Extraction Solvents Ethanol Research Articles

Design of experiment for extraction optimization and antioxidant assay studies of biologically active compounds of propolis

Bees gather propolis from tree sap and combine it with their saliva to seal and sanitize the hive. There are various countries where propolis extracts are used in food and beverage as additives mainly due to the presence of phenolic compounds. Therefore, extraction is the initial and the most vital step in the purification and recovery of phenolic compounds from propolis. The purpose of this study is to enhance propolis extract yield by thoroughly optimizing the extraction process and to evaluate and compare the antioxidant assay of the propolis extracts. Several factors such as the chemical nature of phenolic compounds, composition of solvents, temperature and solvent pH may influence the extraction efficiency and quantity of phenolic compounds. These factors have been extensively studied by univariant optimization; however, the univariant optimization fails to consider the effect of interaction between variables on extraction efficiency. Therefore, the Design of Experiment (DoE) approach is adopted in this study in order to evaluate the correlation between various parameters and their impacts on the extraction of propolis in order to determine TPC, TFC, and DPPH scavenging activity. The DoE was carried out to correctly optimize different extraction parameters including the percentage of ethanol in extraction solvent, temperature and time. The ethanol percentage showed the highest F ratio and minimum p-value for the extraction of 28 total phenolic compounds and flavonoids with a negative correlation showing the presence of more polar compounds in propolis. While in the case of DPPH scavenging assay, the temperature was found to have a significantly positive effect on the extraction efficiency. Optimized extraction parameters obtained by the design were found to be different for TFC, TPC and DPPH, e.g., optimum solvent composition for TFC, TPC and DPPH were 51%, 50% and 90%, respectively. The ultrasonic extracts of propolis showed a higher TPC (574.017 µg GAE/g); in 51% ethanol/water at 25 °C while sonicated for only 2 min. The total flavonoid content (TFC) was found higher (353.16 µg Quercetin/g) in 50% ethanol/water when sonicated for 60 min at 56 °C. DPPH scavenging assay showed higher capacity (1617.27 µg Ascorbic acid/g) in 90% ethanol/water at 62 °C for 60 min sonication. Fourteen propolis samples from Pakistan and Turkey were extracted in order to compare TPC, TFC, and DPPH assay results. In Pakistani propolis, the TPC, TFC, and DPPH scavenging assay varies from 418–1026 µg/g, 264–866 µg/g, and 302–1162 µg/g, respectively. Turkish propolis was demonstrated to have higher TPC, TFC, and DPPH scavenging assay values than Pakistani propolis, with values of 1312–1535 µg/g, 1182–1446 µg/g, and 885–2756 µg/g, respectively. It was found that propolis produced naturally possesses more potential phenolic compounds than propolis purchased from farmer’s markets.

Optimization of extraction, separation and purification of baicalin in Scutellaria baicalensis using response surface methodology

Optimizing extraction, separation, and purification processes of bioactive ingredients improves resource utilization and reduces environmental pollution in pharmaceutical industry. Here, we report findings of a study focusing on enhancing baicalin production from Scutellaria baicalensis, an important medicinal crop with increasing market demand. Using response surface methodology, we optimized ultrasonic-assisted extract method (ethanol concentration, ultrasonic temperature, ultrasonic time) for dried powder and decocting method (liquid-solid ratio, decocting time, decocting times) for fresh slices, respectively. Baicalin was separated, purified, and refined through the processing of macroporous resin, alkali-solution, acid precipitating and methanol reflux, respectively, and was quantified via high performance liquid chromatography. The main findings were as follows: (1) Optimal extraction of dried-powder: particle size 80 mesh, liquid-solid ratio 20:1, extraction solvent ethanol 57%, ultrasonic temperature 68°C, ultrasonic time 66 min. (2) Optimal extraction of fresh slices: adding the fresh slices into boiling water, liquid-solid ratio 43:1, decocting for 93 min. (3) Optimal separation: AB-8 macroporous resin, diameter to height ratio 1:15, loading liquid baicalin concentration 25.6 mg/mL, flow rate 1 BV/h, loading volume 8 BV, and elution volume 11 BV. (4) Optimal purification: adjusting the extraction solution pH to 1.0, static placing at 80°C for 12 h, washing the precipitate twice, dissolving the crude baicalin extract with alkali NaOH, and re-precipitating with stronger acid HCl. (5) Optimal refinement: reflux extracting the above baicalin extraction with 75% methanol for 30 min at liquid-solid ratio 15:1. Collectively, these findings provide valuable references for optimizing processing conditions of important medicinal crops in pharmaceutical industry.

