Evaluation of the extraction efficiency of thermally labile bioactive compounds in Gastrodia elata Blume by pressurized hot water extraction and microwave-assisted extraction

Our earlier work showed that the stability of the bioactive compounds gastrodin (GA) and vanillyl alcohol (VA) in Gastrodia elata Blume behaved differently with varying compositions of water-ethanol using pressurized liquid extraction (PLE) at room temperature. To have a better understanding of the extraction process of these thermally labile compounds under elevated temperature conditions, pressurized hot water extraction (PHWE) and microwave-assisted extraction (MAE) methods were proposed. PHWE and MAE showed that GA and VA could be extracted using pure water under optimized conditions of temperature and extraction time. The extraction efficiency of GA and VA by the proposed methods was found to be higher or comparable to heating under reflux using water. The marker compounds present in the plant extracts were determined by RP-HPLC. The optimized conditions were found to be different for the two proposed methods on extraction of GA and VA. The method precision (RSD, n=6) was found to vary from 0.92% to 3.36% for the two proposed methods on different days. Hence, PHWE and MAE methods were shown to be feasible alternatives for the extraction of thermally labile marker compounds present in medicinal plants.     

Evaluation of surfactant‐assisted pressurized hot water extraction for marker compounds in Radix Codonopsis pilosula using liquid chromatography and liquid chromatography/electrospray ionization mass spectrometry

In the move towards the elimination of organic solvents in the extraction process in botanicals, a new method combining surfactant and pressurized hot water extraction (PWHE) with an applied temperature below the boiling point and lower pressure from 10 to 20 bar was developed for the analysis of marker compounds that are reasonably hydrophobic such as tetradeca‐4E,12E‐diene‐8,10‐diyne‐1,6,7‐triol and tetradeca‐4E,12E‐diene‐8,10‐diyne‐1,6,7‐triol‐O‐β‐D‐glucoside in Radix Codonopsis pilosula (DangShen). Because reference substances for the proposed botanicals were not available, a method was developed to isolate the marker compounds in Radix Codonopsis pilosula. Other than surfactant‐assisted PHWE, the marker compounds present in Radix Codonopsis pilosula were extracted using pressurized liquid extraction (PLE) with methanol and PHWE with a mixture of water/ethanol (80:20). The extracts were analyzed using liquid chromatography and liquid chromatography/electrospray ionization mass spectrometry. With surfactant‐assisted PHWE, the effects of different added surfactants such as sodium dodecyl sulfate and Triton X‐100 was studied. Surfactant assisted PHWE with Triton X‐100 proved to be at least equivalent or better compared to Soxhlet extraction in terms of quantitative analysis of marker compounds in Radix Codonopsis pilosula. The method precision was less than 8% (RSD, n = 6). The presence of surfactants in PHWE was found to enhance the solubility of target compounds naturally occurring in medicinal plants.     

Pressurized hot water extraction of berberine, baicalein and glycyrrhizin in medicinal plants

Pressurized hot water extraction (PHWE) using a laboratory made system was applied for the extraction of thermally labile and reasonably polar components such as berberine in coptidis rhizoma, glycyrrhizin in radix glycyrrhizae/liquorice and baicalein in scutellariae radix. PHWE was carried out dynamically at a flow of 1 ml/min, temperature between 95 and 140 °C, an applied pressure of 10-20 bar and extraction time of 40 min. Extraction by PHWE was found to give efficiencies comparable to Soxhlet extraction for baicalein in scutellariae radix and sonication for berberine in coptidis rhizoma, and glycyrrhizin in radix glycyrrhizae. Effects of ethanol added into the water used in PHWE were explored. Pressurized liquid extraction (PLE) with methanol as solvent was used for extraction of baicalein in scutellariae radix. The marker compounds present in the various medicinal plant extracts were determined by gradient elution HPLC.     

