Ultrasound-Assisted Extraction of L-Tryptophan from Chamomile Flower: Method Development and Application for Flower Parts Characterization and Varietal Difference

Journal Article
Oktaviani NMD, Larasati ID, Nugroho AW, Setyaningsih W, Palma M

Abstract: Chamomile has been widely used as a functional tea due to its effects on several neurohormones, some of which are related to tryptophan. An analytical ultrasound-assisted extraction (UAE) technique was successfully optimized and validated to determine tryptophan levels in chamomile flowers. A Box-Behnken design,in conjunction with response surface methodology,was conducted based on 3factors and 3levels: Temperature (x1; 30, 50, and 70 °C), solvent composition (x2; 0, 40, and 80% methanol in water), and ultrasonic power (x3; 20, 60, and 100%). The main (x2) and quadratic effect of solvent (x2x2); quadratic of temperature (x1x1) and ultrasonic power (x3x3); also, the interaction of solvent-ultrasonic power (x2x3) and temperature-ultrasonic power (x1x3), significantly affected (p<0.05) the level of L-tryptophan in the extracts. Optimum extraction conditions were achieved by applying a temperature of 54 °C for 15 min using 17% methanol in water and 69% ultrasonic power. A high recovery (94.26%) was achieved with2extraction cycles. The coefficient of variation (CV) demonstrated that the developed method had a satisfactory level of precision (CV<5%) for repeatability and intermediate precision. To check the applicability of the method, the parts of 2types of chamomile flowers (German and Roman) were evaluated. This method was successfully applied to characterize chamomile flowers based on L-tryptophan levels.

Incorporation of Young Coconut (Cocos nucifera)Mesocarp Increases the Antioxidant Activity, Phenolic Compounds and Oxidative Stability of Cookies

Journal Article
Leliana L, Setyaningsih W, Palma M, Supriyadi, Santoso U

Abstract: Young coconut mesocarp (YCM) contains considerable phenolic compounds that provide high antioxidant activity. However, the YCM is identified as an underutilized by-product in food industries. A novel functional food derived from YCM, viz., cookies, was proposed through this study to take advantage of the containing antioxidants. The cookies were prepared using regular flour (CR) or substituted with YCM powder at 8 (C8) or 16 % (C16). The macro components (carbohydrate, protein, fat and ash), in vitroantioxidant activities, and the level of individual phenolic compounds in the cookies were measured. The addition of YCM powder increased the crude fiber by up to 5 % and moisture content by about 2 – 3 %. Conversely, the fat level was lowered by 1 %. The in vitro antioxidant activities of cookies C8 and C16 were noticeably increased compared to the CR. The increase was attributable to the presence of phenolic compounds such as catechin and 3 caffeic acid derivatives, which were retained after the baking process. The addition of YCM changed the visual appearance by darkening the color of cookies which was indicated by lightness at 45.33 and 23.87; a* value was 17.30 and 18.97; b* value was 29.47 and 19.83; and chroma at 34.15 and 27.49, for C8 and C16, respectively. The addition of YCM also reduced the hardness due to the presence of moisture content in the incorporated cookies. Additionally, the addition of YCM was effective in prolonging shelf-life by increasing the oxidative stability of YCM-based cookies, indicated by the lower peroxide value (0.1 mEq O2·kg–1) after 4 months of storage compared to the regular cookies. YCM powder can be an alternative source of antioxidants, and it may be possible to include YCM powder in cookies and other pastries to increase their nutritional value and antioxidant activity to produce functional food

Enhancement of phenolic profile and antioxidant activity of black glutinous rice (Oryza sativa var. glutinosa) due to tape fermentation

Journal Article
Azkia, M.N, Cahyanto, M.N, Mayangsari Y, Briliantama A, Palma M, Setyaningsih W

Abstract: Black glutinous rice (BGR) naturally contains phenolic compounds in free and conjugated form or bound to the structural matrix, which may affect its bioactivity. The degradation of the matrix cell wall by microbial activity during fermentation may endorse the release of bound phenolic compounds and enhance the availability of the compounds. This study aimed to assess the impact of tape fermentation on the profile of individual phenolic compounds and the antioxidant activity of BGR. Fermentation up to 72 h at 30 °C was performed by adding the cooked BGR with a tape starter (0.3% w/w). The fermentation for over 48 h increased free phenolic compounds while altering the composition of free, free-conjugated, and bound phenolic compounds. In comparison to the initial material, significant enhancement in the level of protocatechuic aldehyde (13 × ), caffeic acid (2 × ), vanillic acid (1 × ), vanillin (1 × ), p-coumaric acid (1 × ), ferulic acid (1 × ), and isoferulic acid (1 × ) was acknowledged in the final fermented product while sinapic and 2,4,6-trihydroxybenzoic acid was merely detected after 24 h of fermentation. As a result, antioxidant activity was also increased notably (p < 0.05) starting from 48 h of fermentation, while principal component analysis revealed that the activity was strongly contributed by 2,4,6-trihydroxybenzoic acid. Henceforth, the resulting tape fermentation of BGR can be considered a source of phenolic compounds that provide high antioxidant activity.

Optimization of a New Ultrasound-Assisted Extraction Method of Caffeic Acid from the Aerial Parts of Coriandrum sativum by Using Experimental Design and Ultra-Performance Liquid Chromatography

Journal Article
Messaoudene, L, Palma, M, Carrera, C, Setyaningsih, W; Hazzid, M, Djebbar, R

Abstract: Coriander (Coriandrum sativum L.) is among the most widely used medicinal and aromatic plants. It is well known for its multiple health benefits, most of which are correlated with its phenolic composition. Four phenolic compounds were identified in the extracts of aerial parts of coriander extracts, including caffeic acid, isoquercitrin, quercetin-3-O-glucuronide, and rutin. Caffeic acid was the major compound in the extracts. A Box–Behnken Design (BBD) was employed in conjunction with the response surface methodology (RSM) to develop an ultrasound-assisted extraction method for the determination of phenolic compounds in the aerial parts of coriander using the level of caffeic acid as the target response. The following working variables were evaluated: methanol level in the extraction solvent, temperature, sonication time, and liquid-to-solvent ratio. It was found that the methanol concentration is the most significant factor that influences the recovery of caffeic acid. The optimal extraction conditions were: 10 min as the extraction time, 70 °C as the temperature, 50% for methanol in water as the solvent, and 6.51 mL of solvent per gram of sample. The repeatability and reproducibility were calculated and RSD values below 6% were obtained in both cases. The new method was employed for the extraction of real coriander samples and it is suggested that this method could potentially be applied for quality control analyses.

Optimization of microwave-assisted fish oil extraction from Patin (Pangasius micronemus) using Response Surface Methodology-Box Behnken Design (RSM-BBD)

Journal Article
Rosiana A, Setyaningsih W, Carrera C, Palma M, Rohman A, Riyanto S

Abstract: Patin (Pangasius micronemus) fish oil is rich in omega-3 fatty acids such as α-linolenic acid and eicosapentaenoic. To obtain the benefits of omega-3, it is necessary to develop an efficient extraction method for patin oil (PO). This study aimed to develop the microwave-assisted extraction (MAE) method for fish oil from patin. Three independent variables with three levels were priorly evaluated using Box–Behnken design, including temperature (30, 60, 90°C), solvent composition (30, 60, 90% of ethyl acetate in methanol), and solvent to sample ratio (10:1, 15:1, 20:1). The significant variables were then optimized by response surface methodology (RSM). A second order quadratic model of RSM suggested an extraction at 60°C with 56% ethyl acetate in methanol as an extraction solvent and a solvent-to-sample ratio of 20:1. A kinetics study under the optimum MAE conditions approved the complete recovery (38.84%) starting at 15 min of extraction time. The high precision of the MAE process was confirmed by the coefficient of variation less than 3%. Additionally, the microwave-produced fish oil was characterized by gas chromatography-mass spectrometry (GC-MS) to contain α-linolenic acid and eicosapentaenoic as omega-3. Henceforth, it has been demonstrated that microwave-assisted fish oil extraction developed in this study is efficient for high-quality PO production.

Optimized Ultrasound-Assisted Extraction of Psidium laruotteanum Roots: A Concentrated Source of Piceid from the Brazilian Savanna

Journal Article
Takao, Leandro Kenji; ; Setyaningsih, W; Gualtieri, Sonia C.J; Rodriguez, Ana Ruiz; Varela, Rosa M; Palma, Miguel

Abstract: Psidium laruotteanum is a subshrub Myrtaceae, native to the Brazilian savanna, and distributed in Central and South America. Here, the health beneficial compound piceid (reservatrol-3-mono-D-glucoside) is described for the first time in the genus Psidium. The ultrasound-assisted extraction from P. laruotteanum roots was optimized by a response surface methodology. A central composite design was used to assess six extraction variables. Optimal extractions were obtained with 62.5% MeOH in water at 69.1 C, amplitude of 70%, cycle of 0.5 s􀀀1, pH 5.7 and solvent–solid ratio of 40:1, for 10 min. The method showed excellent recovery (90%–95%) and precision (CV: 0.69% for repeatability and 0.43% for intermediate precision). This species appears as the most concentrated plant source of piceid reported to date (29.15 mgg􀀀1 in dry roots). Additionally, a very fast extraction (5 min) can afford 96% recovery of piceid from Psidium laruotteanum roots. It could be used as a natural and cheap source of piceid for several applications.

Ultrasound-Assisted Anthocyanins Extraction from Pigmented Corn: Optimization Using Response Surface Methodology

Journal Article
Nurkhasanah, Annisa; Fradad, Titouan; Carrera, Ceferino; Setyaningsih, W; Palma, Miguel

Abstract: This study aimed to determine the optimal UAE conditions for extracting anthocyanins from pigmented corn using the Box–Behnken design (BBD). Six anthocyanins were identified in the samples and were used as response variables to evaluate the effects of the following working variables: extraction solvent pH (2–7), temperature (10–70 ◦C), solvent composition (0–50% methanol in water), and ultrasound power (20–80%). The extraction time (5–25 min) was evaluated for complete recovery. Response surface methodology suggested optimal conditions, specifically 36% methanol in water with pH 7 at 70 ◦C using 73% ultrasound power for 10 min. The method was validated with a high level of accuracy (>90% of recovery) and high precision (CV < 5% for both repeatability and intermediate precision). Finally, the proposed analytical extraction method was successfully applied to determine anthocyanins that covered a wide concentration range (36.47–551.92 mg kg−1) in several pigmented corn samples revealing potential varieties providing more health benefits.