A comparative study on the extraction of betalain rich phenolic compounds in cactus pear

In recent years, consumer demands grow on non-artificial food colorants because of health concerns. Betalain is one of the most popular natural food colorant. The present study concentrated on the extraction of betalain rich bioactive compounds from cactus pear (Opuntia ficus-indica L.). For this purpose, conventional extraction (CE) and ultrasound assisted extraction (UAE) was compared in terms of some extraction parameters including extraction time (x1), extraction temperature (x2), percentage of ethanol in extraction solvent (x3), solid/solvent ratio (x4). Response surface methodology was used in conjunction with Box-Behnken design to optimize the extraction parameters. In CE, the optimum extraction conditions were found as; x1=2.05 h, x2= 50°C, x3=80% and 1/x4= 22.60; optimal conditions were determined as; x1=30 min, x2= 49.99°C, x3=40% and 1/x4= 30 in UAE. Total betalain contents were 418.829 and 471.818 mg betalain/ kg dry matter, respectively in CE and UAE in optimum extraction conditions. It was found that bioactive compounds were extracted effectively in shorter time and lower solvent in UAE. According to results, UAE was superior method for extraction of betalains compared to CE.

Effect of different extraction methods on antioxidant properties and encapsulation efficiency of anthocyanin of pomegranate peel.

This study aimed to measure the efficiency, total anthocyanin content (TAC), and total phenol content (TPC) of pomegranate peel powder (PPP) extract from different extractions. Also, the characteristics of the nanoencapsulated extracts with maltodextrin (MD)/Lepidium perfoliatum (Qodume Shahri) seed gum were investigated. The highest and lowest extraction efficiency was related to solvent ethanol-water extraction (SEWE) (76.35%) and solvent ethanol extraction (SEE) (25.73%), respectively. Extracts obtained from microwave extraction (ME) and ultrasound extraction (UE) methods had the highest and lowest values of TAC (4.00-0.35) (mg C3G/g PPP) and TPC (702.13-232.58) (mg GAE/100 g sample), respectively. Peak 3213 in FT-IR indicates the O-H bond, which showed the highest content of phenolic compounds in the extract obtained from ME compared with SEE, SEWE, and UE. The nanoencapsulated extracts from SEE, SEE, and UE had the lowest particle size of peak 1, particle distribution in peak 1, and average particle size distribution compared with other extractions, respectively. The highest encapsulation efficiency of anthocyanin (EEA) and encapsulation efficiency of phenol (EEP) were related to UE (96.15%) and SEWE (86.57%), respectively. The EEP and EEA of SEE were not significantly different from ME and SEWE, respectively. On the other hand, the type and amount of extractive compounds in the extract have a great impact on the efficiency of nanoencapsulation and the average size distribution of nanoencapsulated particles. As a result, PPP extract is rich in antioxidant compounds, which can be determined by carefully examining the appropriate method of extraction and preservation of the extracted compounds.

In- Vitro Studies on the Antioxidant Activity and Pharmacological Properties of Ciassampelos Pariera L.

