Optimization of an Enzyme-Assisted Extraction Method for the Anthocyanins Present in Açai (Euterpe oleracea Mart.)

Several investigations have proven the presence of anthocyanins in different parts of acai plants. These compounds are responsible for the notable therapeutic properties of acai such as antioxidant, antimicrobial, anti-inflammatory, anticancer, and anticonvulsant. We have therefore optimized an enzyme-assisted extraction method for the anthocyanins found in acai, to be subsequently applied in many fields such as agrifood, medicine, or cosmetics. A Plackett-Burman design with seven variables (time of extraction, pH, temperature, agitation, percentage of ethanol in the solvent, amount of sample, and units of enzyme) was employed to determine the predominant extraction variables, of which four were categorized as influential. Subsequently, a Box-Behnken design-response surface methodology made it possible to determine the degree of influence from these variables and their optimal values. The optimal conditions were established as 0.1 g of acai heated up to 60 degrees C and extracted using 15 mL of solvent with pH 4 and 40% ethanol, 500 units of enzyme per gram of sample, and agitation at 150 rpm for 15 min. The repeatability and intermediate precision of the developed method were confirmed by variation coefficients below 5%. Finally, the developed method was compared against the extensively used maceration and ultrasound-assisted extraction methods.info:eu-repo/semantics/publishedVersio

Enhancing efficiency of enzymatic-assisted extraction method for evaluating bioactive compound analysis in mulberry: an optimization approach

The present investigation endeavors to optimize a method based on enzyme-assisted extraction for the efficient retrieval of bioactive compounds from mulberry, leveraging its notable health-promoting properties. A combined approach of Plackett-Burman design followed by Box-Behnken design was employed for determining the crucial extraction parameters and subsequently, refining the process. Optimal conditions consisted of heating 0.15 g of mulberry at 40 degrees C, using 15 mL of 70% EtOH as a solvent at pH 4, 38.46 enzyme units per g of sample, and shaking at 200 rpm. The optimum extraction time study revealed that 5 min of extraction was sufficient to reach the maximum concentration of the bioactive compound. The repeatability and intermediate precision assessment exhibited a coefficient of variation below 5%. Among the diverse mulberry varieties scrutinized, Morus nigra showed the highest anthocyanin content (27.90 +/- 2.14 mg/100 g), while Morus rubra showed the highest concentration of phenolic compounds (121.10 +/- 19.56 mg/100 g). Moreover, the extracted compounds showcased significant antioxidant and antimicrobial properties.Equipment for liquid chromatography by means of mass spectrometry and ion chromatography) of the State Subprogram of Research Infrastructures and Technical Scientific Equipment EQC2018-005135-Pinfo:eu-repo/semantics/publishedVersio

A comparison study between ultrasound-assisted and enzyme-assisted extraction of anthocyanins from blackcurrant (Ribes nigrum L.)

Blackcurrant (Ribes nigrum L.) is a fruit rich in vitamins, fatty acids, minerals, essential oils and phenolic com-pounds, including anthocyanins. In the present work, two anthocyanin extraction methods from blackcurrant samples based on Ultrasound-Assisted Extraction (UAE) and Enzyme-Assisted Extraction (EAE) have been developed. A Plackett-Burman design with seven variables has been preliminary used for both UAE and EAE in order to determine the most influential variables in each methodology. After that, a Box-Behnken design was employed to optimize the extraction methods. The composition of the extraction solvent (% EtOH in water) has been the most influential variable for both UAE and EAE. The optimal extraction times have been 5 min for UAE and 10 min for EAE. No differences have been observed in anthocyanin extraction with both methodologies. Both methods have been applied to blackcurrant-derived products and proven their suitability for quality control analysis.This work has been supported by the project "EQC2018-005135-P" (Equipment for liquid chromatography by means of mass spectrometry and ion chromatography) of the State Sub-program of Research In-frastructures and Technical Scientific Equipment

Extraction of Anthocyanins and Total Phenolic Compounds from Açai (Euterpe oleracea Mart.) Using an Experimental Design Methodology. Part 3: Microwave-Assisted Extraction

In this work, two methods based on microwave-assisted extraction techniques for the extraction of both anthocyanins and total phenolic compounds from acai have been developed. For that, a full factorial design (Box-Behnken design) has been used to optimize the following four variables: solvent composition (25-75% methanol in water), temperature (50-100 degrees C), pH (2-7), and sample/solvent ratio (0.5 g: 10 mL-0.5 g: 20 mL). The anthocyanins and total phenolic compounds content have been determined by ultra high-pressure liquid chromatography and Folin-Ciocalteu method, respectively. The optimum conditions for the extraction of anthocyanins were 38% MeOH in water, 99.63 degrees C, pH 3.00, at 0.5 g: 10 mL of ratio, while for the extraction of total phenolic compounds they were 74.16% MeOH in water, 99.14 degrees C, pH 5.46, at 0.5 g: 20 mL of ratio. Both methods have shown a high repeatability and intermediate precision with a relative standard deviation lower than 5%. Furthermore, an extraction kinetics study was carried out using extraction periods ranging from 2 min until 25 min. The optimized methods have been applied to acai-containing real samples. The results with such real samples have confirmed that both methods are suitable for a rapid and reliable extraction of anthocyanins and total phenolic compounds