Enhanced Differentiation of Wild and Feeding Civet Coffee Using Near-Infrared Spectroscopy with Various Sample Pretreatments and Chemometric Approaches

Journal Article
Prajna, Deyla; Alvarez, Maria; Sepulveda, Marta Barea; Calle, Jose; Suhandy, Diding; Setyaningsih, Widiastuti; Palma, Miguel

Civet coffee is the world’s most expensive and rarest coffee bean. Indonesia was the first country to be identified as the origin of civet coffee. First, it is produced spontaneously by collecting civet feces from coffee plantations near the forest. Due to limited stock, farmers began cultivating civets to obtain safe supplies of civet coffee. Based on this, civet coffee can be divided into two types: wild and fed. A combination of spectroscopy and chemometrics can be used to evaluate authenticity with high speed and precision. In this study, seven samples from different regions were analyzed using NIR Spectroscopy with various preparations: unroasted, roasted, unground, and ground. The spectroscopic data were combined with unsupervised exploratory methods (hierarchical cluster analysis (HCA) and principal component analysis (PCA)) and supervised classification methods (support vector machine (SVM) and random forest (RF)). The HCA results showed a trend between roasted and unroasted beans; meanwhile, the PCA showed a trend based on coffee bean regions. Combining the SVM with leave-one-out-cross-validation (LOOCV) successfully differentiated 57.14% in all sample groups (unground, ground, unroasted, unroasted–unground, and roasted–unground), 78.57% in roasted, 92.86% in roasted–ground, and 100% in unroasted–ground. However, using the Boruta filter, the accuracy increased to 89.29% for all samples, to 85.71% for unground and unroasted– unground, and 100% for roasted, unroasted–ground, and roasted–ground. Ultimately, RF successfully differentiated 100% of all grouped samples. In general, roasting and grinding the samples before analysis improved the accuracy of differentiating between wild and feeding civet coffee using NIR Spectroscopy

Optimization of Ultrasound-Assisted Cold-Brew Method for Developing Roselle (Hibiscus sabdariffa L.)-Based Tisane with High Antioxidant Activity

Journal Article
Larasati, Intan Dewi; Oktaviani, Nurul Mutmainah Diah; Lioe, Hanifah Nuryani; Estiasih, Teti; Palma, Miguel; Setyaningsih, Widiastuti

Edible flowers, including roselle, contain a varied composition of phenolic compounds that may inhibit the oxidative stress mechanism. Roselle-based tisane with appealing sensory properties is commonly consumed worldwide. However, the conventional hot-brew method may ruin the stability of thermolabile phenolic compounds during the tisane preparation. Hence, this study aimed to develop a new alternative brewing method linked with the new cold-brew method, which involves a lower temperature and applying ultrasound to maximize the extraction of phenolic compounds and to avoid degradation during the tisane preparation. The brewing factors, including particle size (10, 20, 30 mesh), temperature (4, 15, 26 °C), time (10, 20, 30 min), and ultrasound amplitude (20, 60, 100% of the maximum amplitude) have been optimized simultaneously using Box–Behnken design in conjunction with response surface methodology. Seven major phenolic compounds were identified by HPLC-DAD and classified into hydroxycinnamic acid derivatives (HCA) and flavonoids. The optimum extraction condition to reach the highest level of the studied phenolic compounds was set to brew roselle with particle size of 30 ± 3.25 mesh at 26 ± 1.32 °C for 18 ± 2.00 min applying 78 ± 5.64% ultrasound amplitude. This method successfully extracted almost all HCA and flavonoid during the first cycle with less than 10% CV and provided higher antioxidant activity in terms of DPPH (IC50 9.77 ± 0.01 µg mL−1), ABTS (IC50 8.05 ± 0.02 µg mL−1), and FRAP (IC50 10.34 ± 0.03 µg mL−1) than the roselle tisane prepared using the conventional method. Additionally, the resulting cold-brew product was stable for up to five days of storage.

Response Surface Methodology Optimization for Analytical Microwave-Assisted Extraction of Resveratrol from Functional Marmalade and Cookies

Journal Article
Widiastuti Setyaningsih, María del Cisne Guamán-Balcázar, Nurul Mutmainah Diah Oktaviani, Miguel Palma
Foods, Volume 12, Issue 2, December 2022

A novel analytical method based on microwave-assisted extraction has been successfully optimized and validated to determine resveratrol from functional marmalade and cookies. The optimization was performed using a Box–Behnken design with three factors: solvent composition (60−100% and 10−70% methanol in water for marmalade and cookies, respectively), microwave power (250−750 W), and solvent-to-solid ratio (20:5−60:5). The main and quadratic effects of solvent composition significantly contributed to the recovery values (p < 0.005) for both kinds of samples. Additionally, the solvent-to-solid ratio and the quadratic effect of microwave power also influenced the resveratrol recovery from functional marmalade. Hence, the optimum condition for resveratrol extraction from marmalade (80% methanol, 500 W, solvent-to-solid ratio 40:5) and cookies (80% methanol, 250 W, solvent-to-solid ratio 20:5) was proposed. The extraction kinetics (5−30 min) was then studied to clarify the complete recovery of resveratrol from the food matrices resulting in 15 min as the optimum extraction time. The developed method showed a satisfactory precision indicated by the coefficient of variation (CV) lower than 5.70% for both repeatability and intermediate precision. To check the matrix effects, the developed MAE procedures were applied to a number of commercial marmalade and cookies. The high-fat and fiber cookies resulted in overestimated values due to the interfering ingredients. As a final point, the methods successfully measured the stability of naturally present resveratrol in grape-derived products while preparing functional marmalade and cookies.

Optimization of Ultrasound-Assisted Extraction (UAE) for Simultaneous Determination of Individual Phenolic Compounds in 15 Dried Edible Flowers

Journal Article
Asadin Briliantama, Nurul Mutmainah Diah Oktaviani, Sitti Rahmawati, Widiastuti Setyaningsih, Miguel Palma
Horticulutrae, Volume 8, Issue 12, 19 December 2022

Nowadays, dried edible flowers have become one of the eating habits of a healthy lifestyle. The most common way to consume dried flowers is via infused water (tisane). A number of studies on dried edible flowers have reported antioxidant activities mainly due to their phenolic compounds. This work has developed a new extraction method using ultrasound technology to determine phenolic compounds in 15 widely consumed edible flowers. Several extraction factors including pulse duty cycle (0.2, 0.6, 1.0 s−1), temperature (10, 40, 70 °C), solvent-to-sample ratio (10:1, 20:1, 30:1 mL of solvent g −1 of sample), and solvent composition (0, 25, 50% methanol in water) have been optimized based on a Box–Behnken design coupled with response surface methodology. UPLC-PDA has been employed to quantify 12 major phenolic compounds (2,4,6-trihydroxy benzoic acid, protocatechuic acid, protocatechuic aldehyde, p-hydroxybenzoic acid, caffeic acid, vanillic acid, epicatechin, p-coumaric acid, ferulic acid, quercetin-3-rutinose, iso-ferulic acid, and quercetin-3-glucoside) in the extracts. The optimum extraction conditions for a 1 g sample were 30 mL of solvent (28% methanol in water) at 42 °C with 1.0 s−1 of pulse duty cycle. Based on the kinetic study, the optimal extraction time was 10 min. The method was validated with high precision (CVs of repeatability and intermediate precision were lower than 7%) and high accuracy (recovery higher than 90%). Additionally, the proposed ultrasound-assisted extraction was successfully applied in the determination of phenolic compounds in 15 dried edible flowers.

A Rapid Method for Authentication of Macroalgae Based on Vis-NIR Spectroscopy Data Combined with Chemometrics Approach

Journal Article
Frysye Gumansalangi, Jose L. P. Calle, Marta Barea-Sepúlveda, Manikharda, Miguel Palma, Lideman, Mohamad Rafi, Andriati Ningrum, Widiastuti Setyaningsih
Water, Volume 15, Issue 1, 28 December 2022

Macroalgae are an essential basic ingredient for many industries from which numerous derived products with great economic value are manufactured. Each macroalga has a unique composition that might provide specific physical and chemical information that can be used as markers for authentication. Their compositions may differ depending on different factors, including geographical regions. Unsupervised exploratory techniques, namely principal component analysis (PCA) and hierarchical cluster analysis (HCA), and nonparametric supervised methods including support vector machines (SVMs) and random forests (RFs), were applied to the Vis-NIR spectroscopic data to standardize the quality of macroalgae based on three regional zones in Indonesia (Western, Central, Eastern). A total of 35 macroalgae samples from six islands in Indonesia were analyzed. The PCA and HCA results present a tendency for the samples to be distributed and clustered according to the type of their species. Meanwhile, the SVM successfully classified samples based on their regional zones, and when combined with five-fold cross-validation, acquired an accuracy of 82%. The RF model algorithm obtained an accuracy of 100%, 80%, and 82% for the training, test, and five-fold cross-validation, respectively.

Validation of a Rapid Ultrasound-Assisted Extraction Coupled with Anion Exchange Chromatography Method for the Determination of D-Psicose in Raisin Matrices

Journal Article
Fahmi Maulana Zulkarnaen ,Widiastuti Setyaningsih, Miguel Palma
Separations 2022, Volume 9, Issue 12, 2 December 2022

D-psicose is a rare sugar, used as an alternative to the natural sweetener that provides 70% of sucrose’s sweetness, with low-calorie absorption in the human body. Considering the health-beneficial effect and high availability of D-psicose in raisins (the world’s most consumed dried fruit), it is of interest to establish a reliable analytical method to determine D-psicose content in these matrices. Herein, a new method for determining D-psicose content in raisins, using ultrasound-assisted extraction in conjunction with anion exchange chromatography with pulsed amperometric detection (UAE-HPAEC-PAD) systems, has been developed and validated. The stability of D-psicose and its precursor was priorly assessed by applying a specific ultrasound power (100 W) and pulse duty cycle (0.5 s−1), with varying extraction temperatures (10, 25, 40, 55, 70, and 85 °C) and times (5, 10, 15, 20, and 25 min). The method was validated with high linearity (R2 > 0.999), accuracy (89.78–101.06%), and precision (4.8% intra-day and 4.34% inter-day). A number of raisin products were checked during the method applicability assessment. A concentration of 520 mg kg−1 was found in a specimen of commercial raisin matrix.