The current investigation stated that the antioxidant properties of Cissampelos pareira L. leaf extract of medicinal plants are a rich resource of ingredients that can be used in drug development for human life. The medicinal plant Cissampelos pareira with different solvents such as aqueous, acetone, ethanol, and methanol performed different concentrations like 100, 200, 300, 400, and 500μg/ml were carried out by three methods such as hydrogen peroxide scavenging (H2O2), DPPH and thiobarbituric acid assay. As per the antioxidant properties of C. pareira an aqueous extract of hydrogen peroxide scavenging (H2O2) showed the highest percentage of activity compared to the other methods of antioxidant. The minimum rate of antioxidant properties of DPPH in the C. pareira with 500 μg/ml in high activities than the low concentration. According to the acetone extracts of C. pareira leaf, the maximum percentage of inhibition in hydrogen peroxide scavenging (H2O2) whereas low percentages of inhibition by thiobarbituric acid activity at 500 μg/ml concentration, respectively. Although the extraction solvent ethanol reaction is a concern, the same trend of results of antioxidant properties of hydrogen peroxide scavenging (H2O2) assay showed excellent properties and minimum in thiobarbituric acid was found to be recorded with a maximum concentration of 500 μg/ml respectively. According to the methanolic extract of C. pareira leaf exhibited maximum antioxidant properties at 500μg/ml concentration and minimum percentage of activity at thiobarbituric acid, activity was found to be recorded respectively. Among the three methods, hydrogen peroxide scavenging (H2O2) showed excellent antioxidant activity compared with other methods and ascorbic acid as a standard. Antioxidant activities play a significant role in pharmacological functions because of bioactive ingredients. However, there are some health problems due to the deficiencies of macronutrients that establish an effective manner. This research article describes the current status of the world's herbal medicinal products and properties suggestions for facilitating C. pareira was better tomorrow in humanity.

Chemometric optimisation of pressurised liquid extraction for the determination of alliin and S-allyl-cysteine in giant garlic (Allium ampeloprasum L.) by liquid chromatography tandem mass spectrometry.

Giant garlic is a functional food that contains different kinds of bioactive molecules with beneficial effects on chronic noncommunicable diseases like diabetes and cardiovascular conditions. Considering biosynthesis pathways, abundance, and biological activity, alliin and S-allyl-cysteine were used as chemical markers of organosulphur compounds present in giant garlic. To establish a chemometric optimisation of pressurised liquid extraction for the determination of alliin and S-allyl-cysteine in giant garlic by liquid chromatography tandem mass spectrometry (LC-MS/MS). Samples were blanched (ca. 90°C for 10 min) to inactivate alliinase and γ-glutamyl transpeptidase enzymes and then freeze-dried. Chemometric optimisation was performed via response surface methodology based on central composite design (CCD). Organosulphur compound yields were determined applying a validated LC-MS/MS method in multiple reaction monitoring (MRM) mode using the following transitions: for alliin m/z 178 → 74 and for S-allyl-cysteine m/z 162 → 41. According to CCD results, under constant conditions of pressure (1500 psi) and time (20 min), the optimal conditions for pressurised liquid extraction of alliin and S-allyl-cysteine were 70.75 and 68.97% v/v of ethanol in extraction solvent and 76.45 and 98.88°C as extraction temperature, respectively. Multiple response optimisation for the simultaneous extraction of both organosulphur compounds was established via desirability function. Under these conditions, 2.70 ± 0.27 mg g-1 dry weight (DW) of alliin and 2.79 ± 0.22 mg g-1 DW of S-allyl-cysteine were extracted. These results clearly demonstrated that pressurised liquid extraction is an efficient green technique to extract bioactive organosulphur compounds from giant garlic. Extraction yields were significantly (p < 0.05) higher than those obtained with conventional ultra-turrax extraction.

Astaxanthin Recovery from Shrimp Residue by Solvent Ethanol Extraction Using Choline Chloride:Glycerol Deep Eutectic Solvent as Adjuvant

The present study investigated the use of choline chloride:glycerol deep eutectic solvent as an adjuvant for tuning the ethanol extraction of astaxanthin from Litopenaeus vannamei processing residues. The adjuvant concentration did not show a clear trend on the total carotenoid, however, the addition of adjuvant proved to be advantageous in the ultrasound-assisted experiments. In order to establish a balance between total carotenoid and productivity, an operational condition with ultrasound, 5% (m/v) choline chloride:glycerol and 10 min of incubation was selected, which provided 737.69 μg g-1 total carotenoids and 32.71 μg g-1 astaxanthin. When compared to the astaxanthin standard, the shrimp residue extract obtained greater antioxidant activity in free radical scavenging tests. In addition, the shrimp residue extract dramatically reduced the dosage of norfloxacin (up to 87.5%) and gentamicin (up to 75.0%) antibiotics in tests to inhibit the growth of gram-negative bacteria.

