhnikoviae

Ionic liquid-based ultrasonic-assisted extraction (ILUAE) was successfully applied to the extraction of the four chromones (prim-O-glucosylcimifugin, cimifugin, 5-O-methylvisammioside, and sec-O-glucosylhamaudol) from Saposhnikovia divaricata (Radix Saposhnikoviae) for the first time. A series of l-alkyl-3-methylimidazolium ILs differing in anion and cation compositions was evaluated for extraction efficiency, and [C3MIM]Br was selected as the optimal solvent. In addition, ultrasound extraction parameters were optimized, and the chromones were directly quantified and analyzed by rapid resolution liquid chromatography-electrospray ionization/mass spectrometry (RRLC-ESI/MS). The optimal conditions were as follows: 0.4 M concentration of [C3MIM]Br, 20:1 solvent to solid ratio, and ultrasonic time, temperature, and frequency of 5 min, 40 °C, and 50 kHz, respectively. This approach obtained the highest extraction yield of 10.188 ± 0.473 mg g−1 for total chromones. Compared with regular UAE, the proposed approach exhibited a higher efficiency (61.56% increase) and shorter extraction time (nine times shorter). Also, ILUAE was an efficient, rapid, and simple sample preparation technique for extraction of chromones, and the established RRLC-DAD method could serve as a rapid and effective technique for extracting chromones from Radix Saposhnikoviae.

Deep Metaproteomics Approach for the Study of Human Microbiomes.

Host-microbiome interactions have been shown to play important roles in human health and diseases. Most of the current studies of the microbiome have been performed by genomic approaches through next-generation sequencing. Technologies, such as metaproteomics, for functional analysis of the microbiome are needed to better understand the intricate host-microbiome interactions. However, significant efforts to improve the depth and resolution of gut metaproteomics are still required. In this study, we combined an efficient sample preparation technique, high resolution mass spectrometry, and metaproteomic bioinformatics tools to perform ultradeep metaproteomic analysis of human gut microbiome from stool. We reported the deepest analysis of the microbiome to date with an average of 20 558 protein groups identified per sample analysis. Moreover, strain resolution taxonomic and pathway analysis using deep metaproteomics revealed strain level variations, in particular for Faecalibacterium prausnitzii, in the microbiome from the different individuals. We also reported that the human proteins identified in stool samples are functionally enriched in extracellular region pathways and in particular those proteins involved in defense response against microbial organisms. Deep metaproteomics is a promising approach to perform in-depth microbiome analysis and simultaneously reveals both human and microbial changes that are not readily apparent using the standard genomic approaches.

Electrokinetic device design and constraints for use in high conductance solutions.

The quest for new cell-free DNA and exosome biomarker-based molecular diagnostics require fast and efficient sample preparation techniques. Conventional methods for isolating these biomarkers from blood are both time-consuming and laborious. New electrokinetic microarray devices using dielectrophoresis (DEP) to isolate cell-free DNA and exosome biomarkers have now greatly improved the sample preparation process. Nevertheless, these devices still have some limitations when used with high conductance biological fluids, e.g. blood, plasma, and serum. This study demonstrates that electrochemical damage may occur on the platinum electrodes of DEP microarray devices. It further examines two model device designs that include a parallel wire arrangement and a planar array. Effective isolation of fluorescent beads with parallel wires is shown under low-conductance conditions (10-4 S/m), but electrothermal flow overcomes DEP forces under high conductance conditions (>0.1S/m). Planar devices are shown to be effective under high conductance conditions (∼1S/m) without the deleterious effects of electrothermal flow. This study provides new insights into design compromises and limitations for producing future electrokinetic devices for better performance with high conductance solutions.

Recent developments on field gas extraction and sample preparation methods for radiokrypton dating of groundwater

Summary Current and foreseen population growths will lead to an increased demand in freshwater, large quantities of which is stored as groundwater. The ventilation age is crucial to the assessment of groundwater resources, complementing the hydrological model approach based on hydrogeological parameters. Ultra-trace radioactive isotopes of Kr ( 81 Kr and 85 Kr) possess the ideal physical and chemical properties for groundwater dating. The recent advent of atom trap trace analyses (ATTA) has enabled determination of ultra-trace noble gas radioisotope abundances using 5–10 μ L of pure Kr. Anticipated developments will enable ATTA to analyze radiokrypton isotope abundances at high sample throughput, which necessitates simple and efficient sample preparation techniques that are adaptable to various sample chemistries. Recent developments of field gas extraction devices and simple and rapid Kr separation method at the University of Chicago are presented herein. Two field gas extraction devices optimized for different sampling conditions were recently designed and constructed, aiming at operational simplicity and portability. A newly developed Kr purification system enriches Kr by flowing a sample gas through a moderately cooled (138 K) activated charcoal column, followed by a gentle fractionating desorption. This simple process uses a single adsorbent and separates 99% of the bulk atmospheric gases from Kr without significant loss. The subsequent two stages of gas chromatographic separation and a hot Ti sponge getter further purify the Kr-enriched gas. Abundant CH4 necessitates multiple passages through one of the gas chromatographic separation columns. The presented Kr separation system has a demonstrated capability of extracting Kr with > 90% yield and 99% purity within 75 min from 1.2 to 26.8 L STP of atmospheric air with various concentrations of CH4. The apparatuses have successfully been deployed for sampling in the field and purification of groundwater samples.

