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.     

Optimisation of the solvent extraction of bioactive compounds from Parkia speciosa pod using response surface methodology

Central composite design was employed to optimise the buffer-to-solids ratio (X₁: 20–50 ml/g), incubation temperature (X₂: 35–55 °C) and time (X₃: 100–200 min), obtaining extracts from Parkia speciosa pod with high total phenolic and flavonoid contents and high antioxidant activities. Analysis of variance showed that the contribution of a quadratic model was significant for the responses. An optimisation study using response surface methodology was performed and 3D response surfaces were plotted from the mathematical models. The optimal conditions based on combination responses were: X₁ = 20 ml/g, X₂ = 35–36 °C and X₃ = 100–102 min. These optimum conditions yielded total phenolic contents of 664–668 mg gallic acid equivalents/100 g, total flavonoid contents of 47.4–49.6 mg pyrocatechol equivalents/100 g, %DPPH_sc of 81.2–82.1%, %ABTS_sc of 78.2–79.8% and FRAP values of 3.2–3.3 mM. Close agreement between experimental and predicted values was found. This methodology could be applied in the extraction of bioactive compounds in the natural product industry.     

Optimizing conditions for anthocyanins extraction from purple sweet potato using response surface methodology (RSM)

Optimization for purple sweet potato (Ipomoea batatas (L.) Lam) anthocyanins (PSPAs) extraction was investigated using response surface methodology in this paper. PSPAs were extracted using acid-ethanol at different extraction temperature (40-80 °C), time (60-120 min) and solid-liquid ratio (1:15-1:30). The combined effects of extraction conditions on PSPAs yield and color attributes (expressed as L*, C* and H) were studied using a three-level three-factor Box-Behnken design. The results showed that The highest yield (158 mg/100 g dw) of PSPAs were reached at the temperature 80 °C, extraction time 60 min, and solid-liquid ratio 1:32. PSPAs yield indicated a high and significant correlation with L^* (P<0.05; r=−0.961) and it was significantly affected by extraction temperature (P<0.01) and solid-liquid ratio (P<0.05).     

Optimization of the extraction of flavanols and anthocyanins from the fruit pulp of Euterpe edulis using the response surface methodology

The response surface methodology (RSM) was employed to optimize the ultrasound extraction of flavanols and anthocyanins from the pulp of jussara (Euterpe edulis), using a second-order polynomial equation to describe the experimental data for total flavanol (TF), total phenolic (TP), and total monomeric anthocyanin (TMA) contents, as well as the total antioxidant activity (TAA). A central composite design with two-variables (extraction time and solid to liquid ratio) was then applied. The optimized conditions that maximized the yields of flavanol-enriched extract were a solvent methanol/0.1 M HCl, solid to liquid ratio of between 1:50 and 1:100 and extraction time of 15 min. For anthocyanin-enriched extracts the respective optimal parameters were a solvent methanol/1.5 M HCl, solid to liquid ratio of between 1:30 and 1:50 and extraction time of 24 h. The results showed good fits with the proposed model for both the flavanol-enriched extract (R² = 0.94) and for the anthocyanin-enriched extract (R² = 0.99).     

Optimization of nutrient-induced germination of Bacillus sporothermodurans spores using response surface methodology

Spores of Bacillus sporothermodurans are known to be contaminant of dairy products and to be extremely heat-resistant. The induction of endospore germination before a heat treatment could be an efficient method to inactivate these bacteria and ensure milk stability. In this study, the nutrient-induced germination of B. sporothermodurans LTIS27 spores was studied. Testing the effect of 23 nutrient elements to trigger an important germination rate of B. sporothermodurans spores, only d-glucose, l-alanine, and inosine were considered as strong independent germinants. Both inosine and l-alanine play major roles as co-germinants with several other amino acids. A central composite experimental design with three factors (l-alanine, d-glucose, and temperature) using response surface methodology was used to optimize the nutrient-induced germination. The optimal rate of nutrient-induced germination (100%) of B. sporothermodurans spores was obtained after incubation of spore for 60 min at 35 °C in presence of 9 and 60 mM of d-glucose and l-alanine, respectively. The results in this study can help to predict the effect of environmental factors and nutrients on spore germination, which will be beneficial for screening of B. sporothermodurans in milk after induction their germination. Moreover, the chosen method of optimization of the nutrient-induced germination was efficient in finding the optimum values of three factors.     

