Radical-scavenging activity of extracts from Cordia verbenacea DC obtained by different methods

The possibility of using the leaves of Cordia verbenacea as a new source of natural antioxidant compounds was investigated. In the present work, extracts from C. verbenacea were obtained using different extraction methods: supercritical fluid extraction (SFE), Soxhlet (SE), hydrodistillation and maceration, with the objective to evaluate the methods in terms of yield and antioxidant potential. The high-pressure technique was applied using pure CO₂ and CO₂ with co-solvent at different temperatures and pressures (30, 40 and 50 °C and 100, 200, and 300 bar). Organic solvents with different polarities were used to obtain extracts by low-pressure extraction processes. The extracts were evaluated according to their antioxidant activity using total phenolic content, scavenging abilities on DPPH radical, total antioxidant activities (ABTS^•+), superoxide anion radical-scavenging (O₂^•) and protection against lipid peroxidation in vitro (LPO). Ethyl acetate fraction obtained by maceration and extract isolated by SE using 25% aqueous mixture of ethanol possessed the highest scavenger activity against DPPH radical (IC₅₀ = 9.2 ± 0.4 μg/ml, IC₅₀ = 27.4 ± 0.1 μg/ml, respectively). The SFE with 8% ethanol as a co-solvent produced extracts with distinguished increase in the antioxidant activity. The Soxhlet extract with ethyl acetate exhibited a strong reduction of lipid peroxidation (IC₅₀ = 209 ± 3 μg/ml) value comparable to the standard rutin (IC₅₀ = 203 ± 2 μg/ml). The results indicate that extracts of C. verbenacea have important potential as a source of bioactive compounds with antioxidant activity.     

SFE from Bidens pilosa Linné to obtain extracts rich in cytotoxic polyacetylenes with antitumor activity

Bidens pilosa L. is a plant considered medicinal by some South American cultures. It contains polyacetylenes which may be the constituents responsible for its antitumor activity. Extracts obtained by hydroethanol maceration (HCE) and supercritical fluid extraction (SFE) were monitored for antitumor activity and the presence of polyacetylenes in the constitution. Both extracts killed concentration-dependently the MCF-7 cells in culture, although the SFE extract presented superior cytotoxic activity. The SFE presented IC₅₀ = 437 (428-446) μg/mL in 24 h of incubation, decreasing to IC₅₀ = 291 (282-299) μg/mL at 48 h. The HCE started causing DNA cleavage at 160 μg/mL while the SFE extract started at 40 μg/mL, a concentration enough to initiate the in vitro cleavage. The presence of polyacetylenes as the major compounds in SFE was confirmed by TL chromatography combined with UV-vis analyses. Ehrlich ascites carcinoma-bearing mice were used for the antitumor study. Animals were divided in five groups: normal, negative control, positive control (Doxorubicin 0.06 mg/kg), test group HCE and test group SFE (100 mg/kg b.w. per day). After 9 days of treatment, 50% of randomly chosen animals from each group were sacrificed for the study. The parameters evaluated were: body weight, abdominal circumference, volume of ascitic fluid and tumor cells, viable and nonviable tumor cell count, determination of mean survival time and increased life-span. Both extracts presented antitumor activity, but SFE reduced more the volumes of ascites fluid and the tumor cells (4 ± 1 and 1 ± 0.4 mL, respectively), while caused higher mean survival time (17 days) and increased life span (∼31%). The results suggest the importance of the polyacetylenes from B. pilosa as leader molecules to contribute to a new anticancer drug by using the supercritical technology.     