Extraction of anthocyanins and total phenolic compounds from açai (euterpe oleracea mart.) using an experimental design methodology. part 1: Pressurized liquid extraction

Currently, açai is one of the most important fruits present in the world. Several studies have demonstrated its high content in phenolic compounds and anthocyanins. Both of them are responsible of interesting properties of the fruit such as anti-inflammatory, antioxidant or anticancer. In the present study, two optimized pressurized liquid extraction (PLE) methods have been developed for the extraction of anthocyanins and total phenolic compounds from açai. A full factorial design (Box-Behnken design) with six variables (solvent composition (25%-75% methanol-in-water), temperature (50-100°C), pressure (100-200 atm), purge time (30-90 s), pH (2-7) and flushing (50%-150%)) were employed. The percentage of methanol in the extraction solvent was proven to be the most significant variable for the extraction of anthocyanins. In the case of total phenolic compounds, the extraction temperature was the most influential variable. The developed methods showed high precision, with relative standard deviations (RSD) of less than 5%. The applicability of the methods was successfully evaluated in real samples. In conclusion, two rapid and reliable PLE extraction methods to be used for laboratories and industries to determine anthocyanins and total phenolic compounds in açai and its derived products were developed in this work

Extraction of Anthocyanins and Total Phenolic Compounds from Acai (Euterpe oleracea Mart.) Using an Experimental Design Methodology. Part 2: Ultrasound-Assisted Extraction

Two optimized methods for ultrasound-assisted extraction were evaluated for the extraction of two types of acai bioactive compounds: Total anthocyanins (TAs) and total phenolic compounds (TPCs). For the extraction optimization, a Box Behnken factorial design of different variables in the following intervals was used: Methanol-water (25%-75%) for solvent composition, temperatures between 10 and 70 degrees C, amplitude in the range between 30% and 70% of the maximum amplitude -200 W), extraction solvent pH (2-7), the ratio for sample-solvent (0.5 g:10 mL-0.5 g:20 mL), and cycle between 0.2 and 0.7 s. The extraction kinetics were studied using different periods between 5 and 30 min. TA and TPC were analyzed by UHPLC and the Folin-Ciocalteu method, respectively. Optimized conditions for TA were: 51% MeOH in water, 31 degrees C temperature, pH 6.38, cycle 0.7 s, 65% amplitude, and 0.5 g:10 mL of sample-solvent ratio. Optimized conditions for the TPC were: 49% MeOH in water, 41 degrees C temperature, pH 6.98, cycle 0.2 s, 30% amplitude, and 0.5 g:10 mL of sample-solvent ratio. Both methods presented a relative standard deviation below 5% in the precision study. The suitability of the methods was tested in real samples. It was confirmed that these methods are feasible for the extraction of the studied bioactive compounds from different acai matrices

Ultrasound-assisted extraction of betalains from opuntia fruit pulp of different color varieties

Betalains are water-soluble pigments that have exhibited important pharmacological properties such as antioxidant, anticancer, antilipidemic and antimicrobial activity. These compounds have been isolated in numerous purple plants or fruits, as is the case of the wild species under the Opuntia genus. The fruits of these species are often disregarded because of their small size as well as the frequent presence of prickles. Based on this, this research has as its objective the optimization of a method based on ultrasound-assisted extraction to obtain extracts enriched with betalains from a wild Opuntia species (Opuntia dillenii (Ker Gawl.) Haw.). Four variables (%EtOH in the solvent, temperature of extraction, ultrasound amplitude and cycle) were selected using a Box-Behnken design. The quadratic interaction of %EtOH and the interaction of %EtOH-cycle have proven to be influential variables at 95% confidence. The conditions to obtain the highest betalain concentration were 100 mg of pulp with 20 mL (60%:40% EtOH:H2O) solvent at 20 degrees C at 24% amplitude and 0.2 cycle for 10 min. The suitability and reliability of the method were evaluated with repeatability and intermediate precision tests obtaining CVs <5%. Finally, the developed method has been employed in the analysis of five Opuntia commercial samples and obtained significant antioxidant activity of the extracts, confirming its applicability.info:eu-repo/semantics/publishedVersio

UV-Vis Spectrophotometry and UPLC-PDA Combined with Multivariate Calibration for Kappaphycus alvarezii (Doty) Doty ex Silva Standardization Based on Phenolic Compounds

