Response surface methodology for protein extraction optimization of red pepper seed (Capsicum frutescens)

Response surface methodology was used to determine optimum conditions for extraction of protein from red pepper seed meal. A central composite design including independent variables such as temperature (30, 35, 40, 45 and 50 °C), pH (7.0, 7.5, 8.0, 8.5 and 9.0), extraction time (20, 30, 40, 50 and 60 min) and solvent/meal ratio (10:1, 15:1, 20:1, 25:1 and 30:1 v/w) was used. Selected response (dependent variable) which evaluates the extraction process was protein yield and the second-order model obtained for protein yield revealed coefficient of determination of 96.7%. Protein yield was primarily affected by pH and solvent/meal ratio. Maximum yield was obtained when temperature, pH, mixing time and solvent/meal ratio were 31 °C, 8.8, 20 min, 21:1 (v/w), respectively. These conditions resulted in protein yield of 12.24 g of soluble protein from extract/100 g defatted red pepper seed flour. The adequacy of the model was confirmed by extracting the protein under optimum values given by the model. These results help in designing the process of optimal protein extraction from red pepper seeds.     

Production of soy protein isolates with low phytic acid content by membrane technologies: Impact of the extraction and ultrafiltration/diafiltration conditions

The content of the antinutrient, phytic acid, of soy protein was analyzed during their extraction and purification by a series of ultrafiltration and diafiltration steps. The phosphorus content of the extracts was used as an indication of their phytic acid content and their ash content as an indication of their mineral content. The extraction of soy proteins was conducted by using a 2³ factorial experimental design, pH (7.5 or 9), solvent (0.06 M KCl or water), and temperature (25 °C or 50 °C). The most promising extraction conditions were 0.06 M KCl/pH 9.0/25 °C for the lowest phosphorus to protein ratio (12.2 ± 0.1 mg P/g protein) and H₂O/pH 9.0/50 °C for the combination of low phosphorus to protein ratio and the lowest ash content (13.9 ± 1.2 mg P/g protein, 9.6 ± 0.8% w/w ash content). After extraction, soy proteins were purified by sequential ultrafiltration (UF) with a volume concentration ratio (VCR) of 5 and diafiltration (DF) with volume diafiltration ratio (VD) of 4. Extracts were purified with no pH adjustment or with pH adjustment to 6.5 between the UF and the DF steps. The extraction conditions 0.06 M KCl/pH 9.0/25 °C and the purification conditions UF pH 9.0/DF pH 6.5 showed the lowest phosphorus to protein ratio (4.4 ± 0.3 mg P/g protein) and reduced membrane fouling when compared to extraction conditions with water.     

Extraction optimization of watermelon seed protein using response surface methodology

Extraction conditions for maximum protein recovery from watermelon (Citrullus vulgaris Cv Sugar baby) seed meal were investigated using response surface methodology. A central composite design with four independent variables: temperature (40, 45, 50, 55 and 60 °C); NaOH concentration (0.03, 0.06, 0.09, 0.12 and 0.15 g/L); extraction time (5, 10, 15, 20 and 25 min) and solvent/meal ratio (30:1, 40:1, 50:1, 60:1 and 70:1, v/w) was used to study the response variable (protein yield). The experimental values of protein yield ranged between 72.03 and 81.52 g/100 g seed meal. A second-degree equation for independent and response variables was computed and used to create the contour plots graphs. The predicted values obtained for protein yield revealed that coefficient of determination and standard error was 0.80 and 0.906, respectively. Optimum protein extraction was obtained with 0.12 g/L alkali concentration, 15 min extraction time and 70:1 (v/w) solvent/meal ratio at 50 °C. Confirmatory studies revealed that the protein yield under optimum conditions was 80.71 g/100 g seed meal.     

Effect of extraction conditions on lycopene extractions from tomato processing waste skin using response surface methodology

Skin, rich in lycopene, is an important component of waste originating from tomato paste manufacturing plants. A central composite design with five independent variables, namely solvent/meal ratio (20:1, 30:1, 40:1, 50:1, and 60:1 v/w); number of extractions (1, 2, 3, 4 and 5); temperature (20, 30, 40, 50 and 60 °C); particle size (0.05, 0.15, 0.25, 0.35 and 0.43 mm); extraction time (4, 8, 12, 16 and 20 min) was used to study their effects on lycopene extraction. The experimental values of lycopene ranged between 0.639 and 1.98 mg/100 g. The second order model obtained for extracted lycopene revealed a coefficient of determination (R²) of 0.99 and a standard error of 0.03. Maximum lycopene (1.98 mg/100 g) was extracted when the solvent/meal ratio, number of extractions, temperature, particle size and extraction time were 30:1 v/w, 4, 50 °C, 0.15 mm and 8 min, respectively.     