Processing cherries (Prunus avium) using supercritical fluid technology. Part 1: Recovery of extract fractions rich in bioactive compounds

In the recent years many studies on cherries revealed that they are rich sources of bioactive compounds with beneficial biological activity.In this work, fractioned high pressure extractions were performed on a traditional sweet cherry variety from Portugal (“Saco”) in order to recover natural ingredients with bioactivity. The methodology employed comprised a first step with supercritical CO₂ followed by a second step where different mixtures of CO₂ and ethanol (10-100%, v/v) were tested. All extractions were performed at 50 °C and 25 MPa during 1 plus 1.5 h and the resulting extracts were then characterized in terms of global yield, phenolic content, antioxidant activity and screened for the presence of perillyl alcohol, a powerful anticancer compound. Moreover, cell-based assays were also performed as preliminary evaluation of potential antiproliferative activity of the cherry extracts.In the first step, lower yields were obtained and the extracts presented lower phenolic and antioxidant activity than those recovered in the second step. The extract obtained with CO₂:EtOH (90:10, v/v) exhibited the highest antioxidant activity (181.4 ± 23.7 μmol TEAC/g) and was the most effective in inhibiting the growth of human colon cancer cells (ED50_96h = 0.20 ± 0.02 mg/mL). Perillyl alcohol was pointed to be one of the major responsible for antiproliferative properties of cherry extracts as it was detected in the most promising products, and polyphenols, in particular sakuranetin and sakuranin, seemed to be the major contributors of the antioxidant capacity.     

Effect of solvent (CO2/ethanol/H2O) on the fractionated enhanced solvent extraction of anthocyanins from elderberry pomace

The management of agro-industrial residues is an important issue for environmental reasons and the reuse of byproducts represents a good alternative, especially if it is conjugated with green technologies and the production of valuable products. Portuguese elderberry pomace is rich in anthocyanins with therapeutic properties that confer to this byproduct potential to be applied in the food and pharmaceutical industries. Fractionated high pressure extractions from elderberry pomace were performed using supercritical CO₂ extraction, followed by enhanced solvent extraction (ESE) with diverse CO₂/ethanol/H₂O solvent mixtures (0-90%, 0.5-100%, 0-95%, v/v/v), at 313 K and 21 MPa, in order to obtain anthocyanin-rich fractions. The ESE solvent mixtures had a substantial effect on extracts yield and composition. The maximum extraction yield (24.2%), total phenolic compounds (15.8% gallic acid equivalents), total flavonoids (8.9% epicatechin equivalents), total anthocyanins (15.0% cyanidin-3-glucoside equivalents) and antioxidant activity (IC₅₀ of 21 μg) achieved highlight the great potential of elderberry pomace for valuable applications.     

Supercritical fluid extraction of flavonoids from Maydis stigma and its nitrite-scavenging ability

Supercritical fluid carbon dioxide (SF-CO₂) extraction (SFE) of flavonoids from Maydis stigma and its nitrite-scavenging ability were investigated. The effects of extraction time, particle size and co-solvent composition in terms of water content in ethanol were first optimized. Then, a Box-Behnken design combined with response surface methodology (RSM) was employed to study the effects of three independent variables (temperature, pressure and co-solvent amount) on the extraction yield of flavonoids. A maximal extraction yield of flavonoids of approximately 4.24 mg/g of M. stigma by SFE was obtained under optimal conditions (a temperature of 50.88 °C, a pressure of 41.80 MPa, a co-solvent amount of 2.488 mL/g and an extraction time of 120 min with 0.4-mm particle sizes and 20% aqueous ethanol as the co-solvent). Furthermore, the nitrite-scavenging ability of the flavonoid-enriched SFE extracts was assessed using the Griess reagent. The flavonoid-enriched SFE extracts exhibited the highest scavenging ability on nitrite (88.1 ± 3.04%) at the concentration of 500 μg/mL and at pH 3.0. The nitrite-scavenging ability of the extracts appeared to be concentration dependent but negatively correlated with the pH.     