Optimization of Ultrasound-Assisted Extraction from Young Coconut Mesocarp in the Rapid Extraction of Phenolic Compounds and Antioxidant Activity

Journal Article
Lulum Leliana, Widiastuti Setyaningsih, Miguel Palma, Supriyadi, Umar Santoso
Agronomy, Volume 12, Issue 11, Pages 1-12

Abstract: Coconut (Cocos nucifera) fruit has attracted consumer interest due to its health benefits,
especially the consumption of coconut water produced from 6–7-month-old coconut fruit. Consequently,
by-products from young coconut fruit are also being expanded, including coconut mesocarp
containing phenolic compounds. Therefore, this study aimed to provide new applications for young
coconut mesocarp wastes. Specifically, optimizing the ultrasound-assisted extraction (UAE) of phenolic
compounds from coconut mesocarp using a Box–Behnken design in conjunction with response
surface methodology (RSM). The effects of three extraction variables, such as temperature (10–70 C),
solvent-to-sample ratio (20:1–10:1 mL/g), and pulse duty cycle (0.4–1.0/s) were examined on
the level of total phenolic content (TPC) and antioxidant activity of the extract. Subsequently, the
optimum UAE condition was predicted using RSM models with coefficients of determination (R2)
higher than 0.94, low prediction errors (less than 2.34), and non-significant lack-of-fit values (p < 0.05)
for the two responses. Extraction time was evaluated through kinetic (5 to 25 min) studies applying
the optimum extraction temperature (70 C), solvent-to-sample ratio (20:1 mL/g), and pulse duty
cycle (0.55/s). An efficient extraction was achieved within 5 min, resulting in an extract with
47.78 1.24 mg GAE 100/g DW for the total phenolic compounds and high antioxidant activity
(87.28 1.01% DPPH). Extraction by ultrasound was then concluded to facilitate a fast extraction
rate with high reproducibility (coefficients of variation were less than 3% in the levels of antioxidant
activity and phenolic compounds).

A new FTIR method combined with multivariate data analysis for determining aflatoxins in peanuts (Arachis hypogaea)

Journal Article
Aan Sulistiawan, Widiastuti Setyaningsih , Abdul Rohman.
Journal of Applied Pharmaceutical Science Vol. 12(07), pp 199-206, July, 2022

A rapid method based on Fourier transform infrared (FTIR) spectroscopy with multivariate data analysis was developed to identify and quantify aflatoxin contamination in peanuts. This technique was proposed over the chromatographic method due to faster, practical, and less reagent consumption as it is considered a non-destructive method. The spectra of samples were scanned using an FTIR spectrophotometer with attenuated total reflectance in the mid-infrared region (4,000−475 cm−1). Chemometric techniques for classification purposes using principal component analysis (PCA) and multivariate calibrations of partial least square (PLS) regression were used to establish the quantitative model and predict the levels of aflatoxin contamination in peanuts. PCA was successfully applied to classify the whole peanuts, those without shells, and those without shells and testa, based on PC1 and PC2 score plots. The determination of aflatoxin contamination levels using high-performance liquid chromatography coupled with a fluorescence detector (HPLC-FD) showed that 4 out of 20 samples (20%) resulted in 1.272–9.585 µg kg−1 for AFB1 and 0.448–2.943 µg kg−1 for AFB2. The method performance of aflatoxin analysis by HPLC-FD was validated by high coefficients of determination (R2 > 0.999) and low coefficients of variation (CV = 1.68−2.94%). Furthermore, PLS regression based on FTIR spectra at a selected wavenumber region (1,800−800 cm−1) provided a satisfactory prediction of aflatoxin contamination in peanut samples, including aflatoxin B1 [R2= 0.9995; root mean square error of calibration (RMSEC)= 0.1180; root mean square error of prediction (RMSEP) = 0.3000)] and aflatoxin B2 (R2 = 0.9999; RMSEC = 0.0119; RMSEP = 0.0595). Therefore, the developed method can be an analytical tool to identify the occurrence of aflatoxins in peanuts in the market.

Experimental Design-Assisted Optimization of Chromatograhpic Method for the Simultaneous Quantitation of Phenolic Compounds in Dried Flowers Extract

Journal Article
Mira Thorvaldsson, Nurul Mutmainah, Asadin Briliantama, Sitti Rahmawati, Ashari Priyanto, Pargiyanti, Widiastuti Setyaningsih
Journal of Functional Food and Nutraceutical, Volume 3, No. 2, February 2022, Pages 83-91

Dual Response Optimization of Ultrasound-Assisted Oil Extraction from Red Fruit (Pandanus conoideus) Recovery and Total Phenolic Compounds

Journal Article
Endah Prasetia Susanti, Abdul Rohman, Widiastuti Setyaningsih
Agronomy, Volume 12, Issue 523, February 2022, Pages 1-16

Abstract: Red fruit oil (RFO) is a high-value oil that contains functional compounds, mainly phenolic compounds, providing antioxidant activity. Therefore, an optimal extraction method is essential to recover the RFO and phenolic compounds simultaneously. This research aimed to optimize the ultrasound-assisted extraction (UAE) for oil from red fruit using the Box-Behnken design combined with response surface methodology. The studied UAE factors, including sample-to-solvent ratio (1:3,1:2, and 1:1 g mL^-1), extraction temperature (60, 75, and 90 C), and pulse duty-cycle (0.20, 0.50, and 0.80 s????1). Analysis of variance revealed that the three studied factors significantly influenced the recovered RFO, while the level of total phenolic compounds in the extracts was defined merely by extraction temperature (p < 0.05). These significant factors were then included in the optimization models (R2 > 0.99, lack-of-fit p > 0.05). The proposed UAE setting by the multiresponse optimization was an extraction temperature of 67 C, a pulse duty-cycle of 0.50 s^-1, and a sample-to-solvent ratio of 1:2.5 g mL^-1. Subsequently, the extraction kinetic was evaluated, confirming full recovery at 60 min of extraction time. The developed method was then applied to extract six red fruit clones. Mbarugum clones provided high RFO recovery (9.60%), with an uppermost total phenolic compound of (42.63 mg GAE g^-1) among the six red fruit clones. Additionally, the resulting RFO showed eminent antioxidant activities, indicated by excellent values of IC50 DPPH (37.69 mg L^-1), IC50 FIC (30.43 mg L^-1), FRAP reducing power (63.55 mg AAEA g^-1), and IC50 ABTS (93.88 mg L^-1). In contrast with a wet rendering method, UAE enhanced the RFO recovery by 53.02%, resulting in a higher level of total phenolic compounds. Henceforth, the proposed UAE method is a promising technique to substitute conventional oil production in the food and pharmaceutical industries.

The Use of Chemometrics for Classification of Sidaguri (Sida rhombifolia) Based on FTIR Spectra and Antiradical Activities

Journal Article
Abdul Rohman, Asefin Nurul Ikhtiarini, Widiastuti Setyaningsih, Mohamad Rafi, Nanik Siti Aminah, Muhamad Insanu, Irnawati Irnawati, Djoko Santosa
Indonesian Journal of Chemistry, Volume 21, Issue 6, Pages 1568-1576

Abstract: Sidaguri (Sida rhombifolia) is one of the herbal components used in traditional medicine. The application of chemometrics in the standardization of herbal medicine is common. The objective of this study was to classify Sidaguri from different regions based on FTIR spectra with chemometrics of principal component analysis (PCA) and to correlate the antioxidant activities with FTIR spectra using the multivariate calibration of partial least square regression (PLSR). The extraction of Sidaguri powder was performed using ultrasound-assisted extraction (UAE) at optimum conditions. The obtained extracts were subjected to antiradical scavenging activities using DPPH (2,2’-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) radicals. The PCA result shows that Sidaguri from different regions could be separated using 14 wavenumbers of FTIR spectra based on the PCA’s loading plot. PLSR regression using the second derivative FTIR spectra at wavenumbers of 3662–659 cm–1 could predict radical scavenging activities (RSA) of Sidaguri with R2 values of 0.9636 and 0.9024 for calibration and validation models, with RMSEC and RMSEP values of 1.45% and 2.65%, respectively. It can be concluded that FTIR spectra treated by PCA were reliable for classifying Sidaguri from different regions. At the same time, PLSR was accurate and precise enough to predict the RSA of Sidaguri.

 

A microwave-based extraction method for the determination of sugar and polyols: Application to the characterization of regular and peaberry coffees

Journal Article
Widiastuti Setyaningsih, Andika Wicaksono Putro, Rohmah Nur Fathimah, Kiki Adi Kurnia, Noviyan Darmawan, Brian Yulianto, Prastika Krisma Jiwanti, Ceferino A. Carrera, Miguel Palma
Arabian Journal of Chemistry, Volume 15, March 2022, Pages 1-10

Abstract The brewing properties of coffee products are defined by the chemical composition in the bean, including sugars and polyols. Some factors, such as coffee species and roasting, may affect the level of these compounds in the bean. A new analytical microwave-assisted extraction (MAE) method has been developed to extract sugars and polyols from the coffee bean. The studied extraction conditions for the MAE were temperature (30–80 C), solvent composition (0–50% ethanol in water), and solvent-to-sample ratio (10:1–30:1 mL solvent per g sample). A Box-Behnken design was applied to study the effect of extraction variables, and subsequently, the influential variables were optimized by response surface methodology (RSM). In addition to the main effect of the solvent-to-sample ratio, all quadratic effects significantly influenced (p < 0.05) the recovery of sugars and polyols from the coffee beans. RSM suggested the optimized MAE conditions: temperature 52 C, ethanol concentration in water 18.5%, and solvent-to-sample ratio 17:1. Under the optimum condition, a kinetics study confirmed that 15 min showed high precision and accuracy of the developed method. Ultimately, a real sample application of the developed MAE revealed that the new method successfully described the composition of sugars and polyols in regular and peaberry coffee beans. Additionally, the method also effectively characterized the green and roasted Arabica and Robusta coffee beans.

 

Oligo-Glucomannan Production From Porang (Amorphophallus oncophyllus) Glucomannan By Enzymatic Hydrolisis Using β-Mannanase

Journal Article
Anggela, Widiastuti Setyaningsih, Santad Wichienchot, Eni Harmayani
IFNP, Vol. 17, No. 1 (2020)

Abstract: Porang (Amorphophallus oncophyllus) is an indigenous tuber of Indonesia that rich in glucomannan. An alternative approach to produce porang oligo-glucomannan (POG) as prebiotic from porang glucomannan (PGM) was made by enzymatic hydrolysis using β-mannanase. This study aimed to produce POG under optimal conditions by controlled enzymatic hydrolysis process. The PGM flour contained 96.12% of indigestible carbohydrates. The optimum condition of enzymatic hydrolysis producing the highest reducing sugar was as follows: temperature 37 °C, pH 5.5, a ratio of enzyme to the substrate (E/S) 1:1000, and reaction time 4 h. HPLC analysis confirmed that 99.45% of the resulting POG consisted of oligosaccharides with a degree of polymerization (DP) 3. Hence, the PGM utilized in this study has been proven as a potential substrate for POG production. Additionally, the resulting POG was considered as a functional ingredient due to has prebiotic potential.

