Development and validation of a green and sustainable procedure for the preparation of Perilla frutescens extracts.

A procedure combining supercritical CO2 and ultrasound-assisted (USC-CO2) extraction was developed to obtain rosmarinic acid (RA)-rich extracts from Perilla frutescens. Based on extraction yields and efficiencies, USC-CO2 was considered the best extraction method among the methods studied for obtaining RA from P. frutescens. The constant extraction rate period and the falling extraction rate period for USC-CO2 extraction of P. frutescens were 45 and 96 min long, respectively, and they were significantly shorter than those of traditional SC-CO2 (TSC-CO2) extraction. Furthermore, mass transfer coefficients were derived using the Sovová model for the fluid and solid phases from USC-CO2 extraction, with values of 9.752 × 10-3 and 4.203 × 10-3 min-1, respectively, which were obviously higher than those for TSC-CO2 extraction. Consequently, the theoretical solubilities of RA in the supercritical solvents used in dynamic USC-CO2 and TSC-CO2 extractions were estimated and found to be well correlated using three density-based models.

A green approach for the extraction and characterization of oridonin and ursolic and oleanolic acids from Rabdosia rubescens and its kinetic behavior.

An ultrasound-assisted supercritical carbon dioxide (USC-CO2) procedure was developed for the extraction of ursolic acid, oleanolic acid and oridonin from Rabdosia rubescens, with yields that were 9.84-10.46 and 15.43-21.10% higher than those of the conventional SC-CO2 and heat-reflux extractions, respectively. USC-CO2 uses a shorter extraction time (1.83-2.09 times) and less organic solvent (3.39-173.25 times) to operate at a lower extraction temperature (5-16 °C). The dominant component in the extract was oridonin, which may indicate that the kinetic behavior in the extraction system is predominated by that of oridonin. Furthermore, the USC-CO2 and conventional SC-CO2 dynamic extraction kinetics of oridonin from R. rubescens were well described by the second-order rate and Fick's second law models. The extraction rate constant, energy of activation for diffusion, Biot number and thermodynamic parameters were deduced from the data obtained. These results provide valuable insights into the USC-CO2 and conventional SC-CO2 procedures.

Development and characterization of a green procedure for apigenin extraction from Scutellaria barbata D. Don.

This study compared the use of ultrasound-assisted supercritical CO2 (USC-CO2) extraction to obtain apigenin-rich extracts from Scutellaria barbata D. Don with that of conventional supercritical CO2 (SC-CO2) extraction and heat-reflux extraction (HRE), conducted in parallel. This green procedure yielded 20.1% and 31.6% more apigenin than conventional SC-CO2 extraction and HRE, respectively. Moreover, the extraction time required by the USC-CO2 procedure, which used milder conditions, was approximately 1.9 times and 2.4 times shorter than that required by conventional SC-CO2 extraction and HRE, respectively. Furthermore, the theoretical solubility of apigenin in the supercritical fluid system was obtained from the USC-CO2 dynamic extraction curves and was in good agreement with the calculated values for the three empirical density-based models. The second-order kinetics model was further applied to evaluate the kinetics of USC-CO2 extraction. The results demonstrated that the selected model allowed the evaluation of the extraction rate and extent of USC-CO2 extraction.

Hepatoprotective effects of raspberry (Rubus coreanus Miq.) seed oil and its major constituents.

Raspberry seed is a massive byproduct of raspberry juice and wine but usually discarded. The present study employed a microwave-assisted method for extraction of raspberry seed oil (RSO). The results revealed that omega-6 fatty acids (linoleic acid and γ-linolenic acid) were the major constituents in RSO. Cellular antioxidant enzyme activity such as superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT) were investigated in HepG2 cells treated with RSO. Induction of the synthesis of several antioxidants in H2O2-exposed HepG2 cells was found. RSO increased the enzyme activity of SOD, CAT, and GPx in H2O2-exposed HepG2. Furthermore, RSO inhibited the phosphorylation of upstream mitogen-activated protein kinases (MAPK) such as c-Jun N-terminal kinase (c-JNK) and extracellular signal-regulated kinase (ERK). Taken together, the possible mechanisms to increase antioxidant enzyme activities in HepG2 may through the suppression of ERK and JNK phosphorylation. Raspberry seed oil exhibited good effects on the activities of the intracellular antioxidant enzymes and seems to protect the liver from oxidative stress through the inhibition of MAPKs.