Chemical composition, angiotensin I-converting enzyme inhibitory, antioxidant and antimicrobial activities of essential oil of Tunisian Thymus algeriensis Boiss. et Reut. (Lamiaceae)

The present study describes chemical composition, angiotensin I-converting enzyme (ACE) inhibitory, antioxidant and antimicrobial activities of the essential oil of wild growing Thymus algeriensis Boiss. et Reut. (Lamiaceae), a traditional medicinal plant which is mainly endemic in Tunisia and Algeria. The essential oil from the fresh leaves and flowers of T. algeriensis were extracted by hydrodistillation and analysed by GC and GC/MS. Fifty-seven compounds were identified accounting for 97.71% of the total oil, where oxygenated monoterpenes constituted the main chemical class (44.85%). The oil was dominated by camphor (7.82%), 4-terpineol (7.36%), α-pinene (6.75%), 1,8-cineole (5.54%) and cis-sabinene hydrate (5.29%). The T. algeriensis essential oil was found to possess an interesting inhibitory activity towards ACE with an IC50 value of 150 μg/ml. The obtained results also showed that this oil can act as radical scavengers (IC₅₀ = 0.8 mg/ml) and displayed a lipid peroxidation inhibitory activity (IC₅₀ = 0.5 mg/ml) as evaluated by 2,2-diphenyl-1-picrylhydrazyl and β-carotene bleaching methods, respectively. Furthermore, the oil was tested for antimicrobial activity against six bacterial strains and two fungal strains. The inhibition zones and minimal inhibitory concentration values of microbial strains were in the range of 13.5-64 mm and 1-6 μl/ml, respectively. The oil exhibited remarkable inhibitory activity against fungal and Gram-positive bacteria strains.     

Brazilian Ginseng extraction via LPSE and SFE: Global yields, extraction kinetics, chemical composition and antioxidant activity

Brazilian Ginseng extracts of two species, Pfaffia paniculata and Pfaffia glomerata, were obtained by supercritical fluid extraction (SFE) with CO₂ and by low-pressure solvent extraction (LPSE) with methanol, hexane and ethanol. The SFE assays were conducted at pressures of 100, 200 and 300 bar, and temperatures of 30 and 50 °C. The qualitative chemical compositions of the extracts were determined by thin layer chromatography (TLC). One of the active principles of interest from P. glomerata extract, β-ecdysone, was identified and quantified by HPLC. The antioxidant activities of Brazilian Ginseng extracts were determined by the coupled reaction of linolenic acid and β-carotene. For P. paniculata, the highest SFE yield was obtained at 200 bar/50 °C (0.22%, dry basis—d.b.), while the best extraction condition for P. glomerata was obtained at 200 bar/30 °C (0.18%, d.b.). The higher extract yields obtained by LPSE were 2.0% and 5.8% (w/w, d.b.) for P. paniculata and P. glomerata, respectively, both obtained with methanol as extraction solvent. From the overall extraction curve of P. glomerata, it was possible to obtain the kinetic parameters of extraction; the duration of the CER (constant extraction rate) period was determined as 134 min. The TLC plates showed the possible presence of flavonoids in the ethanolic extract for both Pfaffia species. The antioxidant activity analysis detected that LPSE extracts had higher activity than SFE extracts.     

Supercritical fluid extraction of lycopene from tomato juice and characterization of its antioxidation activity

A new method to extract lycopene from tomato juice using supercritical CO₂ as solvent and without the need to dry the raw material is presented. To conduct the extraction, the tomato juice was subjected to cycles of centrifugation followed by rinsing with absolute ethanol to partially remove the water present in the solid part of the juice. The influence of the temperature and pressure on the extraction efficiency and on the extract antioxidant activity was studied using a factorial experimental design. The extraction efficiency varied from 7.7% to 76.7% and only extraction temperature had a statically significant effect on the process. The reversed phase HPLC analysis showed that lycopene is the major compound of the extract. The extract that presented higher antioxidant activity was obtained at 40 °C and 350 bar with 12.7 mmol L⁻¹ Trolox/g of extract using the DPPH radical scavenging method and 61.3 mmol L⁻¹ Trolox/g of extract using the rubrene singlet oxygen quenching method.     