Optimasi Sintesis Hydroxypropyl Methyl Cellulose (HPMC) dari Kulit Koro Pedang Putih (Canavalia ensiformis L. (DC)

Journal Article
Yanti Nopiani, Agnes Murdiati, Widiastuti Setyaningsih
agriTECH, 41 (4) 2021, 395-407

Abstract: Kulit koro pedang putih dapat digunakan sebagai sumber selulosa. Salah satu alternatif untuk meningkatkan aplikasi selulosa adalah dengan memodifikasi selulosa menjadi produk turunan selulosa yaitu Hydroxypropyl Methyl Cellulose (HPMC). Tujuan dari penelitian ini adalah melakukan optimasi terhadap sintesis dan karkaterisasi HPMC dari selulosa kulit koro pedang putih. Proses optimasi didahului dengan kajian literatur untuk menentukan kisaran titik percobaan dengan variabel terikat berupa molar subtitusi (MS) dan Derajat Subtitusi (DS). Diperoleh titik percobaan dengan variasi konsentrasi NaOH (5, 22,5, dan 40%), variasi Dimetil Sulfat (DMS) (40, 80, dan 120%), dan variasi Proilen Oksida (PO) (80, 120, dan 160%). Kemudian optimasi sintesis HPMC dilakukan dengan mengunakan Box-Behnken design (BBD) lalu dianalisis menggunakan Response Surface Methodology (RSM) Berikutnya HPMC dikarakterisasi meliputi molar subtitusi (MS), Derajat Subtitusi (DS), water holding capacity (WHC), oil holding capacity (OHC), lightness, rendemen, kristalinitas dan spektra FT-IR untuk mengetahui gugus fungsi HPMC. Hasil optimasi sintesis HPMC dari selulosa kulit koro pedang putih berdasakan RSM diperoleh titik optimum pada konsentrasi NaOH 23,11%, DMS 43,4% dan PO 81,8%. dengan karakterisasi kadar air 9,04% (wb); MS 0,15; DS 1,18; WHC 2,20 g/g; OHC 2,09 g/g; lightness 90,93; rendemen 114,78% dan kristalinitas 64%. Spektra FT-IR HPMC koro pedang putih terbaca pada bilangan gelombang 2924 cm-1 (CH dan CH2Streching), 1373 cm-1 (CH3Bonding), 1118 cm-1 (C-O-C), 1319 cm-1 (O-H Plane Bonding) dan 848, 68 cm-1 (C-O-C pada 1,4 β glikosidic linkage) yang merupakan ciri khas dari gugus fungsional HPMC.

An ultrasound-based technique for the analytical extraction of phenolic compounds in red algae

Journal Article
Venansius G.P., Selma Mutiarahma, Weni Chaniago, Puji Rahmadi, Dedy Kurnianto, Chusnul Hidayat, Caferino Carrera, Miguel Palma, Widiastuti Setyaningsih
Arabian Journal of Chemistry (2022) 15, 103597

Abstract: Phenolic compounds are bioactive compounds that are also naturally found in red algae. To determine the level of these compounds in the red algae, spectroscopic or chromatographic determination was applied over the liquid extracts. Therefore, a prior extraction method is needed. The presented study aimed to develop the analytical ultrasound-assisted extraction (UAE) method to extract phenolic compounds from red algae. A Box–Behnken design (BBD) based on five factors included solvent composition (50–90% ethanol in water), extraction temperature (10–60 C), ultrasonic power (20–100%), pulse duty-cycle (0.2–1.0 s1), and solvent-to-sample ratio (10:1 to 30:1) was used to evaluate the effects of the studied factors. Subsequently, response surface methodology (RSM) was performed to define the optimum extraction condition to recover phenolic compounds from the alga matrices. The UAE condition suggested by RSM was: ultrasonic power 100%, pulse duty-cycle 1 s1, temperature 52.5 C, extraction solvent 50% ethanol in water, and solvent-to sample ratio 30:1. Kinetic studies confirmed 10 min to provide comparable recovery (p>0.05) than any longer extraction time. The acceptable values validated the developed method for repeatability (CV, 4.8%) and intermediate precision (CV, 5.7%). In addition, the accuracy of the method suggested a complete recovery for two extraction cycles. Furthermore, the method has successfully.

An Information and Marketing System for Agrocomplex Sector Based on Reliable Digital Technology as a Supporting Capacity for Food Security in a Force Majeure Event

Journal Article
Endang Sulistyaningsih, Valentina Dwi Suci Handayani, Widiastuti Setyaningsih, Wahdan Fitriya, Arif Wahyu Widada, Candra Aryudiawan, Taufan Alam, Imas Masithoh Devangsari
Indonesian Journal of Community Engagement, Volume 7, No. 3, September 2021, Pages 190-194

The pandemic situation experienced by the world is currently threatening food security in various regions including Indonesia. As a strategic step to reduce the impact on the agricultural sector, it is necessary to accelerate information to support the stability of production and also to distribute production results. Digital media in agriculture are needed to provide education with a variety of contents, such as technical production, marketing, and agricultural finance information. The Faculty of Agriculture of Universitas Gadjah Mada (UGM) has supported digital information dissemination through the Desa Apps application. Desa Apps aims to be an application platform that provides agricultural extension and develops digital farmer communities. The information that has been carried out by Desa Apps so far includes agrocomplex sector (agriculture, animal husbandry, and fisheries) problems related to cultivation methods and obstacles faced in the land such as attack by plant pest and disease. As a digital media, the Desa Apps has educated farmers and aquaculturists. This study evaluates whether farmers and aquaculturists under the auspices of the Faculty of Agriculture are interested in being able to get an education and at the same time be able to market their product directly to consumers with one application, the Desa Apps. The findings of the study showed that the pandemic period greatly affected the incomes of the assisted target community, and they required media that can help market their product. The decline in turnover due to pandemic conditions reached 70% with an average decrease of 39.2%. The method of selling products has changed from conventional to online sales by 20% of the total respondents. Changes in sales methods occurred due to distribution limitations due to regional and activity restrictions during the pandemic. Therefore, it is necessary to optimize the Desa Apps application as a digital media to support food security.

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

Journal Article
Selma Mutiarahma, Venansius G. P. Putra, Weni Chaniago, Ceferino Carrer, Sri Anggrahini, Miguel Palma, and Widiastuti Setyaningsih
Mutiarahma, Selma; Putra, Venansius Galih Perkasa; Chaniago, Weni; Carrera, Caferino; Palma, Miguel; Setyaningsih, W. 2021. UV-Vis Spectrophotometry and UPLC-PDA Combined with Multivariate Calibration for Kappapycus alvarezii (Doty) Doty ex Silva Standardization Based on Phenolic Compounds. Scientia Pharmaceutica, 89(47): 1-13

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 com pounds. 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 (R2) > 0.96 in the calibra‐ tion 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 com‐ ponent 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.

Microwave-Assisted Extraction of Phenolics from Hibiscus sabdariffa Flowers: Method Development and Validation

Conference Paper
Rohmah Nur Fathimah, Widiastuti Setyaningsih, Ceferino Carrera, and Miguel Palma
Fathimah, R.N., Carrera, C., Setyaningsih, W., Palma, M. Microwave-assisted Extraction of Hibiscus sabdariffa Antioxidants: Method Development and Validation. 1st International Electronic Conference on Food Science and Functional Foods, November 10-25, 2020, https://sciforum.net/conference/foods_2020.

Phenolics are bioactive compounds already reported in Hibiscus sabdariffa (roselle). The present study reports the development and validation of the analytical microwave-assisted extraction (MAE) method for the determination of phenolic compounds in roselle flowers to establish their positive contribution to human health. Prior to the optimization, a study for identifying phenolic compounds revealed that chlorogenic acid, protocatechuic acid, caffeic acid, and rutin were presented in Roselle. Three factors affecting MAE, viz. temperature, solvent composition, and sample to solvent ratio, were optimized employing a Box-Behnken Design (BBD) in conjunction with response surface methodology (RSM). The maximum extraction recovery was achieved using the extraction temperature of 68 °C, solvent composition of 59% MeOH in water, and 20:1 solvent to sample ratio. The kinetics experiment confirmed full recoveries (92–119%) at 15 min. Subsequently, method validation showed a satisfactory result, including low detection limits from 0.219 mg L−1 (caffeic acid) to 0.374 mg L−1 (chlorogenic acid). Both precisions and accuracy met the acceptances by AOAC. Finally, the method was successfully applied to quantify phenolics in the two most common varieties of Roselle. Chlorogenic acid was found as the main phenolic compound in both varieties

Optimization of ultrasound-assisted extraction and the antioxidant activities of Sidaguri (Sida rhombifolia)

Journal Article
Ikhtiarini, Asefin N., Setyaningsih, W., Rafi, M., Aminah, Nanik S., Insanu, M., Irnawati, I., Rohman, Abdul
Journal of Applied Pharmaceutical Science, 11(08), 070-076.

Sidaguri (Sida rhombifolia) is widely distributed in tropical and subtropical regions and has been reported to
possess many bioactive compounds that are beneficial for human health, including polyphenolics having antioxidant
activities. The objective of this study was (1) to optimize ultrasound-assisted extraction (UAE) of Sidaguri and (2)
to evaluate antioxidant activities of extracts resulting from the optimized condition using radical scavenging assay of
2,2′-diphenil-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid. The optimum UAE
conditions obtained were solvent to solid ratio of 26:1, temperature of extraction of 45.45°C, methanol concentration
of 42%, power of sonication of 86%, and time of extraction of 5 minutes. Sidaguri obtained from locations of Kretek
and Ngemplak had the highest radical scavenging activity accounting for 83.69% ± 0.30% and 82.95% ± 0.49%,
respectively.

Effect of different cooking methods on chemical composition, nutritional values, and sensory properties of Jack bean (Canavalia ensiformis) tempe

Journal Article
Purwandari, F.A., Annisa, E.D.N., Rachmawati, A.T., Puspitasari, D., *Wikandari, R., Setyaningsih, W., Ningrum, A. and Sardjono
Food Research 5 (3) : 327 - 333

Having high protein content, jack bean is a promising resource for making tempe, a traditional fermented food commonly consumed in Indonesia. The current research aimed to investigate the effect of different cooking methods (boiling, steaming, frying, and baking) on a chemical, nutritional, and sensory aspects of jack bean tempe. The results showed that the highest protein content and antioxidant activity of the processed tempe were obtained by steaming method with a value of 380.3 g/kg and 57.8%, respectively. The steaming process could maintain protein and mineral contents as well as the antioxidant activity. The boiled tempe had six time lower of phytic acid compared to that of control. The baking and frying process increased five to six-fold of the fat content, while reduced the protein content by 30%. For the sensory evaluation, the baked tempe obtained the highest score. In conclusion, from a nutritional point of view, steaming is the best method for cooking jack bean tempe, while in term of sensory, baking is recommended.