Optimization of lentil protein extraction and the influence of process pH on protein structure and functionality

Response surface methodology was used to optimize alkaline extraction of protein from lentil flour to maximize both protein content and yield. Solid/solvent ratio and pH were the significant factors that determined protein extraction efficiency. At the optimized condition of pH 9.0 and solid/solvent ratio of 1:10 (g:mL), a yield of 14.5 g of protein extract/100 g of flour was obtained with a protein content of 82 g/100 g at (22 °C) after 1 h of extraction. The impact of extraction pH on the molecular structures and functionalities of lentil protein was investigated. Increasing the extraction pH to 10 caused partial protein hydrolysis and unfolding as suggested by size exclusion high performance liquid chromatography and Fourier transform infrared spectroscopy analysis, leading to improved protein solubility and gelling property. Environmental pH influenced protein solubility and surface charge, and subsequently the gelling and foaming properties. The foaming capacity was especially strong, comparable to whey and egg proteins.     

Optimisation of total phenolic acids extraction from mandarin peels using microwave energy: The importance of the Maillard reaction

Microwave processing has been used for the extraction of antioxidants from mandarin peels using deionised water as the solvent. Optimum extraction conditions under constant heat load per unit mass were as follows: microwave power, 400 W; extraction time, 3 min; solid to solvent ratio, 1:2; and extraction temperature below 135 °C. These conditions did not change the colour of the extract significantly under neutral (pH 4.5-4.9) and acidic conditions (pH 1.5-2.5). The amounts of antioxidants, which were extracted when the heat load per unit mass was held constant, were not significantly affected by the solid to solvent ratio used, so the maximum solid to solvent ratio of 1:2 used here is recommended to minimise the amount of solvent used. The colour of the extract was more greatly affected by the extraction temperature than by the solid to solvent ratio. The total phenolic contents of mandarin peels were evaluated using the Folin-Ciocalteu assay (FCR) and the trends were compared with the results of the Oxygen Absorbance Radical Capacity (ORAC) assays. The presence of Maillard rections and Maillard by-products were also investigated at higher extraction temperatures and analysed by the High Performance Liquid Chromatography (HPLC) method. The results demonstrated that microwave extraction (MWE) is a fast and reliable method for phenolic compound extraction from citrus marc with the minimum use of solvent.     

Pressurized fluid extraction of pistachio oil using a modified supercritical fluid extractor and factorial design for optimization

A pressurized fluid extraction (PFE) method for the extraction of pistachio oil was developed mainly as an analytical tool to determine oil content and/or its quality. The supercritical fluid extractor was modified to be able to pump liquid solvent and CO₂ into the extraction vessel alternatively. The extraction yield was found independent of the pressure in the range 10-150 bar tested. The addition of crushed glass increased the extraction yield by more than 15 g/100 g, while the extraction reproducibility expressed by percentage RSD was improved from 4 to 1. Furthermore, the use of crushed glass reduced the solvent consumption from 35 to 20 mL. The effective variables of temperature (40-80 °C), solvent volume (5-25 mL), and crushed glass percentage (30-60 g/100 g) were optimized by a factorial design method. The model allows the prediction of the extraction yield at different conditions. The PFE yield (i.e. 52.6 g/100 g) and fatty acid composition of pistachio oil were found similar to Soxhlet extraction and their variations were within the experimental uncertainty verified by statistical t-test analysis. Two different solvents of n-hexane and ethanol were used for PFE of pistachio oil. The extraction yield was about one-third (i.e. 18 g/100 g) when ethanol was used as solvent.     