Extraction of phenolic compounds from pitanga (Eugenia uniflora L.) leaves by sequential extraction in fixed bed extractor using supercritical CO2, ethanol and water as solvents

With the goal of maximizing the extraction yield of phenolic compounds from pitanga leaves (Eugenia uniflora L.), a sequential extraction in fixed bed was carried out in three steps at 60 °C and 400 bar, using supercritical CO₂ (non-polar) as solvent in a first step, followed by ethanol (polarity: 5.2) and water (polarity: 9.0) in a second and third steps, respectively. All extracts were evaluated for global extraction yield, concentration and yield of both polyphenols and total flavonoids and antioxidant activity by DPPH method (in terms of EC₅₀). The nature of the solvent significantly influenced the process, since the extraction yield increased with solvent polarity. The aqueous extracts presented higher global extraction yield (22%), followed by ethanolic (16%) and supercritical extracts (5%). The study pointed out that the sequential extraction process is the most effective in terms of global extraction yield and yield of polyphenols and total flavonoids, because it produced the more concentrated extracts on phenolic compounds, since the supercritical ethanolic extract presented the highest phenolics content (240.5 mg GAE/g extract) and antioxidant capacity (EC₅₀ = 9.15 μg/mL). The most volatile fraction from the supercritical extract, which is similar to the essential oils obtained by steam distillation or hydrodistillation, presented as major compounds the germacrenos D and B + bicyclogermacrene (40.75%), selina-1,3,7(11)-trien-8-one + selina-1,3,7(11)-trien-8-one epoxide (27.7%) and trans-caryophyllene (14.18%).     

Antioxidant Activity of Alhagi camelorum Phenolics Extracted by Automated and Standard Extraction Techniques

Two extraction techniques, automatic soxtherm apparatus and conventional ultrasonic extraction, were compared in terms of yield, composition, and with special focus on antioxidant activity. In the DPPH test automated extraction showed higher antioxidant potential with IC₅₀ value of 7.5 mg/mL as compared to that of ultrasonic extraction which is 12.5 mg/L, whereas combining both was found to be relatively better. A similar trend in total phenolic and total flavonoid contents was observed. HPLC analysis of both extracts revealed the presence of gallic acid, p-hydroxybenzoic acid, caffeic acid, syringic acid rutin, and ferrulic acid with significant difference in their profile.     

Efficient and selective extraction of magnolol from Magnolia officinalis by mechanochemical extraction technique

Mechanochemical extraction technique was explored for efficient and selective extraction of magnolol from Magnolia officinalis. This process was carried out using a high intensive activator, AGO-2. The yield of magnolol via mechanochemical method was maximized under the following conditions: milling with Na₂CO₃ (2.0 wt%) for 7 min; extraction by water at 40 °C and liquid/solid ratio of 25:1 mL/g for 20 min; precipitation at pH 3.5. The results from scanning electron micrographs, infra-red spectrum and nuclear magnetic resonance indicate that mechanochemical action could result in the disruption of cell walls and the transformation of magnolol into a water-soluble salt form with the addition of sodium carbonate. Compared with superfine grinding extraction and heat-reflux extraction, mechanochemical extraction method reduced both extraction time and temperature, and also achieved a higher magnolol yield and content in the crude extracts.     

Selective Extraction of Piceatannol from Passiflora edulis by-Products: Application of HSPs Strategy and Inhibition of Neurodegenerative Enzymes

Passiflora edulis by-products (PFBP) are a rich source of polyphenols, of which piceatannol has gained special attention recently. However, there are few studies involving environmentally safe methods for obtaining extracts rich in piceatannol. This work aimed to concentrate piceatannol from defatted PFBP (d-PFBP) by means of pressurized liquid extraction (PLE) and conventional extraction, using the bio-based solvents selected with the Hansen solubility parameters approach. The relative energy distance (Ra) between solvent and solute was: Benzyl Alcohol (BnOH) < Ethyl Acetate (EtOAc) < Ethanol (EtOH) < EtOH:H₂O. Nonetheless, EtOH presented the best selectivity for piceatannol. Multi-cycle PLE at 110 °C was able to concentrate piceatannol 2.4 times more than conventional extraction. PLE exhibited a dependence on kinetic parameters and temperature, which could be associated with hydrogen bonding forces and the dielectric constant of the solvents. The acetylcholinesterase (AChE) and lipoxygenase (LOX) IC₅₀ were 29.420 μg/mL and 27.682 μg/mL, respectively. The results reinforce the demand for processes to concentrate natural extracts from food by-products.     