Optimization of subcritical fluid extraction of seed oil from Nitraria tangutorum using response surface methodology

Subcritical fluid extraction (SFE) technology was used to extract oil from Nitraria tangutorum seed. The best possible combination of extraction parameters was found using response surface methodology (RSM) in a three-variable, three-level Box-Behnken experimental design (BBD). The optimum extraction parameters were an extraction time of 40 min, an extraction pressure of 0.60 MPa, an extraction temperature of 44 °C and a raw material particle size of 0.45 mm. Conventional solvent extraction and supercritical CO₂ fluid extraction were comparatively used. The yield of seed oil obtained using SFE was 12.92%, which was similar to or higher than the other methods. The chemical compositions of the seed oil, determined by GC–MS, indicate that its unsaturated fatty acids content was 97%. SFE proved to be an effective technique for extracting oil from N. tangutorum.     

Optimization of ionic liquid based ultrasonic assisted extraction of puerarin from Radix Puerariae Lobatae by response surface methodology

Ionic liquid (IL) based ultrasonic assisted extraction (ILUAE) was developed for the effective extraction of puerarin from Radix Puerariae Lobatae (RPL). The ILUAE parameters including the type of ILs, IL concentration, RPL amount, ultrasonic power and time were optimized by single-factor experiment and response surface methodology. Under the optimized experimental conditions, the best results were obtained using RPL amount 0.43 g in 10 mL 1.06 mol L⁻¹ 1-butyl-3-methylimidazolium bromide aqueous solution, ultrasonic time 27.43 min and ultrasonic power 480 W. Scanning electron microscope images of RPL samples were obtained to provide visual evidence of the sonication effect. Compared with the conventional ultrasonic assisted extraction and refluent extraction, the proposed ILUAE offered shorter extraction time and remarkable higher efficiencies due to the higher penetration ability and solubility of IL and the cavitation phenomenon produced in the solvent by the passage of an ultrasonic wave, which further supported the suitability of the proposed approach.     

Effect of thermal treatments on functional properties of cress seed (Lepidium sativum) and xanthan gums: A comparative study

Foods are generally subjected to thermal treatments during processing operations, which affect on the functional properties of hydrocolloid solutions. In this paper, the effect of different thermal conditions (60 °C-30 min, 80 °C-23 min, 100 °C-18 min and 121 °C-15 min) on functional properties (rheological, emulsifying and foaming properties) of cress seed gum and xanthan gum were investigated. The results demonstrated that cress seed gum solutions had desirable rheological, emulsifying and foaming properties. When the hydrocolloid samples were heated, an irreversible increase in viscosity of cress seed gum solutions was observed and as a result, the emulsification and foaming properties improved. Whereas, xanthan gum would not have the ability to stand against heat treatment and its viscosity decreased. Nevertheless, the viscosity values of xanthan gum solutions were always higher than cress seed gum in all conditions.     

Extraction optimization of bioactive compounds (crocin, geniposide and total phenolic compounds) from Gardenia (Gardenia jasminoides Ellis) fruits with response surface methodology

Response surface methodology (RSM) was employed to optimize the extraction parameters of crocin, geniposide and total phenolic compounds from gardenia fruits. The effects of three independent variables, namely ethanol concentration (EtOH, %), extraction temperature (°C) and time (min) on the yield of crocin, geniposide and total phenolic compounds were investigated. Results indicated that the data were adequately fitted into three second-order polynomial models. The independent variables, the linearity of extraction temperature and time, the quadratics of ethanol concentration and extraction time, the interactions between ethanol concentration and temperature, ethanol concentration and extraction time, as well as extraction temperature and time had a significant effect on the yield of crocin, geniposide or total phenolic compounds. The optimal extraction parameters were the EtOH of 51.3%, extraction temperature of 70.4 °C and time of 28.6 min according to the response surface analysis. Under this condition, the yield of crocin, geniposide and total phenolic compounds was 8.41 mg/g, 109.0 mg/g and 24.97 mg CAE/g dry powder of gardenia fruits, respectively.