Mixed hemimicelles solid-phase extraction based on sodium dodecyl sulfate-coated nano-magnets for selective adsorption and enrichment of illegal cationic dyes in food matrices prior to high-performance liquid chromatography-diode array detection detection

In this study, mixed hemimicelles solid-phase extraction (MHSPE) based on sodium dodecyl sulfate (SDS) coated nano-magnets Fe3O4 was investigated as a novel method for the extraction and separation of four banned cationic dyes, Auramine O, Rhodamine B, Basic orange 21 and Basic orange 22, in condiments prior to HPLC detection. The main factors affecting the extraction of analysts, such as pH, surfactant and adsorbent concentrations and zeta potential were studied and optimized. Under optimized conditions, the proposed method was successful applied for the analysis of banned cationic dyes in food samples such as chili sauce, soybean paste and tomato sauce. Validation data showed the good recoveries in the range of 70.1–104.5%, with relative standard deviations less than 15%. The method limits of determination/quantification were in the range of 0.2–0.9 and 0.7–3μgkg−1, respectively. The selective adsorption and enrichment of cationic dyes were achieved by the synergistic effects of hydrophobic interactions and electrostatic attraction between mixed hemimicelles and the cationic dyes, which also resulted in the removal of natural pigments interferences from sample extracts. When applied to real samples, RB was detected in several positive samples (chili powders) within the range from 0.042 to 0.177mgkg−1. These results indicate that magnetic MHSPE is an efficient and selective sample preparation technique for the extraction of banned cationic dyes in a complex matrix.

Low density solvent based dispersive liquid-liquid microextraction and preconcentration of multiresidue pesticides in environmental waters for liquid chromatographic analysis

A simple, efficient and selective sample preparation technique using low density based dispersive liquid-liquid microextraction has been developed for the analysis of nine multiresidue pesticides inculding six sulfonylurea and three organophosphorus pesticides in environmental waters by HPLC-diod array detector. Various experimental parameters affecting the extraction efficiency were investigated. Under the optimum experimental conditions, matrix-matched calibration curves were established in groundwater and good linearities were obtained with coefficient of determination (r2) of 0.990 or better. The limits of detection and quantification were in the ranges 0.8–3.3 and 2.5–11.0 µg/L, at a signal-to-noise ratio of 3 and 10, respectively. The relative standard deviations of the precision studies were varied over the range of 0.2–13%. The proposed method was successfully applied to selective extraction of the target pesticide residues in different environmental waters and acceptable recoveries, in the range 81–121%, were obtained.

Vacuum-powered bubble-assisted solvent extraction followed by macroporous resin enrichment for isolation of podophyllotoxin from Sinopodophyllum emodi.

A vacuum-powered bubble-assisted solvent extraction (VBE) technique was used to extract podophyllotoxin from the root of Sinopodophyllum emodi. We optimized the VBE procedure and showed it had the highest efficiency of extraction compared to other conventional extraction techniques. Based upon the results of single-factor experiments, a three-factor, three-level experiment design was developed by application of a Box-Behnken design. The method was validated by stability, repeatability and recovery experiments. The optimal conditions were: solvent, 60% (v/v) ethanol; particle size of the sample, 60-80 mesh; soak time, 2h; liquid/solid ratio, 21L/kg; air flow, 32mL/min; vacuum-powered bubble extraction time, 65min. The VBE method we developed achieved efficient extraction of podophyllotoxin from S. emodi. The podophyllotoxin extracted can be enriched and separated by an HPD300 macroporous resin adsorption and desorption process. The results indicated that VBE is a convenient, rapid and efficient sample preparation technique.

Effects of Sample Processing Techniques and Instrument Model on Matrix Effect in LC-MS/MS for a Model Drug

The common perception that utilization of liquid chromatography-mass spectrometer (LC-MS) guarantees selectivity has now been challenged by a number of reported examples for the lack of selectivity due to ion suppression or enhancement caused by the sample matrix and interference from metabolites. It has been observed that an efficient sample preparation technique can reduce the matrix effect significantly. This research aims to evaluate the synergistic effect of sample processing technique and instrument model on matrix effect in LC-MS analysis by post-extraction addition and post-column infusion methods. Accordingly, different sample processing techniques, i.e., protein precipitation, liquid–liquid extraction, and solid phase extraction were evaluated for model drug, Esomeprazole, and samples processed with all the three techniques analyzed on three different MS models of AB Sciex, i.e., API-3000, API-3200, and API-4000. Samples were analyzed using Discovery C18 column and mobile phase acetonitrile: 10 mM ammonium acetate buffer (pH 4.8–5.2) :: 80:20, v/v. Results visibly showed that matrix effect was nullified by analyzing samples of liquid–liquid extraction technique on API-3200 and indicated that instrument model and sample processing technique have synergistic effect in contribution to matrix effect. Therefore, one should evaluate matrix effect in the early stage of method development and combine effective sample-processing technique with appropriate instrument model.