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

Journal Article
Carrera, C.; Pastol, J.; Setyaningsih, W.; Ruiz-Rodríguez, A.; Ferreiro-González, M.; Fernández Barbero, G.; Palma, M.
Agronomy 2021, 11, 1053

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 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 s1). 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 C extraction temperature, 0.3 s1 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.

Analytical Methods for Determination of Non-Nutritive Sweeteners in Foodstuffs

Journal Article
Viki Oktavirina, Nadhila B. Prabawati, Rohmah Nur Fathimah, Miguel Palma, Kiki Adi Kurnia, Noviyan Darmawan, Brian Yulianto and Widiastuti Setyaningsih
Molecules 2021, 26, 3135.

Sweeteners have been used in food for centuries to increase both taste and appearance. However, the consumption of sweeteners, mainly sugars, has an adverse effect on human health when consumed in excessive doses for a certain period, including alteration in gut microbiota, obesity, and diabetes. Therefore, the application of non-nutritive sweeteners in foodstuffs has risen dramatically in the last decade to substitute sugars. These sweeteners are commonly recognized as high-intensity sweeteners because, in a lower amount, they could achieve the same sweetness of sugar. Regulatory authorities and supervisory agencies around the globe have established the maximum amount of these high-intensity sweeteners used in food products. While the regulation is getting tighter on the market to ensure food safety, reliable analytical methods are required to assist the surveillance in monitoring the use of high-intensity sweeteners. Hence, it is also necessary to comprehend the most appropriate method for rapid and effective analyses applied for quality control in food industries, surveillance and monitoring on the market, etc. Apart from various analytical methods discussed here, extraction techniques, as an essential step of sample preparation, are also highlighted. The proper procedure, efficiency, and the use of solvents are discussed in this review to assist in selecting a suitable extraction method for a food matrix. Single- and multianalyte analyses of sweeteners are also described, employing various regular techniques, such as HPLC, and advanced techniques. Furthermore, to support on-site surveillance of sweeteners’ usage in food products on the market, non-destructive analytical methods that provide practical, fast, and relatively low-cost analysis are widely implemented.

Edible Flowers: Antioxidant Compounds and Their Functional Properties

Journal Article
Nadhila B. Prabawati, Viki Oktavirina, Miguel Palma and Widiastuti Setyaningsih
Horticulturae. 2021; 7(4):66

Edible flowers have been widely consumed for ages until now. The attractive colors and shapes, exotic aroma, and delightful taste make edible flowers very easy to attain. Moreover, they also provide health benefits for consumers due to the unique composition and concentration of antioxidant compounds in the matrices. Knowing the bioactive compounds and their functional properties from edible flowers is necessary to diversify the usage and reach broader consumers. Therefore, this reported review could be useful for functional product development, engaging the discussed edible flowers. We present a comprehensive review of edible flower composition and the functional properties of their antioxidant compounds, mainly phenolics.

Phenolic compounds in olive oil by solid phase extraction – Ultra performance liquid chromatography – Photodiode array detection for varietal characterization

Journal Article
L. Deflaoui, W. Setyaningsih, M. Palma, A. Mekhoukhe, A. Tamendjari
Arabian Journal of Chemistry (2021) 14, 103102

An new method for the extraction of phenolic compounds from olive oil has been proposed before the chromatographic analysis by UPLC-PDA (Ultra-Performance Liquid Chromatography-PhotoDiode Array). Recoveries for 20 individual phenolic compounds were compared using three cartridges of solid-phase extraction including C-18, Diol, and DVB (DiVinylBenzene) based cartridges. Diol based cartridge produced the best retention efficiency. The optimized method uses a reduced amount of solvents and allows for the determination of most phenolics in oil samples in less than 10 min. A full discrimination among the most common olive oil varieties was achieved using the relative levels for specific phenolics in the sample. Therefore, based in this study, a fingerprint based on the relative levels for quercetin, luteolin, oleuropein and vanillic acid can be proposed to characterized varietal olive oils. In that way, a detailed composition about phenolic compounds of varietal olive oils would not be needed for discrimination purposes.

A microwave-based technique to determine saccharides and polyols contents in Spirulina (Arthrospira platensis)

Journal Article
Fathimah, R.N., Setyaningsih, W., Carrera, C., Astari, A.D., Mashitoh, R.E., Suryaningtyas, I.T., and Palma, M.
Arabian Journal of Chemistry (2021) 14, 103094

The growing demand for Spirulina (Arthrospira platensis) as a dietary supplement suggests that a rapid and reliable method to determine its saccharides and polyols content would be of great interest. Additionally, the impact of these microalga growing conditions may have on such content would be really useful for Spirulina producers. This manuscript presents the development and validation of a new analytical microwave-assisted extraction (MAE) method as the first step in determining saccharides and polyols in spirulina samples. A Box Behnken design has been employed to evaluate three extraction variables, viz., temperature (x1: 30, 55 and 80 C), solvent composition (x2: 0, 25, and 50% ethanol in water), and solvent to sample ratio (x3: 10:1, 20:1, and 30:1 mL of solvent per g of sample). These extraction variables have been optimized by Response Surface Methodology (RSM). Under the optimal conditions that have been thereby established (i.e., 30 C, 50% ethanol in water, and 30:1 solvent to sample ratio), a kinetic study has been performed with full recovery in 15 min. Subsequently, the developed MAE method was validated by determining the number of parameters, including linearity, LODs, LOQs, accuracy, and precision. Finally, the resulting MAE method was successfully applied to determine saccharides and polyols contents in a number of Spirulina samples that had been cultivated in varying salinity media. Five saccharides and polyols were identified in Spirulina, i.e., inositol, glycerol, sorbitol, glucose, and maltose. The total amount of the compounds of interest in the Spirulina that had been cultivated in a higher salinity media (17 and 25 g L1 of sodium chloride in water) was six-folds higher than the one cultivated in low saline water (0 and 3 g L1 of sodium chloride in water). This substantial content difference was mainly explained by the considerable increment in glycerol and glucose contents when grown in a more saline medium. Therefore, it has been demonstrated that the new method is suitable to determine saccharides and polyols contents in different Spirulina samples.

Methodologies in the Analysis of Phenolic Compounds in Roselle (Hibiscus sabdariffa L.): Composition, Biological Activity, and Beneficial Effects on Human Health

Journal Article
Hapsari, B.W., Manikharda, and Setyaningsih, W.
Horticulturae 2021, 7, 35.

Roselle (Hibiscus sabdariffa L.), as an edible flower, has long provided an array of positive effects on human health. This benefit is a result of phenolic compounds that are naturally present mainly in the calyx. Plentiful medicinal remedies and functional foods based on this flower are available worldwide, as supported by the studies of phenolic compounds in recent decades. This paper aims to provide a comprehensive review of the composition, biological activity, and beneficial effects on human health of phenolic compounds in roselle. This review was performed in accordance  with the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. A structured search in the published literature for phenolics compositions in roselle was required prior to the evaluation on the validity of the reported analytical methods. Reliable identification and quantification of phenolic compounds in roselle can be achieved by employing the proper extraction and separation methods. With ample alternative analytical methods discussed here, this review provided an aid for comprehending and selecting the most appropriate method for a particular study. The applications of the analytical methods highlighted indicated that phenolic acids, flavonoids, and their derivatives have been identified and quantified in roselle with a range of biological activities and beneficial effects on human health. It was also disclosed that the composition and concentration of phenolic compounds in roselle vary due to the growth factors, cultivars, and environmental influence. Finally, apart from the research progress carried out with roselle during the last ten years, this review also proposed relevant future works.

Oligo-glucomannan production from porang (Amorphophallus oncophyllus) glucomannan by enzymatic hydrolysis using β-mannase

Journal Article
Anggela, Setyaningsih, Wichienchot, S., W.. Harmayani, E.
Indonesian Food and Nutrition Progress 17(1):23-27

Porang (Amorphophallus oncophyllus) is an indigenous tuber of Indonesia that rich in glucomannan. An alternative approach to produce porang oligo-glucomannan (POG) as prebiotic from porang glucomannan (PGM) was made by enzymatic hydrolysis using β-mannanase. This study aimed to produce POG under optimal conditions by controlled enzymatic hydrolysis process. The PGM flour contained 96.12% of indigestible carbohydrates. The optimum condition of enzymatic hydrolysis producing the highest reducing sugar was as follows: temperature 37 °C, pH 5.5, a ratio of enzyme to the substrate (E/S) 1:1000, and reaction time 4 h. HPLC analysis confirmed that 99.45% of the resulting POG consisted of oligosaccharides with a degree of polymerization (DP) 3. Hence, the PGM utilized in this study has been proven as a potential substrate for POG production. Additionally, the resulting POG was considered as a functional ingredient due to has prebiotic potential.

A comprehensive study on the impact of the substituent on pKa of phenylboronic acid in aqueous and non-aqueous solutions: A computational approach

Journal Article
Kurnia, K.A., Setyaningsih, W., Darmawan, N., Yuliarto, B.
Journal of Molecular Liquids 326 (2021) 115321

The acid dissociation constant (pKa) is the fundamental physicochemical properties required to understand the structure and reactivity of boronic acid-based material as a sensor that identifies carbohydrates. However, there is a lack of comprehensive study on the impact of the substituent on the pKa of monosubstituted phenylboronic acid in aqueous and non-aqueous solutions. In this work, extensive experimental data on the pKa of monosubstituted phenylboronic acid in an aqueous solution was reviewed and compared in terms of accuracy. In addition, computational, were used to predict and investigate the impact of the substituent on the pKa for a series of monosubstituted phenylboronic acid in an aqueous solution at the molecular level. Good agreement was observed between predicted and literature pKa values of monosubstituted phenylboronic acid in the aqueous solution. While some deviations exist, predominantly with fluorine-containing phenylboronic acid, the COSMO-RS model is proficient at predicting the pKa of boronic acid in an aqueous solution with the accuracy of ±1.5 pKa. Subsequently, the model was used to predict the pKa of boronic acid in the non-aqueous solution, which data is not available in the literature. Furthermore, an excellent relationship is observed between the acidity of para-substituted, and to some extent, meta-substituted phenylboronic acid with the atomic charge of acidic hydrogen calculated using Natural Bond Orbital (NBO) Population Analysis. In contrast, the steric hindrance and the existence of other molecular forces might influence the acidity of ortho-substituted phenylboronic acid. The gathered information in this work could be of benefit for the understanding of the acidity of the boronic acid-based materials not only as a sensor but also in many diverse areas.