Industrial relevance

There is a strong interest in the food industry in studying the extraction optimization of natural plants to produce higher quality products. Response surface methodology (RSM) is an effective technique for analyzing interactions among factors and optimizing the processes or products where multiple variables may influence the outputs. This work explores the extraction conditions for a natural pigment-gardenia yellow pigment by the RSM method and optimizes the extraction parameters to improve extraction yields of same main components of the natural pigment at the same time. The mathematical methods and models which can describe and predicate experimental data of the extraction would be extremely helpful in the extraction process of the natural products.     

Supercritical CO2 extraction optimization of pomegranate (Punica granatum L.) seed oil using response surface methodology

This study examined extraction of pomegranate seed oil by using supercritical carbon dioxide. Response surface methodology was used to evaluate the effects of the process parameters, namely extraction pressure, temperature and CO₂ flow rate on the yield of pomegranate seed oil. The extraction parameters were optimized with a central composite design experiment. The linear term of pressure, followed by the linear term of CO₂ flow rate, the quadratic terms of pressure, temperature and CO₂ flow rate and the interactions between pressure and temperature, as well as CO₂ flow rate and temperature, had significant effects on the oil yield (p < 0.05). Maximum yield of pomegranate seed oil from the mathematical model was predicted to be 156.3 g/kg dry basis under the condition of pressure 37.9 MPa and temperature 47.0 °C with CO₂ flow rate of 21.3 L/h. The fatty acid composition and the tocopherols' content of pomegranate seed oil extracted using supercritical CO₂ were compared with those obtained by Soxhlet method. Minor difference was found in the fatty acid composition of the oils extracted by the two methods. The content of total tocopherols was about 14% higher in the oil extracted with supercritical CO₂ than that obtained by Soxhlet extraction.     

Response surface methodology for optimization of extraction yield, viscosity, hue and emulsion stability of mucilage extracted from Lepidium perfoliatum seeds

Response surface methodology was used to determine the optimum processing conditions that give maximum extraction yield, viscosity, hue and emulsion stability, as well as, minimum protein content for the gum extracted from Lepidium perfoliatum seed. Temperature (45–75 °C), processing time (1.5–3.5h), pH (5–8) and water to seed ratio (30:1–60:1) were the factors investigated. Experiments were designed according to Central Composite Rotatable Design with these four factors, including central and axial points. For each response, a second-order polynomial model was developed using multiple linear regression analysis. Applying desirability function method, optimum operating conditions were found to be extraction temperature of 48.1 °C, pH of 8, water to seed ratio of 30:1 and process time of 1.5 h. At this optimum point, extraction yield, viscosity, protein content, hue and emulsion stability were found to be 17.36%, 463.07 mPa s, 2.84%, 60.47 and 88.96 %, respectively.     

Rheological properties of Lepidium sativum seed extract as a function of concentration, temperature and time

The seeds of Lepidium sativum (Garden Cress) were selected as a new source of hydrocolloid and its chemical composition and molecular parameters were determined. The macromolecular component of the extract had a molecular weight of 540 kDa, and was nearly as rigid as xanthan with regard to chain conformation. The main rheological features were investigated as a function of shear rate, concentration and temperature. The extract exhibited strong shear-thinning behaviour, which was even more pronounced than for xanthan. An increase in concentration or temperature led to an increase in pseudoplasticity. The Arrhenius model was applied to the temperature dependence of viscosity, and the activation energy (E_a) was found to decrease with increasing concentration. The extract solutions showed thixotropic behaviour at all the concentrations and temperatures studied, and the first-order stress decay model with a non-zero equilibrium stress fairly described the time-dependent behaviour. The rheological characteristics found indicated a potential application of the extract as a novel thickener.     