Influence of Ionization and Sample Processing Techniques on Matrix Effect of a Pulmonary Artery Antihypertensive Drug

The issue of selectivity due to ion suppression or enhancement caused by the sample matrix has become a challenge in mass spectrometric (MS) analysis. It has been pragmatic that an efficient sample preparation method can reduce the matrix effect significantly. The study was planned to assess the effect of ionization type and sample preparation techniques on the presence or absence of matrix effect in quantitative liquid chromatography-tandem mass spectrometric (LC-MS/MS) analysis of bosentan (BSN) by post extraction addition method. Different sample processing techniques, i.e., protein precipitation (PPT), liquid–liquid extraction (LLE), and solid phase extraction (SPE) were evaluated. The sample were analyzed by both LC-electrospray ionization (ESI)-MS/MS and LCatmospheric pressure chemical ionization (APCI)-MS/MS. Chromatographic partition achieved on an Aquasil C18 column (100 × 2.1 mm, five μm). The mobile phase consisted of ammonium formate (pH 4.5) and methanol (10:90 v/v) in non-gradient elution mode. Our results demonstrated that both ionization sources showed matrix effect, but ESI was more prone to matrix effect than APCI. Sample processing techniques could decrease or increase matrix effect. PPT was found to be least efficient sample preparation technique, due to the presence of many remaining matrix constituents and often resulting in significant matrix effects. LLE also provided clean final extracts. However, the efficiency of LLE was lower than SPE. SPE methods resulted in cleaner extracts and considerably reduced the levels of residual matrix components from plasma samples that ultimately leads to significant reduction in matrix effects. LC-APCI-MS/MS was found to be the ionization of choice for quantitative analysis of BSN and other drugs with similar physicochemical properties. The combination of SPE and the ionization source offer a major advantage in reducing matrix effects resulting from plasma components and in improving the selectivity and sensitivity of drug analysis.

Recent advances in the analysis of gibberellins plant hormones

Gibberellins (GAs) are a class of phytohormones that exert profound and diverse effects on plant growth and development, such as seed germination and leaf expansion. Up to now, 136 members of GAs have been identified and recognized. All known GAs are diterpenoid acids with similar chemical structures, only double bonds, hydroxyl numbers and locations on gibberellin alkane skeleton are different. However, the content of GAs in plants is of ultra trace levels (usually at ng/g and even pg/g levels) with little ultraviolet (UV) absorption, no fluorescence and no distinguishing chemical characteristics. Moreover, the matrix of plant samples is complicated. Thus, quantification of GAs is always extremely difficult. Nowadays, the bottle necks for the study of GAs in plants are due to the lack of efficient sample preparation and sensitive detection techniques. This article reviews the analytical methods for determination of GAs in recent years, hoping to provide some references to develop new methods and techniques.

Extraction and Chromatographic Determination of Shikimic Acid in Chinese Conifer Needles with 1-Benzyl-3-methylimidazolium Bromide Ionic Liquid Aqueous Solutions

An ionic liquids-based ultrasound-assisted extraction (ILUAE) method was successfully developed for extracting shikimic acid from conifer needles. Eleven 1-alkyl-3-methylimidazolium ionic liquids with different cations and anions were investigated and 1-benzyl-3-methylimidazolium bromide solution was selected as the solvent. The conditions for ILUAE, including the ionic liquid concentration, ultrasound power, ultrasound time, and liquid-solid ratio, were optimized. The proposed method had good recovery (99.37%–100.11%) and reproducibility (RSD, n = 6; 3.6%). ILUAE was an efficient, rapid, and simple sample preparation technique that showed high reproducibility. Based on the results, a number of plant species, namely, Picea koraiensis, Picea meyeri, Pinus elliottii, and Pinus banksiana, were identified as among the best resources of shikimic acid.

Development of sample preparation method for eleutheroside B and E analysis in Acanthopanax senticosus by ionic liquids-ultrasound based extraction and high-performance liquid chromatography detection

An ionic liquids-based ultrasonic-assisted extraction (ILUAE) method was successfully developed for extracting eleutheroside B and E from Radix Acanthopanax senticosus. Thirteen 1-alkyl-3-methylimidazolium ionic liquids with different cations and anions were investigated and 1-butyl-3-methylimidazolium bromide ([C4mim]Br) solution was selected as the solvent. The conditions for ILUAE, including the ionic liquid concentration, soaking time, ultrasonic power, ultrasonic time, solid–liquid ratio and number of extraction cycles, were optimized. With the proposed method, the energy consumption time was reduced to 30min, whereas conventional method requires about 4h. The proposed method had good recovery (97.96–103.39%) and reproducibility (RSD, n=5; 3.3% for eleutheroside B, 4.6% for eleutheroside E). ILUAE was an efficient, rapid and simple sample preparation technique that showed high reproducibility and was environmental friendly.