Process Optimization for Ultrasound-Assisted Starch Production from Cassava (Manihot esculenta Crantz) Using Response Surface Methodology

Journal Article
Setyaningsih, W., Karmila, Fathimah, R.N., Cahyanto, M.N
Agronomy 2021, 11(1), 117

Ultrasound-assisted extraction (UAE) has been optimized to improve the current cassava starch production by conventional maceration for the extraction method. Evaluation of several extraction parameters disclosed significant effects (p < 0.05) by three studied factors (ultrasound power, x1; pulse duty-cycle, x2; and solvent to sample ratio, x3). Subsequently, a Box-Behnken design (BBD) in conjunction with response surface methodology (RSM) was employed to optimise the three factors at three levels: x1 (30, 60, 90%), x2 (0.3, 0.6, 0.9 s1), and x3 (10:1, 20:1, 30:1). The model built for the RSM was validated through the coefficient of determination (R2 > 0.95), prediction error (2.12%), and lack-of-fit (0.71) values. The model validation suggested that the RSM was adequate for the observed data at the 95.0% confidence level. The optimum yield of cassava starch extraction was achieved by applying 90% for ultrasound power, pulse duty-cycle of 1.0 s1, and solvent to sample ratio of 30:1 with 10 min extraction time. Finally, the UAE produced starch with a purity of 88.36% and a lower viscosity than the commercial sample due to the granules’ size alteration. Hence, apart from speeding up the extraction process, UAE was worthwhile for the starch modification that could maintain the viscosity at a lower value (1920 cP) than the commercial starch (1996 cP) at the highest studied temperature treatment of 70 C.

Classification and prediction of patin fish oil’s antioxidant activities from different origins using FTIR spectroscopy and chemometrics

Journal Article
Putri, A.R., Rohman, A., Riyanto, S, and Setyaningsih, W.
International Journal of Pharmaceutical Research 13(1): 990–998

Patin fish oils (PFOs) from fifteen different origins was evaluated the antioxidant activity using 2,2’-diphenyl-1-picrylhydrazil (DPPH) radical scavenging. All PFO samples were analyzed using Fourier Transform Infrared (FTIR) spectroscopy. The spectra were recorded in absorbance mode and processed statistically using multivariate chemometric techniques, including Principal Component Analysis (PCA), Principal Component Regression (PCR) and Partial Least Squares (PLS). PCA was used to classify PFOs from different origins, while PLS and PCR was used to predict the antiradical activity of PFOs based on FTIR spectra. The result showed that radical scavenging activity (%RSA) of PFO from Demak and Boyolali (Central Java) had high value than others, 40.88 ± 0.18% and 40.71 ± 0.07%, respectively. PCA could successfully classify PFOs from fifteen different origins based on the absorbance of FTIR spectra. PCR using the first derivative spectra at 3005-650 cm-1 could predict %RSA of PFOs with R2 value of 0.9987 and 0.9982 in calibration and validation models, with RMSEC and RMSEP were 0.609 and 0.734, respectively.

Sida Rhombifolia Linn: Phytochemicals Composition and Biological Activities

Journal Article
Rohman, A., Ikhtiarini, A.N., Setyaningsih, W., Rafi, M., Aminah, N.S., Insanu, M., Irnawati, and Windarsih, A.
International Journal of Pharmaceutical Research 12(2): 2950-2958

The use of herbal medicine to treat several diseases has increased in the past few years. One of the plants that can be used for herbal medicine is Sida rhombifoila Linn either using whole part or leaves part. S. rhombifolia, known with Indonesian name as Sidaguri, belongs to the family of Malvaceae. The objective of this review was to highlight the chemical composition and biological activities of S. rhombifolia. During performing this review, several abstracts,  reports, and original articles were reviewed from several databases such as Scopus, Science direct, Springer, PubMed and Google Scholar. The keywords used were Sida rhombifolia or “Sidaguri”, phytochemical, pharmacological activities such as antioxidant, antidiabetic, anti-inflammatory, wound healing, analgesic, cytotoxic, antiproliferative, and anticholinesterase activity. S. rhombifolia Linn is very potential to be used as herbal medicine due to some active compounds contained in this plant including phenolic, flavonoid, saponin, and alkaloid. These compounds were responsible for its several pharmacological activities such as antidiabetic and hepatoprotective.

Review—Recent Trend on Two-Dimensional Metal-Organic Frameworks for Electrochemical Biosensor Application

Journal Article
Ulhakim, M.T., Rezki, M., Dewi, K.K., Abrori, S.A., Harimurti, S., Septiani, N.L.W., Kurnia, K.A., Setyaningsih, W., Darmawan, N., and Yuliarto, B.
Journal of The Electrochemical Society 167(13)

Electrochemical biosensors have attracted a tremendous attention for many researchers recently due to its facile synthesis process, tunability easiness by tailoring the material properties or composition, and wide range of biological analyte types detection. To obtain an excellent electrochemical biosensor performance, a material that facilitates fast electron transfer, large surface area, excellent electrocatalytic activity, and abundant available sites for bioconjugation is immensely needed. Metal-organic frameworks in the two-dimensional form (2D MOFs) provide all of the criteria needed as the sensing material for electrochemical biosensors application. However, the design and preparation of 2D MOFs, which have high stability and sensitivity as well as good selectivity for biological analyte detection, is still quite challenging. This review provides the recent studies and development of 2D MOFs as electrochemical biosensor. A detailed discussion about 2D MOFs structures, their synthesis strategy and control, 2D MOFs materials in electrochemical biosensor application, and the future challenges is thoroughly explained in this review. Hopefully, this review will also provide a new inspiration to advance future studies of 2D MOFs materials development as electrochemical biosensor.

Development of a methodology based on headspace gas chromatography-ion mobility spectrometry for the rapid detection and determination of patin fish oil adulterated with palm oil

Journal Article
Putri, A. R., Aliano-Gonzales, M. J., Ferreiro, M., Setyaningsih, W., Rohman, A., Riyanto, S., Palma, M.
Arabian Journal of Chemistry 13(10): 7524-7532.

The present research intends to develop a new method based on headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) for the detection and determination of patin fish oil adulterated with different percentages of palm oil, because it is a cheaper vegetable oil. Five variables that affect headspace generation (incubation time and temperature, amount of sample, injection volume, and pre-heating time) have been optimized by means of a Box Behnken design in combination with Response Surface Methodology. Then, repeatability and intermediate precision have been studied where coefficients of variation lower than 10% were obtained. This new method has been applied to several samples of patin fish oil adulterated with palm oil at 20 different levels (5–50% palm oil content). The results have confirmed the suitability of the HS-GC-IMS for a rapid, easy, and reliable detection and discrimination of adulterated oil samples since a characteristic fingerprint that allows 100% successful discrimination between adulterated and unadulterated patin oil samples was achieved. Finally, a regression model has been developed to determine each sample’s adulteration level with an error lower than 10% and a coefficient of determination greater than 0.95.

P

Development and Validation of HPLC-DAD Method for Simultaneous Determination of Seven Food Additives and Caffeine in Powdered Drinks

Journal Article
Imanulkhan, Setyaningsih, W., Rohman, A., and Palma, M.
Foods 9(1119)

The usage of food additives must respect the general legislation in force in the country and requires a reliable analytical method for surveillance. This research aimed to develop a high-performance liquid chromatography with diode array detection (HPLC-DAD) method for the
simultaneous determination of seven food additives and ca
ffeine in powdered drinks. Three factors likely to affect the chromatographic separation, namely, mobile phase composition at the beginning (x1, 0–10% of the amount of methanol in the phosphate buffer) and the end (x2, 60–100% of the amount of methanol in the phosphate buffer) of the gradient program and pH (x3, 3–7), were evaluated with the aid of a Box–Behnken Design (BBD). Subsequently, multi-response optimizations for chromatographic resolutions (Rs) and analysis time were performed using the response surface methodology (RSM) in conjunction with the desirability function (DF). Complete separation (Rs > 1.5) of seven food additives and caffeine was achieved in less than 16 min by applying 8.5% methanol in the phosphate buffer at the beginning and 90% at the end of the gradient program, in pH 6.7. The developed method was validated with low limits of detection (ranging from 1.16 mg kg1 (sodium saccharin) to 3.00 mg kg1 (acesulfame potassium)), low limits of quantification (ranging from 3.86 mg kg1 (sodium saccharin) to 10.02 mg kg1 (acesulfame potassium)), high precision (CV < 4%), and high accuracy (recoveries from 95 to 101% at 80, 100, and 120% of the target concentration). The method was successfully used to assess the seven food additives and caffeine in commercially available powdered drinks.

foods Pak Iman

Application of FTIR Spectroscopy and HPLC Combined with Multivariate Calibration for Analysis of Xanthones in Mangosteen Extracts

Journal Article
Tejamukti, E.P., Setyaningsih, W., Irnawati, Yasir, B., Alam, G., Rohman, A.
Scientia Pharmaceutica 88(35)

Mangosteen, or Garcinia mangostana L., has merged as an emerging fruit to be investigated
due to its active compounds, especially xanthone derivatives such as
α -mangostin (AM), γ-mangostin (GM), and gartanin (GT). These compounds had been reported to exert some pharmacological activities, such as antioxidant and anti-inflammatory, therefore, the development of an analytical method capable of quantifying these compounds should be investigated. The aim of this study was to determine the correlation between FTIR spectra and HPLC chromatogram, combined with chemometrics for quantitative analysis of ethanolic extract of mangosteen. The ethanolic extract of mangosteen pericarp was prepared using the maceration technique, and the obtained extract was subjected to measurement using instruments of FTIR spectrophotometer at wavenumbers of 4000–650 cm1 and HPLC, using a PDA detector at 281 nm. The data acquired were subjected to chemometrics analysis of partial least square (PLS) and principal component regression (PCR). The result showed that the wavenumber regions of 3700–2700 cm1 offered a reliable method for quantitative analysis of GM with coefficient of determination (R2) 0.9573 in calibration and 0.8134 in validation models, along with RMSEC value of 0.0487% and RMSEP value 0.120%. FTIR spectra using the second derivatives at wavenumber 3700–663 cm1 with coefficient of determination (R2) >0.99 in calibration and validation models, along with the lowest RMSEC value and RMSEP value, were used for quantitative analysis of GT and AM, respectively. It can be concluded that FTIR spectra combined with multivariate are accurate and precise for the analysis of xanthones.