Extraction of flavonoids and carotenoids from Thai silk waste and antioxidant activity of extracts

Extraction of carotenoids and flavonoids from yellow Thai silk waste was investigated. The total recovery of 0.7 mg carotenoids and 5.1 mg flavonoids/g dry weight was obtained by ethanol extraction. Different methods for extractions of these pigments were carried out using two benign solvents: ethanol and subcritical water (SW) to determine the extraction efficiency of the solvents in various extraction conditions. For extraction of carotenoids, ethanol was suitable as extraction solvent and the amount of carotenoids increased with increasing temperature and extraction time. For flavonoids, SW extraction was suitable but the amount of flavonoids decreased with increasing SW temperature and extraction time due to decomposition at such conditions. In addition, the silk extracts were found to have low IC₅₀ values (15.6–23.3 μg/ml), the concentrations of the silk extracts that exhibit 50% reduction in 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), (ABTS) free radicals, thus indicating high antioxidant activities.     

Combination of supercritical fluid extraction with dispersive liquid-liquid microextraction for extraction of organophosphorus pesticides from soil and marine sediment samples

Supercritical fluid extraction (SFE) coupled with dispersive liquid-liquid microextraction (DLLME) and followed by gas chromatography-flame ionization detection (GC-FID) was applied for extraction and determination of ultra-trace amounts of seven organophosphorus pesticides (OPPs) (o,o,o-triethyl phosphorothioate, thionazin, sulfotepp, disulfoton, methyl parathion, parathion, and famphur) in soil and marine sediment samples. Supercritical CO₂ at 150 bar, 60 °C, 10 min static and 30 min dynamic extraction times was used to extract the pesticides. The extracts were collected in 1.0 mL of acetonitrile. Seventeen μL of carbon tetrachloride was dissolved in the collecting solvent and the mixture was then injected rapidly into 5.0 mL of aqueous solution. About (5.0 ± 0.2 μL) of sediment phase was collected after centrifuging and finally 2.0 μL of it was injected into gas chromatography (GC) injection port for analyses. The extraction recoveries for the target analytes were in the range of 44.4% and 95.4% and relative standard deviation (RSD%) for four-replicate measurements was below 7.5%. The limit detections of the method for determining the pesticides were in the range of 0.001-0.009 mg kg⁻¹. The method was successfully applied for analysis of OPPs in real soil and marine sediment samples and satisfactory results were obtained.     

Inhibition of 12- and 15-lipoxygenase activities and protection of human and tilapia low density lipoprotein oxidation by I-Tiao-Gung (<Emphasis Type="Italic">Glycine tomentella</Emphasis>)

I-Tiao-Gung, Glycine tomentella, has been used extensively as a traditional herbal medicine to relieve physical pain, but its bioactivity has not been studied systematically. Ninety-five percent ethanol extracts of G. tomentella (GT-E) showed antioxidant activity in human plasma by prolonging the lag phase (+T_lag) of Cu²⁺-induced, LDL oxidation and were dose dependent. The+T_lag of LDL combined with 3.2 μg/mL GT-E was similar to that with 2.0 μM (ca. 0.5 μg/mL) Trolox. A similar inhibitory effect was found toward tilapia plasma LDL. In addition, GT-E inhibited tilapia thrombocyte (nucleated platelet), 5-, 12-, and 15-lipoxygenase (LOX). The IC₅₀ values were 0.43, 0.72, and 0.42 μg/mL, respectively, whereas the IC₅₀ values for nordihydroguaiaretic acid (NDGA) on 5-, 12-, and 15-LOX were 2.3, 1.6, and 1.7 μg/mL, respectively. The IC₅₀ value for cyclooxygenase-2 (COX-2) inhibition by GT-E was 42.0 μg/mL, whereas the IC₅₀ value by indomethacin as a positive control was 0.61 μg/mL. The prevention of LDL oxidation and the dual inhibition of LOX and COX-2 are indicative of the possible roles of I-Tiao-Gung in antiatherosclerosis and anti-inflammation.     