Evaluation of pressurized liquid extraction and pressurized hot water extraction for tanshinone I and IIA in Salvia miltiorrhiza using LC and LC-ESI-MS

Pressurized liquid extraction (PLE) and pressurized hot water extraction (PHWE) using a laboratory-made system are applied for the extraction of thermally labile components such as tanshinone I and IIA in Salvia miltiorrhiza. PLE and PHWE are carried out dynamically at a flow of 1 mL/min, temperature between 95-140 degrees C, applied pressure of 10-20 bars, and extraction times of 20 and 40 min, respectively. Effects of ethanol added into the water used in PHWE are explored. PLE is found to give comparable or higher extraction efficiencies compared with PHWE with reference to Soxhlet extraction for tanshinone I and IIA in Salvia miltiorrhiza. The tanshinone I and IIA present in the various medicinal plant extracts are determined by liquid chromatography and liquid chromatography-mass spectrometry.     

Validation of green‐solvent extraction combined with chromatographic chemical fingerprint to evaluate quality of Stevia rebaudiana Bertoni

An approach that combined green‐solvent methods of extraction with chromatographic chemical fingerprint and pattern recognition tools such as principal component analysis (PCA) was used to evaluate the quality of medicinal plants. Pressurized hot water extraction (PHWE) and microwave‐assisted extraction (MAE) were used and their extraction efficiencies to extract two bioactive compounds, namely stevioside (SV) and rebaudioside A (RA), from Stevia rebaudiana Bertoni (SB) under different cultivation conditions were compared. The proposed methods showed that SV and RA could be extracted from SB using pure water under optimized conditions. The extraction efficiency of the methods was observed to be higher or comparable to heating under reflux with water. The method precision (RSD, n = 6) was found to vary from 1.91 to 2.86% for the two different methods on different days. Compared to PHWE, MAE has higher extraction efficiency with shorter extraction time. MAE was also found to extract more chemical constituents and provide distinctive chemical fingerprints for quality control purposes. Thus, a combination of MAE with chromatographic chemical fingerprints and PCA provided a simple and rapid approach for the comparison and classification of medicinal plants from different growth conditions. Hence, the current work highlighted the importance of extraction method in chemical fingerprinting for the classification of medicinal plants from different cultivation conditions with the aid of pattern recognition tools used.     

Pressurized hot water extraction (PHWE)

Pressurized hot water extraction (PHWE) has become a popular green extraction method for different classes of compounds present in numerous kinds of matrices such as environmental, food and botanical samples. PHWE is also used in sample preparation to extract organic contaminants from foodstuff for food safety analysis and soils/sediments for environmental monitoring purposes. The main parameters which influence its extraction efficiency are namely the temperature, extraction time, flow rates and addition of modifiers/additives. Among these different parameters studied, temperature is described as the most important one. It is reported that the extraction of certain compounds is rather dependent on pressurized water with different applied temperature. Thus, the stability and reduced solubilities of certain compounds at elevated temperatures are highlighted in this review. With some modifications, a scaled-up PHWE could extract a higher amount of desirable compounds from solid and powdered samples such as plant and food materials. The PHWE extracts from plants are rich in chemical compounds or metabolites which can be a potential lead for drug discovery or development of disease-resistant food crops.     

Pressurized hot water extraction of insecticides from process dust--comparison with supercritical fluid extraction

Pressurized hot liquid water and steam were used to investigate the possibilities of extracting insecticides (carbofuran, carbosulfan, and imidacloprid) from contaminated process dust remaining from seed-pellet production. Extraction temperature was the most important parameter in influencing the extraction efficiency and rate of extraction, while varying the pressure had no profound effect. A clean-up procedure of the water extracts using solid phase extraction (SPE) was found to be necessary prior to final analysis by high-performance liquid chromatography (HPLC). Quantitative extraction (compared to a validated organic solvent extraction method) of imidacloprid was obtained at temperatures of 100-150 degrees C within 30 min extraction time. Temperatures above 150 degrees C were required to extract carbofuran efficiently. The most non-polar analyte of the investigated compounds, carbosulfan, gave no detectable concentrations with pressurized hot water extraction (PHWE). One reason might be its low solubility in water, and when attempts are made to increase its solubility by increasing the temperature it may degrade to carbofuran. This can explain recovery values above 100% for carbofuran at higher temperatures. A comparison of the PHWE results and those obtained with supercritical fluid extraction (SFE) revealed that PHWE is advantageous for polar compounds, where the solubility of the analyte in water is high enough that lower temperatures can be used. For non-polar compounds carbon dioxide based extraction is preferred unless the target analyte is highly thermostable.     