Extraction and Analyses of Flavonoids and Phenolic Acids from Canadian Goldenrod and Giant Goldenrod

Invasive alien plant species Canadian goldenrod (Solidago canadensis L.) and giant goldenrod (Solidago gigantea Aiton) were investigated as a source of phytochemicals and yellow dyes. Flavonoids and phenolic acids were extracted from the inflorescence of Canadian goldenrod with thirteen extraction solvents ethanol, methanol, acetone, water, and mixtures of organic solvents (70%, 80%, and 90%) with water. High performance thin-layer chromatography (HPTLC) coupled to densitometry and high-performance liquid chromatography with photo-diode array detector (HPLC-PDA) were used for analyses of the obtained sample test solutions (STSs), which showed the best and comparable extraction efficiencies for 70% acetone(aq), 70% methanol(aq), and 70% ethanol(aq). HPTLC combined with image analyses in fluorescent mode resulted in different chromatographic fingerprints for Canadian goldenrod and giant goldenrod STSs (70% acetone(aq)) after development, after post-chromatographic derivatization with NP reagent and after use of PEG reagent. The developed HPLC methods enabled analyses of phenolic acids and flavonoids (aglycones and glycosylated) in STSs and hydrolyzed STSs form inflorescence of Canadian and giant goldenrod. Different contents of chlorogenic acid, rutin, hyperoside, isoquercetin, and quercetin were observed in STSs of both goldenrod species. The analyses of hydrolyzed STSs confirmed that glycosylated flavonoids in Canadian and giant goldenrod inflorescence are mainly glycosides of quercetin, kaempferol, and isorhamnetin. Additional analyses using HPTLC and HPLC coupled to tandem mass spectrometry (MS/MS; HPTLC-MS/MS and LC-MS/MS) enabled tentative identification of phenolic acids and flavonoids (10 with HPTLC-MS/MS and 15 with LC-MS/MS), from which several were identified in Canadian (4 with HPTLC-MS/MS and 8 with LC-MS/MS) and in giant (7 with HPTLC-MS/MS and 9 with LC-MS/MS) goldenrod for the first time.

Development of ANN models based on combined UV-vis-NIR spectra for rapid quantification of physical and chemical properties of industrial hemp extracts.

The aim of this study was to develop artificial neural network (ANNs) models for prediction of physical (total dissolved solids, extraction yield) and chemical (total polyphenolic content, antioxidant activity) properties of industrial hemp extracts, prepared by two different extraction methods (solid-liquid extraction and microwave-assisted extraction) based on combined UV-VIS-NIR spectra. Spectral data were gathered for 46 samples per extraction method. The PCA analysis ensured efficient separation of the samples based on the amount of ethanol in extraction solvent using NIR spectra for both conventional and microwave-assisted extraction. Results showed that reliable ANN models (R2 >0.7000) for describing physical, chemical, and simultaneously physical and chemical characteristics can be developed based on combined UV-VIS-NIR spectra of industrial hemp extracts without spectra pre-processing.

Comparative evaluation of the impact of extraction solvent and time on the yield and antioxidant potential of Pinus densiflora needle and bark extracts