Production of a diacylglycerol-enriched palm olein using lipase-catalyzed partial hydrolysis: Optimization using response surface methodology

Partial hydrolysis using Lipozyme RMIM lipase in a solvent-free system was used to produce a diacylglycerol (DAG)-enriched palm olein. Response surface methodology (RSM) was applied to model and optimize the reaction conditions namely water content (30–70 wt% of enzyme mass), enzyme load (5–15 wt% of oil mass), reaction temperature (45–85 °C) and reaction time (6–16 h). Well fitting models were successfully established for both DAG yield (R² = 0.8788) and unhydrolysed triacylglycerol (TAG) (R² = 0.8653) through multiple linear regressions with backward elimination. Chi-square test indicated that there were no significant (P > 0.05) differences between the observed and predicted values for both models. All reaction conditions had positive effects on DAG yield and negative effects on unhydrolysed TAG. Optimal reaction conditions were: 50 wt% water content, 10 wt% enzyme load, 65°C of reaction temperature and 12 h of reaction time. The process was further up-scaled to a 9 kg production in a continuous packed bed bioreactor. Results indicated that upscaling was possible with a similar DAG yield (32 wt%) as in lab scale. Purification of the DAG oil using short path distillation yielded a DAG-enriched palm olein with 60 wt% DAG and 40 wt% TAG which is suitable for margarine, spread or shortening applications.     

Optimization of medium composition for enhanced chitin extraction from Parapenaeus longirostris by Lactobacillus helveticus using response surface methodology

Chitin extraction by biological way, using the lactobacilli Lactobacillus helveticus, is a non-polluting method and offers the opportunity to preserve the exceptional qualities of chitin and its derivatives. However, the major disadvantage of the fermentative way is the low efficiency of demineralization and deproteinization. The aim of our study is to improve the yield of extraction.     

Antimicrobial activity of essential oil extracts of various onions (Allium cepa) and garlic (Allium sativum)

Antimicrobial activity of different concentrations (50, 100, 200, 300 and 500 ml/l) of essential oil extracts of three type of onions (green, yellow and red) and garlic against two bacteria, Staphylococcus aureus, Salmomella Enteritidis, and three fungi, Aspergillus niger, Penicillium cyclopium and Fusarium oxysporum, was investigated. The essential oil (EO) extracts of these Allium plants (garlic and onions) exhibited marked antibacterial activity, with garlic showing the highest inhibition and green onion the lowest. Comparatively, 50 and 100 ml/l concentrations of onions extracts were less inhibitory than 200, 300 and 500 ml/l concentrations. However, with garlic extract, high inhibitory activity was observed for all tested concentrations. S. aureus showed less sensitivity towards EO extracts inhibition, however S. Enteritidis was strongly inhibited by red onion and garlic extracts. The fungus F. oxysporum showed the lowest sensitivity towards EO extracts, whereas A. niger and P. cyclopium were significantly inhibited particularly at low concentrations. Conclusively, where seasoning is desired, essential oil extracts of onions and garlic can be used as natural antimicrobial additives for incorporating in various food products.     

Characterisation of lipoxygenase from pea seeds (Pisum sativum var. Telephone L.)

Lipoxygenase (LOX) from pea seeds (Pisum sativum var. Telephone L.) was extracted and studied of biochemical properties. The molecular mass of purified lipoxygenase was 93 kDa. The effects of substrate specificity, pH, and sensibility to various inhibitors: caffeic acid, ferulic acid, benzoic acid, catechin, quercetin and kaempferol of LOX were investigated. Lipoxygenase showed the highest activity toward linoleic acid and the lowest toward oleic acid as substrates. Kinetic studies indicated that V_max of the LOX activity was 151.5 U/min and corresponding K_m value of 0.44 × 10⁻³ M. Optimum pH of lipoxygenase was reported at 5.5. Caffeic acid was the most effective inhibitor and kaempferol was the least effective.     