Determination of pyrrolizidine alkaloids in comfrey by liquid chromatography-electrospray ionization mass spectrometry

Symphytum officinale L. (comfrey) is a medicinal plant commonly used in decoctions and aliments. Besides therapeutic bioactive compounds present in the herb, it is found to contain hepatotoxic pyrrolizidine alkaloids (PAs), such as lycopsamine and others. In the present study, PAs such as lycopsamine, echimidine and lasiocarpine were determined using electrospray liquid chromatography-mass spectrometry (LC-MS) with the method precision (relative standard deviation, RSD) <10%. Detection of lycopsamine, symviridine and their N-oxides could be confirmed with a newly developed method based on HPLC ion-trap and orbitrap MS with electrospray ionization interface. With LC-MS, quantitative analysis of lycopsamine in the botanical extract was carried out. The effect of extraction solvent was optimized by sonication and methanol: H₂O (50:50) was selected. Then a rapid method based on pressurized hot water extraction (PHWE) was employed for the extraction of lycopsamine from comfrey followed by the comparison with heating under reflux with the RSD ranging from 2.49% to 19.32%. Our results showed a higher extraction efficiency for heating under reflux compared with PHWE. It was proposed that the lower extraction efficiency for PHWE was attributable to dissolved nitrogen from air which caused the reduction in the solubility of lycopsamine in the compressed hot solvent. In this study, quantitative analysis of PAs in comfrey was demonstrated. In addition, it was found that the use of subcritical water for extractions depended on the physical properties of the dissolved solutes and their tendency to degrade under the chosen extraction conditions.     

Optimization of Extraction of Bioactive Compounds from Baphicacanthus cusia Leaves by Hydrophobic Deep Eutectic Solvents

Deep eutectic solvents (DESs) are considered as efficient and green solvents for the extraction of bioactive compounds from medicinal plants. In this work, a novel method of DES-based ultrasound-assisted extraction of bioactive compounds from Baphicacanthus cusia leaves (BCL) was established. Systematic screening and the morphology of the original and treated BCL were observed with scanning electron microscopy to determine the extraction efficiency of different solvents. The extraction conditions were optimized by Box–Behnken design (BBD) tests and the optimal extraction conditions were as follows: lactic acid/L-menthol ratio of 5: 2 (mol/mol), solid–liquid ratio of 80.0 mL/g and temperature of 60.5 °C. The extraction yields of tryptanthrin, indigo and indirubin reached 0.356, 1.744 and 0.562 mg/g, respectively. The results of a 2,2-diphenyl-1-picrylhydrazy (DPPH) radical scavenging activity test indicated the feasibility of DESs in the extraction of bioactive compounds. This study indicated that L-menthol/lactic acid was a green and efficient solvent for the extraction of bioactive compounds from BCL, and DES-based ultrasound-assisted extraction could be used as an effective application strategy for the extraction of bioactive compounds from medicinal plants.     

Study of polyethylene glycol as a green solvent in the microwave-assisted extraction of flavone and coumarin compounds from medicinal plants

In this paper, the application of polyethylene glycol (PEG) aqueous solution as a green solvent in microwave-assisted extraction (MAE) was firstly developed for the extraction of flavone and coumarin compounds from medicinal plants. The PEG solutions were optimized by a mono-factor test, and the other conditions of MAE including the size of sample, liquid/solid ratio, extraction temperature and extraction time were optimized by means of an orthogonal design L(9) (3(4)). Subsequently, PEG-MAE, organic solvent-MAE, and conventional heating reflux extraction (HRE) were evaluated with nevadensin extraction from Lysionotus pauciflorus, aesculin and aesculetin extraction from Cortex fraxini. Furthermore, the mechanism of PEG-MAE was investigated, including microwave-absorptive property and viscosity of PEG solutions, the kinetic mechanism of PEG-MAE and different microstructures of those samples before and after extraction. Under optimized conditions, the extraction yields of nevadensin from L. pauciflorus, aesculin and aesculetin from C. fraxini were 98.7%, 97.7% and 95.9% in a one-step extraction, respectively. The recoveries of nevadensin, aesculin and aesculetin were in the range of 92.0-103% with relative standard derivation lower than 3.6% by the proposed procedure. Compared with organic solvent-MAE and conventional extraction procedures, the proposed methods were effective and alternative for the extraction of flavone and coumarin compounds from medicinal plants. On the basis of the results, PEG solution as a green solvent in the MAE of active compounds from medicinal plants showed a great promising prospect.     