The algae Kappaphycus alvarezii is considered an important raw material for industrial practices, producing high economic value of various derived products. However, the quality of this commodity, which can be indicated by the level of phenolic compounds, may vary due to growth factors, including cultivation sites. An analytical UV-Vis spectrophotometry method coupled with chemometrics was proposed to standardize the red alga based on the content of phenolic compounds. The correlation between the UV-Vis spectra and UPLC-PDA results, combined with a multivariate calibration of the K. alvarezii extracts, was analyzed. The extracts were prepared using an ultrasound-based technique and subsequently subjected to UV-Vis spectral measurements at 200-800 nm and UPLC-PDA at 260 and 330 nm. Chemometric techniques and partial least squares (PLS) were applied to the acquired data to build a reliable analysis of the phenolics in the K. alvarezii extracts. The result showed that the wavelength combination of 200-450 and 600-690 nm provided a valid method for quantitative analysis of the studied phenolics that belong to hydroxybenzoic acid, hydroxycinnamic acid, and flavonoid with a coefficient of regression (R-2) > 0.96 in the calibration and validation models, along with an RMSEC and RMSEP value < 8%. The method was then employed to characterize the K. alvarezii samples from 13 different cultivation areas. Principal component analysis (PCA) generated principal components that produced a clear distribution among the samples of K. alvarezii based on phenolic compounds corresponding to the geographical origin.This research forms part of the National Research Priority (PRN) project on the standardization of Indonesian algae funded by the BOPTN Research Grant 006/E4.1/AK.04.PRN/2021 from the Directorate General of Higher Education, Ministry of Education and Culture, Republic of Indonesia. The APC was supported by Universitas Gadjah Mada through the Final Project Recognition Program (Rekognisi Tugas Akhir, RTA) 2020 with assignment number 3143/UN1.P.III/DIT-LIT/PT/2021

Optimization by Means of Chemometric Tools of an Ultrasound-Assisted Method for the Extraction of Betacyanins from Red Dragon Fruit (Hylocereus polyrhizus)

Betacyanins have been reported to provide prominent health-promoting effects, in addition to contributing to the exotic color and pleasant palate of red dragon fruit that has been attracting universal interest for over a decade. An accurate determination of betacyanins in dragon fruit is important if we are to benefit from the nutraceutical features of these compounds. Seven betacyanin pigments have been identified and extracted by means of ultrasound-assisted techniques from red dragon fruit. Chemometric tools such as Box-Behnken Design (BBD) in conjunction with response surface methodology (RSM) have been successfully used to evaluate and optimize the relevant extraction variables i.e., temperature (A: 10, 35, 60 degrees C), solvent composition (B: 20, 50, 80% methanol in water), sample to solvent ratio (C: 0.1:10, 0.2:10, 0.3:10 g/mL), power (D: 20, 45, 70%), and cycle (E: 0.3, 0.5, 0.7 s(-1)). The results obtained from the analysis of variance (ANOVA) suggest that the solvent composition (p 0.0063), sample to solvent ratio (p 0.0126), and cycle (p 0.0302) are the most influential variables in betacyanin extraction. The optimal variable settings for ultrasound-assisted extraction (UAE) were a 54.6 degrees C extraction temperature, 0.3 s(-1) cycle, 20% ultrasound power, 21.4% methanol in water solvent composition, 0.2:10 sample to solvent ratio, and a 5 min extraction time. The validation of the method for repeatability and intermediate precision provided excellent results at 1.56% and 2.94% respectively. Furthermore, the novel UAE method was successfully used for the determination of betacyanins content in red dragon fruit from different geographic origins.This work has been supported by the project "EQC2018-005135-P" (Equipment for liquid chromatography by means of mass spectrometry and ion chromatography) of the State Subprogram of Research Infrastructures and Technical Scientific Equipment

How Different Cooking Methods Affect the Phenolic Composition of Sweet Potato for Human Consumption (Ipomea batata (L.) Lam)

In recent years, there has been increasing interest in the functional components of sweet potato because of its nutritional and medicinal value. The aim of this study is to analyse how much sweet potato phenolic compounds composition (derived from caffeoylquinic acids) varies as a result of cooking. Traditional techniques such as: boiling, oven roasting and more recent processing techniques such as microwave cooking were tested. Three sweet potato varieties were cooked for different periods of time and under different conditions. Ultrasound-assisted extraction (UAE) was used to extract the compounds of interest and then, a chemometric tool such as Box-Behnken design (BBD) was successfully used to evaluate and optimise the most influential factors in the extraction, i.e., temperature, solvent composition and sample-to-solvent ratio. The optimal settings for UAE were: solvent 100% methanol, a temperature of 39.4 degrees C and a mass/volume ratio of 0.5 g per 10 mL solvent. Oven roasting of sweet potatoes resulted in increased levels of caffeoylquinic acids, whereas prolonged cooking times in water resulted in decreasing levels of the same.This work has been supported by the project "EQC2018-005135-P" (Equipment for liquid chromatography by means of mass spectrometry and ion chromatography) of the State Subprogram of Research Infrastructures and Technical Scientific Equipment