Tejamukti FTIR

Banana Peel Flakes Alleviate Blood Glucose and Stress in A Dose-Dependent Manner

Journal Article
Meliala, A., Sumiwi, Y.A.A., Narwidina, P., Rini, S.L.S., Setyaningsih, W.
Int J Pharm Pharm Sci 12(8): 75-81

Objective: This study aimed to evaluate the antidiabetic and antidepressant effects of banana peel flakes in streptozotocin-induced diabetic rats.
Methods: Twenty-five male Wistar rats were classified into five groups with different treatments. Groups I to IV were diabetic rats model groupsthat consumed only standard diet, standard diet containing 5%, 10%, and 20% of banana peel flakes, respectively. While group V was a healthy
control group fed a standard diet. Immunohistochemistry staining was measured to examine serotonin expression in the colon and pancreas.
Results: The diabetic rats treated with 20% banana peel flakes had a lower blood glucose concentration (p<0.05) compared with diabetic control and showed a shorter duration of immobility time (p<0.05) than the healthy control. Additionally, compared with diabetic control, the diabetic rats treated with 5% banana peel flakes showed higher serotonin expression (p<0.05) in the colon. In contrast, serotonin expression in the pancreas did not show any significant difference (p>0.05).
Conclusion: The present study disclosed that the banana peel flakes provided an antidepressant effect in the diabetic rats model, which might occur through the mechanism of controlling blood glucose concentration.

 

Banana Peels

Determination of acid, peroxide, and saponification value in patin fish oil by FTIR spectroscopy combined with chemometrics

Journal Article
Putri, A.R., Rohman, A., Setyaningsih, W. and Riyanto, S.
Food Research 4 (5): 1758 - 1766

Simple, rapid, and reproducible methods for determining the acid value (AV), peroxide value (PV), and saponification value (SV) of patin fish oil (PFO) were developed using Fourier Transform Infrared (FTIR) spectroscopy combined with chemometrics of Principal Component Regression (PCR) and Partial Least Square (PLS). The relationship between actual values was determined using AOCS method and predicted value was determined with FTIR spectroscopy and chemometrics. From the validation work, the high coefficient of determination (R2) reached up to > 0.99. This study concluded that by means of FTIR spectra that combined with PCR and PLS technique can be used to determine AV, PV, and SV of PFO.

Patin Fish Oil Food Research

Effect of the harvesting period on the phenolic content and on antioxidant activity of two Algerian olive cultivars

Journal Article
Deflaoui, L., Mettouchi, S., Setyaningsih, W., Lovillo, MP., Barroso, CG., and Tamendjari, A.
La Rivista Italiana Delle sostanze Grasse XCVII: 51-60

Because of their antioxidant and anti-free radical capacities, the phenolic compounds of olives gained growing interest.The aim of this work is to determine the harvest optimal period of two Algerian olive varieties (Chemlal and Blanquette de Guelma) harvested at four stages of maturity which will allow the best equilibrium between the olives’ composition in phenolic compounds and their antioxidant activities.
Phenolic compounds were determined by colorimetric methods and by UPLC-PDA. The antioxidant activity of olive extracts was assessed by measuring the reducing power, the antiradical activity toward DPPH radical andABTS.
Results confirm that the process of maturation and the variety of olives affect considerably the phenolic compounds and their antioxidant activity. As a consequence, a significant decrease in the antioxidant activity (reducing power and antiradical activities) of the olive fruits is observed. The relative antioxidant capacity index (RACI), which represents the mean scores of the antioxidant activities, showed that the highest indices were obtained by the green stage for
Chemlal and spotted green stage for Blanquette de Guelma.
The harvesting can occur when the olives are in the spotted green stage, corresponding to the middle of December. This period allows a good combination between the phenolic content and antioxidant activity.

Capture

Consumers’ willingness to pay for functional rice: a survey from Indonesia

Journal Article
Annur, N.D., Nugrohoningtyas, B.S.H., Rodríguez Dodero, M.C. and Setyaningsih, W.
Food Research 4 (4) : 1344 - 1350

Rice, especially the pigmented varieties, contains a higher level of melatonin (N-acetyl-5-methoxytryptamine), which is used in treatment for sleeping disorders and for improving mood. It has potent antioxidant properties, anti-obesity, and anti-inflammatory, thus potential as the basis for functional food development. The purpose of the study was to examine how the various rice attributes and demographic characteristics affect the willingness to pay of Indonesian consumers on functional rice. Multi-Factor Categorical Design (MFCD) and Balanced Incomplete Block Design (BIBD) have been used to study multiple non-quantitative factors, including different rice varieties and health benefits information on the label (x1, 3 levels: white rice, brown rice-label, and red rice-label), price (x2, 2 levels: low and high), and rice producers (x3, 3 levels: international brand, established national brand, and new national brand). Data collection was administered through an online questionnaire survey to 204 respondents, living in four major Indonesian islands: Java, Bali, Sumatra, and the Moluccas. The results showed that despite the health benefits of the melatonin contained in the whole brown and red rice, Indonesian consumers still preferred conventional white rice over the aforementioned rice varieties. Price was recognized as the main attribute affecting the selection of functional rice, whilst rice producers gave less impact on willingness to pay. The results indicated that cultural value, consumers’ knowledge of melatonin, consumers’ health status, and consumers’ income were the sources of heterogeneity in consumers’ willingness to pay for functional rice.

Untitled

Valorization of Jack Bean as Raw Material for Indonesian Traditional Food Tempeh and Its Functional Properties

Journal Article
Andriati Ningrum , Sri Anggrahini and Widiastuti Setyaningsih
Journal of Applied Sciences, 19(2): 56-61.

Jack bean is one of underutilized legume. In Indonesia jack bean tempeh is one traditional product from Indonesia, using jack bean tempeh as food ingredient. So far, scientific information on jack bean tempeh is scarce and thus this article aims to document the updated knowledge of jack bean tempeh. Tempeh is a sliceable, cake-like product made of dehulled cooked jack bean, penetrated and fermented by a mixed microbiota dominated by filamentous fungi. Jack bean as one type of pulses are a rich source of protein in the human diet and their consumption has been associated with the prevention of chronic diseases. The beneficial effect in human health has been related to their micronutrients, phytochemical bioactive compounds and recently BP (bioactive peptides).

Untitled1

Assessment of Microbiological Quality and Mycotoxin in Dried Chili by Morphological Identification, Molecular Detection, and Chromatography Analysis.

Journal Article
Wikandari, R., Mayningsih, I.C., Sari, M.D.P., Purwandari, F.A., Setyaningsih, W., Rahayu, E.S., Taherzadeh, M.J..
International Journal of Environmental Research and Public Health 17 (6) 1847

The growing interest in spicy foods leads to the global demand for spices, particularly dried chili. This study aimed to assay both aflatoxin (AFs) and ochratoxin A (OTA) contamination using an integrative method of morphological identification, molecular detection, and chromatography analysis on dried chili provided from traditional and modern markets in Indonesia. The results showed that total fungal infection ranged from 1-408 × 103 CFU/g. Eighty percent of the chili obtained from both the traditional and the modern markets were infected by Aspergillus spp., in which 50% of the infections were identified as A. parasiticus and A. flavus. A complete set of targeted genes involved in AF production and OTA were detected in two isolates of A. flavus and one isolate of A. carbonarius, respectively. The levels of AFs B1, B2, and OTA in the contaminated dried chilies were in the range of 39.3–139.5 µg/kg, 2.6–33.3 µg/kg, and 23.7–84.6 µg/kg, respectively. In contrast, no AFs G1 and G2 were detected. This study showed that the fungal infection of Indonesian dried chili occurs both in the field and during storage; thus, it is suggested to implement good agricultural and handling processes.

Untitled

Physicochemical properties of oci fish joruk (Rastrelliger kanagurta) and the antioxidant activity of joruk hydrolysate

Journal Article
Anggrahini, S., Setyaningsih, W., Ningrum, A., Anto, Agustiari, N.M.
Food Research 4(3), pp. 786-792

Joruk is a traditional fishery product made by spontaneous fermentation process using lactic acid bacteria by salting method, followed by fermentation process. Hydrolysis of proteins with the pepsin and a combination of pepsin and trypsin enzymes by in vitro can produce bioactive peptides. The aims of this study were to investigate the change of soluble protein and lactic acid content in oci fish joruk (Rastrelliger kanagurta) and to determine the influence of the enzyme hydrolysis on the antioxidant activity of joruk. The raw material used in this highest study was the oci fish obtained from Gorontalo, Indonesia. The produced joruk was added with the concentration of palm sugar 10%, 20% and 30% then fermented for 8 days, 10 days and 12 days. Protein hydrolysis using the pepsin enzyme and a combination of pepsin and trypsin enzyme. The results showed that the addition of palm sugar and fermentation time can reduce soluble protein and increase the lactic acid content. The highest antioxidant activity was found in joruk hydrolysate that hydrolyzed by pepsin enzyme with the addition of 30% palm sugar and 12 days fermentation, with a value of 5.28%.

Untitled

Key-marker volatile compounds in aromatic rice (Oryza sativa) grains: An HS-SPME extraction method combined with GC×GC-TOFMS

Journal Article
Setyaningsih, W., Majchrzak, T., Dymerski, T., Namieśnik, J., Palma, M.
Molecules, 24(22), 4180

The aroma of rice essentially contributes to the quality of rice grains. For some varieties, their aroma properties really drive consumer preferences. In this paper, using a dynamic headspace solid-phase microextraction (HS-SPME) system coupled to a two-dimensional gas chromatography (GC ×GC) using a time-of-flight mass spectrometric detector (TOFMS) and multivariate analysis, the volatile compounds of aromatic and non-aromatic rice grains were contrasted to define some chemical markers. Fifty-one volatile compounds were selected for principal component analysis resulting in eight key-marker volatile compounds (i.e., pentanal, hexanal, 2-pentyl-furan, 2,4-nonadienal, pyridine, 1-octen-3-ol and (E)-2-octenal) as responsible for the differences between aromatic and non-aromatic rice varieties. The factors that are most likely to affect the HS-SPME efficiency for the aforementioned key-marker compounds were evaluated using a 25III−2 fractional factorial design in conjunction with multi-response optimisation. The method precision values, expressed as % of coefficient of variation (CV), were ranging from 1.91% to 26.90% for repeatability (n = 9) and 7.32% to 37.36% for intermediate precision (n = 3 × 3). Furthermore, the method was successfully applied to evaluate the volatile compounds of rice varieties from some Asian countries.