Pressurized liquid extraction in environmental analysis

A critical evaluation of recent literature utilizing pressurized liquid extraction (PLE) for environmental analysis is presented by compound class. Overall, the extraction efficiency of PLE, using the appropriate solvent, temperature and pressure for extraction, is similar to that of Soxhlet extraction. PLE has been used for some classes of compounds that are thermally labile (e.g., explosives) and may require acidic conditions for extraction (e.g., organometallic compounds). References to recent applications are presented emphasizing studies which utilize unspiked, natural matrices and studies that compare PLE to alternate extraction techniques.     

Extending the applicability of pressurized hot water extraction to compounds exhibiting limited water solubility by pH control: curcumin from the turmeric rhizome

Pressurized hot water extraction (PHWE, also known as subcritical water extraction) is commonly considered to be an environmentally friendly extraction technique that could potentially replace traditional methods that use organic solvents. Unfortunately, the applicability of this technique is often limited by the very low water solubility of the target compounds, even at high temperatures. In this paper, the scope for broadening the applicability of PHWE by adjusting the pH of the water used in the extraction is demonstrated in the extraction of curcumin (which exhibits very limited water solubility) from untreated turmeric (Curcuma longa L.) rhizomes. Although poor extraction yields were obtained, even at high temperatures when using degassed water or neutral phosphate buffer as the extraction medium, yields exceeding those obtained by Soxhlet extraction were achieved using highly acidic pH buffers due to curcumin protonation. The influence of the temperature, pH, and buffer concentration on the extraction yield were investigated in detail by means of a series of designed experiments. Optimized conditions for the extraction of curcumin from turmeric by PHWE were estimated at 197 °C using 62 g/L buffer concentration at pH 1.6. The relationships between these variables were subjected to statistical analysis using response surface methodology.     

Evaluation of surfactant assisted pressurized liquid extraction for the determination of glycyrrhizin and ephedrine in medicinal plants

Surfactant assisted pressurized liquid extraction (PLE) with a laboratory made system was applied for the extraction of glycyrrhizin in Radix glycyrrhizae/liquorice and ephedrine in Ephedra sinica. The proposed system set-up for this current work was simpler as no heating and back pressure regulator was required. Extraction with surfactant assisted PLE was carried out dynamically at a flow of 1.5 mL min⁻¹, at room temperature, under an applied pressure of 10-20 bar with an extraction time of 45-50 min. The extraction efficiencies of the proposed method using surfactants such as sodium dodecyl sulfate (SDS) and Triton X-100 were compared with sonication using organic solvent for different batches of medicinal plants materials. For the determination of glycyrrhizin in R. glycyrrhizae, the extraction efficiencies of surfactant assisted PLE with SDS and Triton X-100 was observed to be comparable with sonication. The method precision was found to vary from 1.6 to 2.6% (R.S.D., n = 6) on different days. For ephedrine in E. sinica, surfactant assisted PLE with SDS was found to give higher extraction efficiencies compared to Triton X-100. The overall method precision for surfactant assisted PLE with SDS for ephedrine in E. sinica was found to vary from 1.5 to 4.1% (R.S.D., n = 6) on different days. The marker compounds present in the various medicinal plant extracts were determined by gradient elution HPLC. Our data showed the possibility of PLE at room temperature and the advantages of eliminating the use of organic solvents in the extraction process.     