Comparison of different extraction methods for the extraction of major bioactive flavonoid compounds from spearmint (Mentha spicata L.) leaves

Different bioactive flavonoid compounds including catechin, epicatechin, rutin, myricetin, luteolin, apigenin and naringenin were obtained from spearmint (Mentha spicata L.) leaves by using conventional soxhlet extraction (CSE) and supercritical carbon dioxide (SC-CO₂) extraction at different extraction schemes and parameters. The effect of different parameters such as temperature (40, 50 and 60 °C), pressure (100, 200 and 300 bar) and dynamic extraction time (30, 60 and 90 min) on the supercritical carbon dioxide (SC-CO₂) extraction of spearmint flavonoids was investigated using full factorial arrangement in a completely randomized design (CRD). The extracts of spearmint leaves obtained by CSE and optimal SC-CO₂ extraction conditions were further analyzed by high performance liquid chromatography (HPLC) to identify and quantify major bioactive flavonoid compounds profile. Comparable results were obtained by optimum SC-CO₂ extraction condition (60 °C, 200 bar, 60 min) and 70% ethanol soxhlet extraction. As revealed by the results, soxhlet extraction had a higher crude extract yield (257.67 mg/g) comparing to the SC-CO₂ extraction (60.57 mg/g). Supercritical carbon dioxide extract (optimum condition) was found to have more main flavonoid compounds (seven bioactive flavonoids) with high concentration comparing to the 70% ethanol soxhlet extraction (five bioactive flavonoids). Therefore, SC-CO₂ extraction is considered as an alternative process compared to the CSE for obtaining the bioactive flavonoid compounds with high concentration from spearmint leaves.     

A green separation process for recovery of healthy oil from pumpkin seed

In this study, pumpkin seeds, called as “Ürgüp Sivrisi” and grown in Cappadocia region, were used as plant materials because of high aroma contents. In the supercritical fluid extraction of pumpkin seed oil, the effect of main process parameters as the particle size (250-2360 μm), the volumetric flow rate of supercritical solvent (0.06-0.30 L/h), the operating pressure (20-50 MPa), the operating temperature (40-70 °C), the type of entrainer (ethanol and n-hexane) and those concentrations (0-10 vol.%) on the extraction yield, the oil solubility and the initial extraction rate were investigated. A cross-over effect for the extraction of pumpkin seed oil using supercritical CO₂ was determined at the operating pressure of 20-30 MPa. The maximum extraction yield obtained with entrainer free was reached 0.50 g oil/g dry seed at 600-1180 μm, 0.12 L/h, 50 MPa and 70 °C for the operation time of 5 h. The maximum extraction yield obtained with ethanol as an entrainer in the experiments was reached 0.54 g oil/g dry seed at the conditions of 600-1180 μm, 0.12 L/h, 30 MPa, 40 °C and 8 vol.% for the operating time of 2 h. The oil compositions were determined by gas chromatography analysis and the results showed that the compositions of pumpkin seed oil which were obtained by means of organic solvent extraction and supercritical fluid extraction were similar. The average oil compositions determined as 9.3 (±0.43)% palmitic acid, 7.5 (±0.6)% stearic acid, 32.3 (±0.6)% oleic acid, 48.1 (±0.6)% linoleic acid and 0.7 (±0.3)% linolenic acid. The morphological changes in the seeds were determined by the scanning electron microscopy analysis.     