This study was aimed to determine and compare the effect of different percentages of extraction solvent ethanol 0% (E0), 20% (E20), and 40% (E40) with water (vol/vol) as well as extraction time 6, 9 and 15 h on the yield and antioxidant potential of Pinus densiflora needle (PDN) and bark (PDB) extracts. Extraction was performed at 60 ± 5 °C. A maximum yield of 10.99 ± 0.96 and 16.86 ± 1.96% from 20 g of PDN and PDB was obtained with E40 at 15 h of extraction, respectively. Extraction of PDN and PDB using E20 or E40 for 6–15 h significantly (P < 0.001) enhanced their free radical scavenging potential compared to the respective E0 extracts. Similarly, the extraction of PDN using E20 or E40 for 6–9 h significantly (P < 0.05 or P < 0.01) improved the oxygen radical absorbance capacity (ORAC) compared to their E0 extracts. However, E20 and E40 did not considerably alter the ORAC of PDB extracts. Additionally, at a minimum concentration of 3.75 μg/ml, all the E20 and E40, but not E0 extracts of PDB showed a significant (P < 0.001) cytoprotective effect against tert-butyl hydroperoxide (TBHP)-induced oxidative stress-mediated cell death. At this minimum concentration, none of the PDN extracts showed such protective activity. Further, it was evident that extraction time significantly altered only PDB-E40 extract yield and antioxidant potential of both PDB and PDN extracts in a solvent-dependent manner. Finally, this study revealed that PDB has a more significant antioxidant potential than PDN.

Chemical components from different parts of Forsythia suspensa vahl with different extraction methods by gas-chromatography-mass spectrometry

With different extraction methods, chemical components from the branch, root and bark of Forsythia suspensa (Thunb.) Vahl were analyzed by gas chromatography-mass spectrometry (GC-MS). The result showed that there were a large number of chemical components in Forsythia suspense, in which there were also different components with different extraction solvent ethanol and benzene/ethanol, respectively. Exacted by ethanol vs. benzene/ethanol, the branch had 79 peaks and 55 chemical components in ethanol, and 31 peaks and 17 components in benzene/ethanol, respectively. The root had 32 peaks and 23 chemical components in ethanol, and 38 peaks and 25 components in benzene/ethanol. As to bark, there were 54 peaks and 39 components with ethanol, 25 peaks and 14 components with ethanol. There were also some common components in different parts with different solvent. It could be found that there were 3 and 7 common extracts in the branch, root and bark Forsythia suspense exacted by ethanol and benzene/ethanol, respectively. It was also found that the same part of Forsythia suspense could have common with different extracted methods. Extracted by ethanol and benzene/ ethanol, the branch, root and bark of Forsythia suspense had 5, 7 and 12 common components, respectively.

Optimization of ultrasonic circulating extraction of samara oil from Acer saccharum using combination of Plackett–Burman design and Box–Behnken design

In this study, ultrasonic circulating extraction (UCE) technique was firstly and successfully applied for extraction of samara oil from Acer saccharum. The extraction kinetics were fitted and described, and the extraction mechanism was discussed. Through comparison, n-hexane was selected as the extraction solvent, the influence of solvent type on the responses was detailedly interpreted based on the influence of their properties on the occurrence and intensity of cavitation. Seven parameters potentially influencing the extraction yield of samara oil and content of nervonic acid, including ultrasound irradiation time, ultrasound irradiation power, ultrasound temperature, liquid-solid ratio, soaking time, particle size and stirring rate, were screened through Plackett–Burman design to determine the significant variables. Then, three parameters performed statistically significant, including liquid-solid ratio, ultrasound irradiation time and ultrasound irradiation power, were further optimized using Box–Behnken design to predict optimum extraction conditions. Satisfactory yield of samara oil (11.72±0.38%) and content of nervonic acid (5.28±0.18%) were achieved using the optimal conditions. 1% proportion of ethanol in extraction solvent, 120°C of drying temperature and 6.4% moisture were selected and applied for effective extraction. There were no distinct differences in the physicochemical properties of samara oil obtained by UCE and Soxhlet extraction, and the samara oil obtained by UCE exhibited better antioxidant activities. Therefore, UCE method has enormous potential for efficient extraction of edible oil with high quality from plant materials.

Experimental and modeling studies on microwave-assisted extraction of mangiferin from Curcuma amada.