Pressurised hot water extraction in continuous flow mode for thermolabile compounds: extraction of polyphenols in red onions

Extraction and analysis of labile compounds in complex sample matrices, such as plants, is often a big analytical challenge. In this work, the use of a “green and clean” pressurised hot water extraction (PHWE) approach performed in continuous flow mode is explored. Experimental data for extraction and degradation kinetics of selected compounds were utilised to develop a continuous flow extraction (CFE) method targeting thermolabile polyphenols in red onions, with detection by high-performance liquid chromatography (HPLC)–diode array detection (DAD)–mass spectrometry (MS). Water containing ethanol and formic acid was used as extraction solvent. Method performance was focused on extraction yield with minimal analyte degradation. By adjusting the flow rate of the extraction solvent, degradation effects were minimised, and complete extraction could be achieved within 60 min. The CFE extraction yields of the polyphenols investigated were 80–90 % of the theoretically calculated quantitative yields and were significantly higher than the yields obtained by conventional methanol extraction and static batch extraction (70–79 and 58–67 % of the theoretical yields, respectively). The precision of the developed method was lower than 8 % expressed as relative standard deviation.

Determination of senkirkine and senecionine in Tussilago farfara using microwave-assisted extraction and pressurized hot water extraction with liquid chromatography tandem mass spectrometry

Tussilago farfara (Kuan Donghua) is an important Chinese herbal medicine which has been shown to contain many bioactive compounds and widely used to relieve cough and resolve phlegm. However, besides therapeutic bioactive compounds, this herb has been found to contain toxic pyrrolizidine alkaloids (PAs), mainly senkirkine and traces of senecionine. In this report, conditions for microwave-assisted extraction (MAE) and pressurized hot water extraction (PHWE) were optimized for the extraction of the PAs. The results were compared against heating under reflux. It was found that the binary mixture of MeOH:H₂O (1:1) acidified using HCl to pH 2-3 was the optimal solvent for the extraction of the PAs in the plant materials. Liquid chromatography (LC) with ultra-violet (UV) detection and electrospray ionization mass spectrometry (ESI-MS) in the positive mode was used for the determination and quantitation of senkirkine and senecionine in the botanical extract. The proposed extraction methods with LC/MS allow for the rapid detection of the major and the minor alkaloids in T. farfara in the presence of co-eluting peaks. With LC/MS, the quantitative analysis of PAs in the extract was done using internal standard calibration and the precision was found to vary from 0.6% to 5.4% on different days. The limits of detection (LODs) and limits of quantitation (LOQs) for MAE and PHWE were found to vary from 0.26 μg/g to 1.04 μg/g and 1.32 μg/g to 5.29 μg/g, respectively. The method precision of MAE and PHWE were found to vary from 3.7% to 10.4% on different days. The results showed that major and minor alkaloids extracted using MAE and PHWE were comparable to that by heating under reflux. Our data also showed that significant ion suppression was not observed in the analysis of senkirkine and senecionine in the botanical extracts with co-eluting peaks.     

Application of surfactants and microemulsions to the extraction of pyrene and phenanthrene from soil with three different extraction methods

In the present work, the use of surfactants and oil-in-water (O/W) microemulsions as alternative extractants in accelerated solvent extraction (ASE) for the extraction of polycyclic aromatic hydrocarbons (pyrene and phenanthrene) from soils was investigated. In particular, the effect of each individual component within the microemulsions, i.e., oil phase, surfactant and co-surfactatnt, and extraction conditions on the percentage recovery was systematically studied. When compared to the water and organic solvent, the important findings were that the common surfactant solutions at the concentrations above their critical micelle concentrations (CMC) were shown to enhance the percentage recovery at the lower extraction temperature. Moreover, the highest percentage recovery can be obtained using microemulsion as the extractant. The chemical component within the microemulsions and relative amounts of the oil phase appeared to play a much more significant role in ensuring high percentage recovery. Finally, an overall comparison between the percentage recoveries obtained with ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) and ASE using organic solvents, surfactants and microemulsions as extractants was exhibited.     