Production of oridonin-rich extracts from Rabdosia rubescens using hyphenated ultrasound-assisted supercritical carbon dioxide extraction.

BACKGROUND: Among active components in Rabdosia rubescens, oridonin has been considered a key component and the most valuable compound because it has a wide range of activities beneficial to human health. To produce a high-quality oridonin extract, an alternative hyphenated procedure involving an ultrasound-assisted and supercritical carbon dioxide (HSC-CO2 ) extraction method to extract oridonin from R. rubescens was developed in this study. Fictitious solubilities of oridonin in supercritical CO2 (SC-CO2 ) with ultrasound assistance were measured by using the dynamic method at temperatures ranging from 305.15 K to 342.15 K over a pressure range of 11.5 to 33.5 MPa. RESULTS: Fictitious solubilities of oridonin at different temperatures and pressures were over the range of 2.13 × 10-6 to 10.09 × 10-6 (mole fraction) and correlated well with the density-based models, including the Bartle model, the Chrastil model, the Kumar and Johnston model and the Mendez-Santiago and Teja model, with overall average absolute relative deviations (AARDs) of 6.29%, 4.39%, 3.12% and 5.07%, respectively. CONCLUSION: Oridonin exhibits retrograde solubility behaviour in the supercritical state. Fictitious solubility data were further determined and obtained a good fit with four semi-empirical models. Simultaneously, the values of the total heat of solution, vaporisation and solvation of oridonin were estimated. © 2016 Society of Chemical Industry.

Extraction of α-humulene-enriched oil from clove using ultrasound-assisted supercritical carbon dioxide extraction and studies of its fictitious solubility.

Clove buds are used as a spice and food flavoring. In this study, clove oil and α-humulene was extracted from cloves using supercritical carbon dioxide extraction with and without ultrasound assistance (USC-CO2 and SC-CO2, respectively) at different temperatures (32-50°C) and pressures (9.0-25.0MPa). The results of these extractions were compared with those of heat reflux extraction and steam distillation methods conducted in parallel. The extracts obtained using these four techniques were analyzed using gas chromatography and gas chromatography/mass spectrometry (GC/MS). The results demonstrated that the USC-CO2 extraction procedure may extract clove oil and α-humulene from clove buds with better yields and shorter extraction times than conventional extraction techniques while utilizing less severe operating parameters. Furthermore, the experimental fictitious solubility data obtained using the dynamic method were well correlated with density-based models, including the Chrastil model, the Bartle model and the Kumar and Johnston model.

Determination of Oleanolic and Ursolic Acids in Hedyotis diffusa Using Hyphenated Ultrasound-Assisted Supercritical Carbon Dioxide Extraction and Chromatography.

Oleanolic acid (OA) and ursolic acid (UA) were extracted from Hedyotis diffusa using a hyphenated procedure of ultrasound-assisted and supercritical carbon dioxide (HSC-CO2) extraction at different temperatures, pressures, cosolvent percentages, and SC-CO2 flow rates. The results indicated that these parameters significantly affected the extraction yield. The maximal yields of OA (0.917 mg/g of dry plant) and UA (3.540 mg/g of dry plant) were obtained at a dynamic extraction time of 110 min, a static extraction time of 15 min, 28.2 MPa, and 56°C with a 12.5% (v/v) cosolvent (ethanol/water = 82/18, v/v) and SC-CO2 flowing at 2.3 mL/min (STP). The extracted yields were then analyzed by high performance liquid chromatography (HPLC) to quantify the OA and UA. The present findings revealed that H. diffusa is a potential source of OA and UA. In addition, using the hyphenated procedure for extraction is a promising and alternative process for recovering OA and UA from H. diffusa at high concentrations.

Ultrasound-assisted extraction and quantitation of oils from Syzygium aromaticum flower bud (clove) with supercritical carbon dioxide.

This study evaluated ultrasound-assisted supercritical carbon dioxide (USC-CO2) extraction for determining the extraction yields of oils and the contents of eugenol, ß-caryophyllene, eugenyl acetate and α-humulene from clove buds. Compared to traditional SC-CO2 extraction, USC-CO2 extraction might provide a 13.5% increase in the extraction yield for the oil while utilizing less severe operating parameters, such as temperature, pressure, CO2 flow rate and the time consumed by the process. Our results were comparable to those obtained using the heat reflux extraction method, though the yield was improved by 20.8% using USC-CO2. In kinetic studies, the USC-CO2 extraction of clove oil followed second-order kinetics. The activation energy for the oil extraction was 76.56kJ/mol. The USC-CO2 procedure facilitated the use of mild extraction conditions, improved extraction efficiency and the quality of products and is a potential method for industry.