Response surface optimisation of aqueous enzymatic oil extraction from bayberry (Myrica rubra) kernels

Aqueous enzymatic extraction (AEE) of oil from Myrica rubra kernels was performed. The four AEE parameters namely mixed enzyme (cellulose/neutral protease = 1/2, w/w) amount, liquid/solid ratio, extraction time, and temperature, were optimised by response surface methodology. The statistical analysis indicated that the enzyme amount, liquid/solid ratio, time, and the quadratics of liquid/solid ratio, and enzyme amount, as well as the interactions between time and temperature, showed significant effects on oil yield. The optimal extraction conditions for oil yield were mixed enzyme amount, liquid/solid ratio, time, temperature as 3.17% (w/w), 4.91 ml/g, 4 h and 51.6 °C. Under the optimum conditions, the experimental oil extraction yield was 31.15%. The GC–MS analysis showed that the oil was abundant in the unsaturated fatty acids (9-octadecenoic acid and 9,12-octadecadienoic acid accounting for more than 80%), and the AEE was more efficient method to extract polyunsaturated fatty acid than the organic solvent process.     

Pressurized low polarity water extraction of lignans from whole flaxseed

The application of pressurized low polarity water (PLPW) extraction to the production of flaxseed lignans-rich products has been studied, and the key geometric and process conditions, including temperature, flow rate, and total volume have been determined and optimized. Maximum amounts of lignans and other flaxseed bioactive, including proteins were extracted at 160 °C and 5.2 MPa. However, on a dry weight basis the most concentrated extracts in terms of lignans and other phenolic compounds were obtained at 140 °C and 5.2 MPa. A flow rate of 0.5 mL/min was optimal for the extraction of lignans from flaxseed (Linum usitatissimum L.) and a total volume of 30–40 mL/g of seed was required to maximize the recovery. Higher flow rates increased the rate of the extraction but required larger water volumes. Bed depth to ID ratios of 5–18 resulted in faster extraction and maximum recovery (90–95%) at water to seed ratios of 30–50 mL/g. Larger depth to ID ratios (15–18) would allow the use of lower solvent to solid ratios (14–20 mL/g) and would still result in yields of 84–90%.     

Solubility and solution thermodynamic properties of quercetin and quercetin dihydrate in subcritical water

Fundamental physicochemical data is required for the design and optimization of food engineering processes, such as extraction. Flavonoids are present in natural products such as grapes and have numerous health benefits particularly with respect to their reported antioxidant properties. Such flavonoid compounds can be extracted from these natural products using a variety of solvents, among them water. In this study, the aqueous solubilities of 3,3′,4′,5,7-pentahydroxyflavone (quercetin) and its dihydrate were measured at temperatures between 25 and 140 °C using a continuous flow type apparatus. The flow rate of subcritical water was studied at 0.1, 0.2 and 0.5 mL/min to study its effect on quercetin solubility and thermal degradation at temperatures greater than 100 °C. The aqueous solubility of anhydrous quercetin varied from 0.00215 g/L at 25 °C to 0.665 g/L at 140 °C and that of quercetin dihydrate varied from 0.00263 g/L at 25 °C to 1.49 g/L at 140 °C. The aqueous solubility of quercetin dihydrate was similar to that of anhydrous quercetin until 80 °C. At temperatures above or equal to 100 °C, the aqueous solubility of quercetin dihydrate was 1.5–2.5 times higher than that of anhydrous quercetin. The aqueous solubility of quercetin anhydrate and dihydrate at different temperatures was correlated using a modified Apelblat equation. The thermodynamic properties of the solution of quercetin and its dihydrate in water were than estimated from their solubility values. A flow rate effect on the aqueous solubility of quercetin and its dihydrate was not observed until above 100 °C where higher solvent (water) flow rates (>0.1 mL/min) were required to maintain a constant solubility in the saturation cell and with minimal thermal degradation of the solute (quercetin dihydrate). The study of its particle morphology under SEM indicated an aggregation of the crystals of quercetin dihydrate at subcritical water temperatures and at lower flow rates (<0.5 mL/min), thereby inhibiting stable solubility measurements and solvent flow through the saturation cell.     

Simultaneous determination of nine lignans using pressurized liquid extraction and HPLC-DAD in the fruits of Schisandra chinensis

The pressurized liquid extraction and HPLC-DAD was developed for extraction and determination of bioactive lignans in Schisandra chinensis. The efficient PLE conditions employed methanol as extraction solvent, 125 °C of extraction temperature, 5 min of static extraction time and only one cycle. A rapid HPLC-DAD method was described for simultaneous determination of nine lignans, including schisandrol A, gomisin J, schisandrol B, tigloylgomisin H, angeloylgomisin H, schisandrin A, γ-schisandrin, gomisin N and schisandrin C. The extraction efficiency of PLE was observed to be comparable with reflux and sonication. In addition, the contents of nine lignans in S. chinensis from different regions were analysed by PLE and HPLC-DAD method.     