Response surface optimised extraction and chromatographic purification of rosmarinic acid from Melissa officinalis leaves

The extraction of lemon balm (Melissa officinalis) leaves with aqueous methanol was optimised using response surface methodology. Fifteen runs were conducted following a Box-Behnken design (BBD) followed by ridge analysis using the concentration of methanol, the extraction temperature and time as the independent variables and taking the extraction yield of RA from lemon balm as the response variable. The optimal extraction conditions were a methanol concentration of 59.0% (v/v), a temperature of 54.8 °C and a time of 64.8 min, which gave a maximal RA yield of 46.1 mg RA/g dry materials. The RA extract was loaded onto a column packed with Sephadex LH-20 and then was eluted with 100% methanol, which resulted in RA with a purity of 38.8% and a yield of 43.8%. The purity of RA increased by 3.1-fold when compared to its initial purity in the extract obtained from extraction.     

Production of theanine by Xerocomus badius (mushroom) using submerged fermentation

Theanine is a rare amino acid mainly produced by the genus Camellia, which is also found in the Basidiomycete fungus (mushroom) Xerocomus badius. Besides its favorable taste, broad potential physiological effects have been reported in the recent years, leading to the very fast growing demand for theanine worldwide. Thus a series of methods to produce theanine have been developed, including tea callus cultivation, chemical synthesis, extraction from the fresh leaves of the tea plant, and enzymatic synthesis. Different from the previous procedures, we reported here a novel protocol to produce theanine from the mushroom X. badius by means of submerged fermentation. Moreover, response surface methodology (RSM) was employed to optimize various parameters during fermentation. The statistic results indicated that the maximum theanine yield of X. badius (17.2420 mg/L) could be achieved at the optimal conditions, with glucose of 29.17 g/L, yeast extract of 6.38 g/L in a broth volume of 61.43 mL (within a flask of 250 mL) at an initial pH at 5.63. These predicted values were also validated by experiments. Taken together, these findings provided us a promising strategy for the production of theanine from X. badius in addition to the traditional preparation processes.     

Optimization of extraction process of crude polysaccharides from boat-fruited sterculia seeds by response surface methodology

Response surface methodology (RSM) was applied to optimize the extraction of crude polysaccharides from boat-fruited sterculia seeds. A central composite design was used for experimental design and analysis of the results to obtain the optimal extraction conditions. Extraction temperature, pH, extraction time and water to seed ratio were found to have a significant influence on the yield and purity of the extracted crude polysaccharides, while the three other factors except the water to seed ratio also significantly affected the relative viscosity. Based on the RSM analysis, optimum conditions were: temperature 60–65 °C, time 2.3–3.1 h, pH at 7.0 and water to seed ratio at 75:1. Under the optimized conditions, the experimental values were in close agreement with values predicted by the model. The crude polysaccharides prepared under optimum conditions contained 58.2% total carbohydrates (including uronic acids), 20% proteins, 9% moisture and 4.5% ash. The crude polysaccharides consisted of glucose (22.6%), rhamnose (10.0%), arabinose (7.9%), galactose (5.0%), xylose (0.8%) and galacturonic acid (11.8%).     

Formulation of pasta from rice brokens: optimization of ingredient levels using response surface methodology

Rice brokens were utilized in the development of pasta products. Response surface methodology was used to analyze the effect of pregelatinized rice flour (from rice brokens), vital gluten, water, glycerol monostearate (GMS) and carboxy methyl cellulose (CMC) on the quality responses (sensory, cooking quality, rehydration ratio and solids loss) of the pasta product. A rotatable central-composite design was used to develop models for the responses. Responses were affected most by changes in rice flour and vital gluten levels and to a lesser extent by water, GMS and CMC levels. The maximum sensory score, cooking quality, rehydration ratio and minimum solids loss were identified at 682.54 g/kg rice flour, 235.85 g/kg water, 70.70 g/kg vital gluten, 1.59 g/kg GMS and 9.33 g/kg CMC level.