Development of a hyphenated procedure of heat-reflux and ultrasound-assisted extraction followed by RP-HPLC separation for the determination of three flavonoids content in Scutellaria barbata D. Don.

A hyphenated procedure of heat-reflux and ultrasound-assisted extraction (HUAE), and an accurate high-performance liquid chromatographic (HPLC) method were developed for the determination of apigenin, baicalin and luteolin content in Scutellaria barbara D. Don. The suitable HUAE conditions for the extraction of target compounds from the herb were identified as an ultrasonic frequency of 40kHz, power of 185W, duty cycle of 75% (intermittent sonication), mean particle size of 0.355mm, extraction temperature of 50°C, ratio of solvent to raw material of 12:1 (mL/g), ethanol concentration of 60% (v/v), extraction time of 30min and three cycles. Compared with a traditional heat-reflux extraction method, the proposed method reduced the extraction time, extraction temperature and solvent consumption. Also, this HUAE method achieved superior apigenin, baicalin and luteolin yields. Furthermore, the developed HUAE-HPLC method was applied successfully for the simultaneous evaluation of three bioactive compounds in five samples of S. barbara D. Don obtained from different geographical regions. These results clearly demonstrated that the combined HUAE-HPLC process is feasible in the future commercialized manufacture of this highly valuable Chinese herbal medicine.

Development and validation of a modified ultrasound-assisted extraction method and a HPLC method for the quantitative determination of two triterpenic acids in Hedyotis diffusa.

In the present study, the oleanolic acid (OA) and ursolic acid (UA) contents ofHedyotis diffusa and H. corymbosa were determined by a rapid, selective and accurate method combining modified ultrasound-assisted extraction (MUAE) and HPLC. Compared with traditional extraction methods, MUAE reduced the extraction time, the extraction temperature and the solvent consumption and maximized the extraction yields of OA and UA. Furthermore, the combined MUAE-HPLC method was applied to quantitate OA and UA in plant samples and exhibited good repeatability, reproducibility and stability. The mean recovery studies (one extraction cycle) for OA and UA were between 91.3 and 91.7% with RSD values less than 4.5%. The pioneer method was further applied to quantitate OA and UA in six samples of H. diffusa and five samples of H. corymbosa. The results showed that the OA and UA content in the samples from different sources were significantly different. This report is valuable for the application of H. diffusa and H. corymbosa obtained from different regions in clinical research and pharmacology.

Optimisation of an ultrasound-assisted extraction followed by RP-HPLC separation for the simultaneous determination of oleanolic acid, ursolic acid and oridonin content in Rabdosia rubescens.

INTRODUCTION: Rabdosia rubescens is a commonly used herb in traditional Chinese medicine and contains diterpenoids, triterpenoids and various other compounds. Among these components, oridonin, oleanolic acid and ursolic acid have gained considerable interest concerning anti-cancer activities. However, there is no suitable currently available method for the simultaneous evaluation of these three bioactive compounds in R. rubescens. OBJECTIVE: To develop an optimised ultrasound-assisted extraction (UAE) method and an efficient HPLC method for the simultaneous evaluation of the three bioactive compounds in R. rubescens. METHODOLOGY: Various parameters that can potentially affect the UAE process were investigated and optimised. HPLC operating conditions were also optimised, and the chromatographic separation was performed on a C(18) -column with an acetonitrile-water gradient as the mobile phase. RESULTS: Validation of the HPLC technique developed showed that the method has good linearity, sensitivity, precision and accuracy. The combined UAE-HPLC method was applied to quantitate the amount of oridonin, oleanolic acid and ursolic acid in the plant sample and exhibited good repeatability (RSD (%) < 3.9), reproducibility (RSD (%) < 5.9), stability (RE (%) < 1.4) and recovery (mean (%) > 92.4). Furthermore, the combined UAE-HPLC method was applied successfully to the extraction and determination of oridonin, oleanolic acid and ursolic acid content in samples obtained from different geographical regions, which is the first time that this comparison has been investigated. CONCLUSION: The combined UAE-HPLC process is a fast, convenient and appropriate method for the quantitative analysis of these three compounds in R. rubescens.