Extraction conditions can greatly influence antioxidant capacity assays in plant food matrices

The estimated antioxidant capacity of different matrices can vary considerably between research reports. Besides intrinsic factors (not studied here), our work showed that may have various causes. Firstly, different methods are used to measure antioxidant capacity. Secondly, the results obtained for a single matrix by one method (such as ORAC) can vary with the extraction conditions. Parameters having a great impact on the amount and composition of antioxidants in extracts, and thus on the measured antioxidant capacity, notably include the extraction solvent composition, temperature, extraction time (duration), solvent-to-solid ratio, and storage conditions. Standardisation of the extraction procedure is thus necessary for accurate and reproducible determination of the antioxidant capacity and phenolics in different food matrices by different laboratories. In this study we optimised such a procedure for four fresh plant matrices (orange, apple, leek, and broccoli). The optimised procedure requires extraction in a mixture of acetone/water/acetic acid (70/28/2, v/v/v) for 1 h at 4 °C, with a solvent-to-solid ratio of 20 mL per 1 g. Fresh material should be used, but if this is not possible, one may lyophilise the plant matrices or store the extracts for a few days at −20 °C before analysis.     

Partial characterization of lipases produced by a newly isolated Penicillium sp. in solid state and submerged fermentation: A comparative study

The aim of this research was the partial characterization of enzymatic extracts produced by a newly isolated Penicillium sp. in submerged (SmF) and solid state fermentation (SSF). The partial characterization of the crude enzymatic extract obtained by SSF and SmF systems showed optimum activity at pH 5.5 and 47 °C, and pH 7.0 and 37 °C, respectively (15.17 U/mL and 11.28 U/mL). The crude enzymatic extracts obtained by SmF and SSF presented the best stability at pH from 4.9 to 8.5 and temperature from 25 °C to 35 °C and pH 7.0 and 25 °C, respectively. These results confirm the interesting potential of SSF, because, besides the higher activities obtained in this system, the half-life time at 25 °C was higher than that observed for the lipase extract obtained in the SmF system.     

Chemical and economic evaluation of natural antioxidant extracts obtained by ultrasound-assisted and agitated bed extraction from jussara pulp (Euterpe edulis)

This work aimed to evaluate the influence of ultrasonic and agitated bed extractions on the chemical composition and manufacturing costs of extracts obtained from jussara (Euterpe edulis) pulp. The effects of extraction time (5–180 min), temperature (25–55 °C), ethanol concentration (0–90% in acidified water) and solvent/pulp ratio (5–30 mL/g) on the extraction yield, phenolic content, anthocyanin content, antioxidant capacity and manufacturing costs were assessed. The yields provided by the ultrasound-assisted and agitated bed extractions were not significantly different. The anthocyanins and phenolic compound yields were significantly affected by the extraction time, the ethanol concentration in water and the solvent/feed ratio, but not by the temperature. In general, the antioxidant capacity of the extracts displayed tendencies similar to the anthocyanin and phenolic compound yields. The production of crude extracts obtained by ultrasound and agitated bed extraction incurred greater manufacturing costs compared to the market prices of assai extracts.     

The contents of lignans in commercial sesame oils of Taiwan and their changes during heating

Sesame lignans have multiple functions and were recently reported to have potential as sources of phytoestrogens. Sesame oils used in Taiwan are expelled from roasted sesame seeds with dark colour and strong flavour. This study analyzed lignan contents of 14 brands of sesame oils, and found their mean of total lignans to be 11.5 mg/g; 82% and 15% of the lignans were sesamin, and sesamolin, respectively. Sesamol contents were relatively higher in those with darker colour. In use as a cooking oil, heating at 180 °C for 4 min did not change the content of lignans, but the level of sesamol increased after heating at 180 °C for 20 min. Heating at 200 °C for 20 min caused a significant loss of sesamolin and sesamol. From our calculation, ingestion of 10 g of sesame oil is adequate to provide the level of lignans that might benefit cardiovascular health, as found by other studies. Cooking at temperatures above 200 °C will cause loss of some lignans, but sesamin, a source of phytoestrogen, is relatively heat-stable.     