Supercritical fluid extraction of piceid, resveratrol and emodin from Japanese knotweed

In the present study, the use of supercritical fluid extraction was investigated for selected compounds from the plant Japanese knotweed (Polygonum cuspidatum Siebold & Zucc.). The effects of parameters such as type of modifier, pressure, temperature and time on the extraction efficiency of piceid, resveratrol and emodin were studied. The optimal conditions were found as follows: modifier acetonitrile, 40 MPa, 100 °C and 45 min. SFE results were compared with those obtained by conventional Soxhlet extraction carried out for 4 h. The extracts obtained using these two techniques were analysed by liquid chromatography coupled with UV detection. LiChrospher^® 100, RP-18 column (125 mm × 4 mm, 5 μm) coupled with gradient elution acetonitrile in acidified water was used for the separation of compounds at flow rate 0.5 mL min⁻¹. Detection was carried out at 306 nm. Limits of detection were 21, 8 and 52 μg L⁻¹ for piceid, resveratrol and emodin, respectively. The linear range was 0.5-10 mg L⁻¹ for piceid and resveratrol, and 1-50 mg L⁻¹ for emodin with correlation coefficients above 0.9981. Based on the comparison of both methods extracted amount of piceid by Soxhlet extraction is approximately 10 times higher than by SFE method, while the extraction yield of emodin by Soxhlet extraction in approx. 2.5 times lower than by SFE. The advantage of SFE over Soxhlet extraction method is more than 5 times shorter extraction time period.     

Optimization of subcritical water extraction of antioxidants from Coriandrum sativum seeds by response surface methodology

Subcritical water extraction (SWE) of antioxidants from Coriandrum sativum seeds (CSS) was optimized by simultaneous maximization of the total phenolics (TP) and total flavonoids (TF) yield and antioxidant activity, using IC₅₀ value. Box–Behnken experimental design (BBD) on three levels and three variables was used for optimization together with response surface methodology (RSM). Influence of temperature (100–200 °C), pressure (30–90 bar) and extraction time (10–30 min) on each response was investigated. Experimentally obtained values were fitted to a second-order polynomial model and multiple regression. Analysis of variance (ANOVA) was used to evaluate model fitness and determine optimal conditions. Moreover, three-dimensional surface plots were generated from employed mathematical model. The optimal SWE conditions obtained in simultaneous optimization were temperature of 200 °C, pressure of 30 bar and extraction time of 28.3 min, while obtained values of TP and TF yields and IC₅₀ value at this experimental point would be 2.5452 g GAE/100 g CSS, 0.6311 g CE/100 g CSS and 0.01372 mg/ml, respectively. Moreover, good and moderate linear correlation was observed between antioxidant activity (IC₅₀ value) and total phenolics content (R² = 0.965), and total flavonoids content (R² = 0.709) which indicated that these groups of compounds are responsible for antioxidant activity of C. sativum extracts.     

Design,Synthesis, Biological Evaluation, 2D-QSAR Modeling,and Molecular Docking Studies of Novel 1H-3-Indolyl Derivatives as Significant Antioxidants

Novel candidates of 3-(4-(thiophen-2-yl)-pyridin/pyran/pyrimidin/pyrazol-2-yl)-1H-indole derivatives (2–12) were designed by pairing the pyridine/pyrane/pyrimidine/pyrazole heterocycles with indole and thiophene to investigate their potential activities as (2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) inhibitors. The purpose of these derivatives’ modification is to create high-efficiency antioxidants, especially against ABTS, as a result of the efficiency of this set of key heterocycles in the inhibition of ROS. Herein, 2D QSAR modeling was performed to recommend the most promising members for further in vitro investigations. Furthermore, the pharmacological assay for antioxidant activity evaluation of the yielded indole-based heterocycles was tested against ABTS (2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid); by utilizing ascorbic acid as the standard. Candidate 10 showed higher antioxidant activity (IC₅₀ = 28.23 μg/mL) than ascorbic acid itself which achieved (IC₅₀ = 30.03 μg/mL). Moreover, molecular docking studies were performed for the newly designed and synthesized drug candidates to propose their mechanism of action as promising cytochrome c peroxidase inhibitors compared to ascorbic acid as a reference standard. Our findings could be promising in the medicinal chemistry scope for further optimization of the newly designed and synthesized compounds regarding the introduced structure-activity relationship study (SAR) in order to get a superior antioxidant lead compound in the near future.