Optimal Template Removal from Molecularly Imprinted Polymers by Pressurized Hot Water Extraction

An optimal extraction method for the removal of templates from molecularly imprinted polymers (MIPs) is presented. The extraction method is based on pressurized hot water extraction (PHWE). PHWE was evaluated by application to three distinctly colored MIPs for chlorophyll (green), quercetin (yellow) and phthalocynine (dark blue) with subsequent monitoring of template removal and template bleeding by an ultraviolet spectrophotometer. The templates were washed-off and the extraction efficiency (EE) was compared to that of soxhlet and ultrasonic extraction methods. PHWE employed hot water at an optimal temperature of 220 °C, pressure of 50 bars and flow rate of 2 mL min^?1 to thoroughly wash-off the respective templates from their MIPs. The EE evaluated for PHWE was over 99.6% for all the MIPs with no subsequent or minimal template bleeding (<0.01%). The washing procedure was simple and relatively fast as it was achieved in 70 min at the most. At 95% confidence level (n = 3), soxhlet and ultrasonic recorded EE that was not significantly different (<94.5% in all cases) from that of PHWE (>99.6% in all cases). Soxhlet and ultrasonic had washing procedures that were slower (over 18 h) and employed large quantities (400 mL) of organic solvents modified with acids. The percentage relative standard deviations (%RSD) for the EE and recovery results were less than 2.3% in all cases indicating the high reproducibility of the method. Overall, the three methods performed comparably in extracting templates. PHWE seems to be the method of choice as it employed water which poses no environmental threat.     

Comparison of extraction techniques for extraction of bioactive molecules from Hypericum perforatum L. plant

Three methods commonly used for the extraction of bioactive molecules from natural plant material are compared. Dried Hypericum perforatum L. plant material is subjected to Soxhlet extraction, extraction by ultrasonication, and accelerated solvent extraction. The percentage of two bioactive compounds, hyperforin and hypericin, in the extracts is used as a parameter for comparison of the extraction procedure.     

Quantitative studies of rhubarb using quantitative analysis of multicomponents by single marker and response surface methodology

In this work, we developed a novel approach to evaluate the contents of bioactive components in rhubarb. The present method was based on the quantitative analysis of multicomponents by a single‐marker and response surface methodology approaches. The quantitative analysis of multicomponents by a single‐marker method based on high‐performance liquid chromatography coupled with photodiode array detection was developed and applied to determine the contents of 12 bioactive components in rhubarb. No significant differences were found in the results from the quantitative analysis of multicomponents by a single‐marker and the external standard method. In order to maximize the extraction of 12 bioactive compounds in rhubarb, the ultrasonic‐assisted extraction conditions were obtained by the response surface methodology coupled with Box–Behnken design. According to the obtained results, we showed that the optimal conditions would be as follows: proportion of ethanol/water 74.39%, solvent‐to‐solid ratio 24.07:1 v/w, extraction time 51.13 min, and extraction temperature 63.61°C. The analytical scheme established in this research should be a reliable, convenient, and appropriate method for quantitative determination of bioactive compounds in rhubarb.     

Factors Controlling Leaching of Polycyclic Aromatic Hydrocarbons from Petroleum Source Rock Using Nonionic Surfactant

The extraction of polycyclic aromatic hydrocarbons (PAHs) from petroleum source rock by nonionic surfactants with the assistance of microwave irradiation was investigated and the conditions for maximum yield were determined. The results showed that the extraction temperatures and type of surfactant have significant effects on extraction yields of PAHs. Factors such as surfactant concentration, irradiation power, sample/solvent ratio and mixing surfactants (i.e., mixture of surfactant at specific ratio) also influence the extraction efficiencies for these compounds. The optimum temperature for microwave-assisted nonionic surfactant extraction of PAHs from petroleum source rock was 120 °C and the best suited surfactant was Brij 35. The new method showed extraction efficiencies comparable to those afforded by the Soxhlet extraction method, but a reduction of the extraction times and environmentally friendliness of the new nonionic surfactant extraction system are clear advantages. The results also show that microwave-assisted nonionic surfactant extraction is a good and efficient green analytical preparatory technique for geochemical evaluation of petroleum source rock.