Preparation of isoflavones enriched soy protein isolate from defatted soy hypocotyls by supercritical CO2

Soy hypocotyls, the byproduct from soy protein industry, rich in isoflavones, was attempted to develop into a value-added functional food, soy protein isolate (SPI). Soy hypocotyls were defatted by supercritical CO₂ (SC-CO₂) under the following conditions as 35 MPa, 2 h, 45 °C with a constant flow rate of CO₂ at 5 l/min, fed from the bottom of the extraction vessel. Via extraction, the oil contents were lowered from 10.5 g/100 g in original soy hypocotyls to 0.2 g/100 g in defatted soy hypocotyls (DSH) with a Protein Dispersibility Index (PDI) value of 91.2 that contained about 2.18 g/100 g of isoflavones and 63.6 g/100 g of proteins. SPI was then extracted from DSH under the optimized conditions including: the pH of water solution at 6, the ratio of DSH to water solution as 1:10 and the extraction temperature at 30 °C with the extraction time of 30 min. The freeze-dried SPI contained 92.46 g/100 g of the proteins and 640 mg/100 g of isoflavones, where the three major β-glycosides, glycitin, daidzin and genistin accounted for about 83.7% of the total isoflavones. The yields of SPI and isoflavones were about 48% and 30% in terms of their respective total contents in DSH.     

Effects of supercritical CO2 fluid parameters on chemical composition and yield of carotenoids extracted from pumpkin

Pumpkin is a traditional food that is grown extensively worldwide and is believed to be beneficial to human health due to its high contents of carotenoids. The carotenoids in pumpkin were extracted by organic solvents and by supercritical carbon dioxide (SC-CO₂), and then they were identified, quantified, and compared. β-carotene (31 to 40 g per 100 g of total carotenoids) was the predominate carotenoid in pumpkin. Lutein and lycopene contents were much higher in SC-CO₂ extracts than those in organic solvent extract. Cis-β-carotene increased by more than two times in the SC-CO₂ extracts, even at a relatively low temperature of 40 °C, over those in the solvent extracts, indicating both enhanced solubility and isomerization from trans- to cis-β-carotene. The influences of modifier (10 mL/100 mL), temperature (40-70 °C), and pressure (25-35 MPa) of SC-CO₂ extraction on the change of carotenoid yields were also investigated. The highest yield (109.6 μg/g) was obtained at 70 °C and 35 MPa, with a 73.7% recovery. Selective extraction could be achieved by adjusting the temperature and pressure. Higher proportions of all-trans-β-carotene extracts were achieved at 40 °C under both 25 MPa and 35 MPa conditions. In order to extract more cis-isomers, a higher temperature of 70 °C was preferred.     

Optimization and modeling of extraction of solid coconut waste oil

Solid coconut waste was produced after coconut milk extraction process and may still contain up to 24 wt.% oil content. In this work, extraction of oil from coconut waste using batch and soxhlet extractor was studied. Effect of particle size diameter, type of solvent and solvent to solid ratio on the kinetic and thermodynamic parameters; entropy, enthalpy and free energy of extraction were investigated. The maximum oil yields for soxhlet and batch reactor were 23.6% at 80 °C and 21.9% at 65 °C, respectively for particle size diameter <0.5 mm when hexane was used as solvent. The kinetic of coconut waste oil extraction was found to be a first order mass transfer model. The ΔG, ΔS and ΔH values were 10.94–13.35 kJ/mol, 33.10–39.57 J/mol K and 0.12–1.25 kJ/mol, respectively shows that the extraction process was spontaneous, irreversible and endothermic based on thermodynamic parameters.     

Ultrasound-assisted solid liquid extraction (USLE) of olive fruit (Olea europaea) phenolic compounds

A new method of ultrasound-assisted solid liquid extraction (USLE) of olive fruit phenols is described. Phenolics were extracted using high intensity probe ultrasonication and analysed by HPLC-DAD-FLD-MS/MS. Four USLE parameters – sonication time (4, 15, 20, 30 min), temperature (25, 45 °C), solvent composition (80%, 100% methanol) and extraction steps (1–5) were studied and optimised on the basis of nine major olive fruit phenols. A three-step extraction of 20 min with pure methanol (25 mL) at 45 °C was needed for sufficient phenol recoveries (94.1–98.7%) from 1.5 g of freeze-dried olive fruits. The proposed USLE method was more efficient in comparison to US bath and agitation, with up to 33% and 80% enhancement in the case of oleuropein, respectively. In addition, the overall method provided high selectivity, precision and sensitivity with LODs/LOQs ranging from 0.66–4.92 μg g⁻¹ and 2.00–14.77 μg g⁻¹ of olives DW, respectively.