Mangiferin, a bioactive compound having potent nutraceutical, strong antioxidant and pharmacological significance has been extracted using microwave-assisted extraction (MAE) technique from Curcuma amada, commonly known as mango ginger. The extraction solvent ethanol is eco-friendly, nontoxic and reduces the risk of environmental hazards. The influence of several independent variables such as microwave power, ethanol concentration, extraction (irradiation) time and pre-leaching time has been studied under varying conditions using one-factor-at-a-time analysis to obtain an optimal extraction ratio. The maximum mangiferin content of 1.1156 mg/g is obtained at microwave power of 550 W and extraction time of 50 s with 80 % ethanol as a solvent and pre-leaching time of 20 min. The results indicate that microwave power and ethanol concentration have the most significant effect on the yield of mangiferin content. The presence of mangiferin in final Curcuma amada extract is confirmed through high-performance liquid chromatography and the functional groups are identified through Fourier transform infrared spectroscopy analyses using standard mangiferin. The experimental profiles are fitted into a two-parameter modified first-order kinetic model and a three-parameter modified logistic model and checked using the goodness-of-fit criterion. The Curcuma amada retained its antioxidant activity after MAE treatment and the antioxidant activity of mangiferin obtained after extraction using DPPH free radical scavenging assay is studied.

Optimization of the Ocimum basilicum L. extraction process regarding the antioxidant activity

The levels of input variables (temperature and extraction solvent) that optimize a particular response (total phenols content, total flavonoids content and antioxidant activity) of the Ocimum basilicum L. extraction process were determined by the response surface methodology (RSM). The influence of theextraction temperature on extraction process was investigated in the range from 33.8?C to 76.2?C, as well as of extraction solvent ethanol, in the range of concentrations from 21.7% to 78.3%. For the preparation of basil dry extract, characterized with minimal IC50 value, the calculated optimal values of temperature and ethanol concentration were: 75.33?C and 73.66% (w/w).

Iberis amara L. constituents reduce excitatory cholinergic and inhibitory purinergic neurotransmission

An extract of Iberis amara L. (extraction solvent ethanol 50%, 1:1.5–2.5) is, together with eight other herbal extracts, component of STW 5 (Iberogast®), a phytomedicine with clinically proven efficacy and safety in functional gastrointestinal diseases (1, 2). The objective of this study was to evaluate the pharmacological effects of Iberis amara and some if its characteristic constituents (glucoiberin, kaempferol, cucurbitacin E and I, and kaempferol-3,4′-diglucopyranoside-7-O-rhamnopyranoside) on cholinergic neurotransmission in the enteric nervous system of rat and mouse. Cholinergic contractions of isolated muscle strips of ileum were studied in an organ bath. Myenteric reflex responses elicited by EFS were studied in a perfused organ bath using a four inch long ileum segment. Intracellular recordings of EJP and IJP in smooth muscle cells of the circular muscle layer of the proximal colon were performed. Iberis amara and its components kaempferol and cucurbitacin E reduced cholinergic contractions in organ bath studies in a concentration dependent manner. Kaempferol and the cucurbitacins E and I reduced the amplitude and latency of ascending and descending myenteric reflex contractions in a concentration dependent manner. Kaempferol reduced cholinergic EJP and increased the purinergic fast part of the IJP. Thus, Iberis amara and its characteristic components interact with mechanisms of excitatory cholinergic and inhibitory purinergic neurotransmission in the enteric nervous system. Characterization of these distinct effects on gastrointestinal neurotransmission helps to understand how the combination phytomedicine STW 5 alleviates patient symptoms.

Insect Antifeedant Sesquiterpene Acetals from the Liverwort Lepidolaena clavigera. 2. Structures, Artifacts, and Activity

Clavigerin A ( 1) was isolated from the New Zealand liverwort Lepidolaena clavigera and shown to be a polyoxygenated bergamotane sesquiterpene with an unusual ring system. L. clavigera shows infraspecific variation, since 1 was the only clavigerin detected in a North Island collection, whereas the previously reported clavigerins B ( 2) and C ( 3) were found in South Island collections with no sign of 1. Three new clavigerins, 4- 6, were identified, but these are artifacts formed by alcoholysis of the acetoxy acetal group of the clavigerins 2 and 3, with either the extraction solvent ethanol or the RP column eluent methanol. The insect antifeedant and cytotoxic activities of these compounds are reported, and it is proposed that they act as hidden 1,4-dicarbonyl compounds.