Screenshot_8

Optimisation of an ultrasound-assisted extraction method for the simultaneous determination of phenolics in rice grains

Journal Article
Setyaningsih, W., Saputro, I.E., Carrera, C.A., Palma, M.
Food Chemistry Volume 288, 1 August 2019, Pages 221-227

Fifteen phenolic compounds were determined in rice grains by ultrasound assisted extraction and ultra-performance liquid chromatography with photodiode array detection. Primarily, an UAE method has been developed and validated for the extraction of phenolics from rice grains. For the optimization, a Box–Behnken Design based on six factors including extraction temperature (10–70 °C), solvent composition (0–50% methanol in water), cycle (0.2–0.7 s−1), ultrasound amplitude (30–70%), and solvent to sample ratio (2.5:1 to 5:1) was employed. Multi-response optimization (MRO) was performed to develop a simultaneous extraction method of 15 phenolics. The optimal UAE conditions were: extraction temperature 45 °C, time 25 min, cycle 0.4 s−1, ultrasound amplitude 47%, solvent composition 80% methanol in water at pH 4.25, and sample to solvent ratio 1:5. Linearity, LODs, LOQs, precision and recovery were determined. In addition, the method was successfully applied to analyse a number of rice samples with different varieties.

Screenshot_7

Glycerolysis-interesterification of chicken-palm stearin blend: Effect of solvent and chicken stearin to palm stearin ratio

Conference Paper
Sari, D.A., Ningsih, D.S., Nusantoro, B.P., Setyaningsih, W., Hidayat, C.

Both glycerolysis and the interesterification reactions take place in one reaction system. The objectives of this research were to obtain chicken stearin and to evaluate the effect of solvent to fat ratios and chicken stearin to palm stearin ratios on total mono-acylglycerol (MAG) and diacylglycerol (DAG) of product by the chemical glycerolysis-interesterification (GIE). GIE was initiated by adding NaOH to the reactant. Structured Lipids were evaluated based on acylglycerol composition and melting behavior. Results show that the yield of chicken stearin was 13%. The highest total MAG and DAG was obtained at the stearin: solvent ratio 1:3 (w/v). An increase in chicken fat stearin to palm stearin caused a slight increase in the total MAG and DAG. However, it didn’t have a significant increase in the total MAG and DAG. The best ratio of palm stearin to chicken stearin was 40:60 because slip melting point and melting point were 36.16 ± 4.9°C and 37.59 ± 5.96°C. Thus, the best condition was obtained at the ratio of stearin to solvent 1: 3 (w/v), temperature 50°C, oil to glycerol ratio 1:5, 300 rpm and NaOH 3%. The reaction was performed for 2 h. Total MAG and DAG was 67.87 ± 1.97%.
Screenshot_6

Influence of palm sugar to the functional properties of bekasam hydrolysate

Journal Article
Anggrahini, S., Setyaningsih, W., Ningrum, A., Anto, Agustiari, N.M.
Food Research 3 (3) : 265 -271

Hypertension is one of the major global health issues worldwide. One of the methods can be used to treat this disease is by inhibiting the activity of angiotensin converting enzyme, an enzyme that controls the regulation of arterial blood pressure and cardiac output. Bekasam is one of the traditional fermented food made from freshwater fish (oci, Rastreliger kanaguria) with the addition of carbohydrate sources (variation concentration of palm sugar i.e. 10%; 20%; and 30%). The objective of this research was to investigate the effect of palm sugar concentration to the ACE inhibitor activity of bekasam after digestive enzyme degradation by trypsin. Hydrolysis process was evaluated by determination of the concentration of soluble protein and the degree of hydrolysis (DH) after treatmentand the ACE-inhibitory (ACEI) effect was determined by spectrophotometry method. Compared to the control without trypsin treatment (5.00±0.10 mg/ml), theconcentration of soluble protein (mg/ml)of bekasam wasincreased after trypsin hydrolysis to 16.57±0.55 at 10% of added palm sugar and then decreased to 15.38±0.18 and 10.85±0.28, at 20 and 30% of added palm sugar respectively. The degree of hydrolysis was 21.3±0.3%, 19.7±0.5% and 15.3±0.4%, for 10, 20 and 30% of palm sugar, respectively. The ACE-inhibitory activity was 87.39±0.01, 84.52±0.84, and 77.68±0.63%, for 10, 20 and 30% of added palm sugar, respectively. The results showed that the concentration of palm sugar affects the soluble protein content, degree of hydrolysis and ACEI activity after trypsin hydrolysis with the optimum sugar concentration was 10%.

Screenshot_5

Fast Determination of Phenolic Compounds in Rice Grains by Ultraperformance Liquid Chromatography Coupled to Photodiode Array Detection: Method Development and Validation

Journal Article
Setyaningsih, W., Saputro, I.E., Carrera, C.A., Palma, M., García-Barroso, C.
J. Agric. Food Chem.2019, 67, 3018−3027

There are several phenolic compounds in rice grains providing benefits for human health. The concentration of phenolic compounds in rice is strongly affected by the polishing steps during rice production. A new sensitive ultra performance liquid chromatography−ultraviolet−visible spectroscopy method with a photodiode array detection protocol has been developed and validated for the quantitation of phenolic compounds in rice grains. Several working variables and two different columns were evaluated. Finally, a less than 3 min analysis time was developed to achieve enough resolution for the simultaneous determination of the 20 most common phenolic compounds in rice. The analytical properties for the separation method produced an adequate sensitivity for all phenolic compounds in the regular range for phenolics in rice, 0.5−100 mg L−1(R2> 0.997), with high precisions for both repeatability and intermediate precisions (coefficients of variation less than 0.4 and2.5% for the retention time and area of the peaks, respectively).Screenshot_4

Physicochemical characterization of jack bean (Canavalia ensiformis) tempeh

Journal Article
Andriati, N., Anggrahini, S., Setyaningsih, W., Sofiana, I., Pusparasi, D.A., Mossberg, F.
Food Research 2 (5) : 481 -485

The objective of our research was to investigate the properties of jack bean tempeh during fermentation with two different packaging types, banana leaves and LDPE (Low-Density Poly Ethylene) Plastic. After 24 hrs mycelia of Rhizopushasnot been formed, but it fully formed after 36 hrs fermentation for all formulated tempeh with two different packaging. The mycelia started the change to form brownish colour after 36 hrs of fermentation. Solid state fermentation (SSF) for 0, 24, 36, 48 and 60 h showed influence in different parameter e.g. total protein, the colour parameter (L, a, b, C, Hue, and ΔE) and pH during fermentation (p<0.05). Optimum condition of tempeh was chosen after 36 hrs fermentation that showed the influence of different packaging type to the lipid content and antioxidant activity (p<0.05), but no significant different to the water content, ash content, protein content, dietary fiber content and carbohydrate by the different content of tempeh (p>0.05). In the present research work, we demonstrated that different fermentation time and packaging will give influence to several physiochemical characteristics including total soluble protein, colour, and pH.

Screenshot_3

The adsorptive properties of UiO-66 towards organic dyes: A record adsorption capacity for the anionic dye Alizarin Red S.

Journal Article
Embaby, M.S., Elwany, S.D., Setyaningsih, W., Saber, M.R.
Chinese Journal of Chemical Engineering, Volume 26, Issue 4, April 2018, Pages 731-739

In order to decisively determine the adsorption selectivity of zirconium MOF (UiO-66) towards anionic versus cationic species, the adsorptive removal of the anionic dyes (Alizarin Red S. (ARS), Eosin (E), Fuchsin Acid (FA) and Methyl Orange (MO)) and the cationic dyes (Neutral Red (NR), Fuchsin Basic (FB), Methylene Blue (MB), and Safranine T (ST)) has been evaluated. The results clearly reveal a significant selectivity towards anionic dyes. Such an observation agrees with a plethora of reports of UiO-66 superior affinity towards other anionic species (Floride, PO43 −, Diclofenac sodium, Methylchlorophenoxy-propionic acid, Phenols, CrO42 −, SeO32 −, and AsO4). The adsorption process of ARS as an example has been optimized using the central composite design (CCD). The resultant statistical model indicates a crucial effect of both pH and sorbent mass. The optimum conditions were determined to be initial dye concentration 11.82 mg.L− 1, adsorbent amount 0.0248 g, shaking time of 36 min and pH 2. The adsorption process proceeds via pseudo-second order kinetics (R2 = 0.999). The equilibrium data were fit to Langmuir and Tempkin models (R2 = 0.999 and 0.997 respectively). The results reveal an exceptional removal for the anionic dye (Alizarin Red S.) with a record adsorption capacity of 400 mg·g− 1. The significantly high adsorption capacity of UiO-66 towards ARS adds further evidence to the recently reported exceptional performance of MOFs in pollutants removal from water.

Screenshot_2

Optimization of the ultrasound-assisted extraction of tryptophan and its derivatives from rice (Oryza sativa) grains through a response surface methodology

Journal Article
Setyaningsih, W., Saputro, I.E., Palma, M., Barroso, C.G.
Journal of Cereal Science, 2017, 75: 192-197, DOI: 10.1016/j.jcs.2017.04.006.

ABSTRACT

An analytical ultrasound-assisted extraction (UAE) technique has been optimized and validated for the extraction of tryptophan and its derivatives from rice grains. A Box–Behnken design in conjunction with a response surface methodology based on six factors and three levels was used to evaluate the effects of the studied factors prior to optimizing the UAE conditions. The significant (p < 0.05) response surface models with high coefficients of determination were fitted to the experimental data. The most significant (p < 0.0001) effect is the solvent-to-sample ratio while quadratic effects caused by temperature and solvent-to-sample ratio were of moderate importance (p < 0.05). The optimal UAE conditions were as follows: extraction time of 5 min, ultrasound amplitude of 30%, cycle of 0.7 s−1, extraction temperature of 30 °C, 8% methanol in water as the extraction solvent at pH 3 and a solvent/solid ratio 5:1. The method validation ensured that appropriate values were obtained for the LOD, LOQ, precision and recovery. Furthermore, the method was successfully applied to the analysis of a number of rice samples of different varieties. It was demonstrated that this particular UAE method is an interesting tool for the determination of tryptophan and tryptophan derivatives in rice grain samples.

JCS_02

 

Tryptophan Levels during Grape Ripening: Effects of Cultural Practices

Journal Article
Ruiz-Rodríguez , A., Carrera, C.A., Setyaningsih, W., Barbero, G.F., Ferreiro-González, M., Palma, M., Barroso, C.G.
Molecules, 2017, 22(6), 941: 1-9, DOI:10.3390/molecules22060941

ABSTRACT

Some cultural practices that are carried out during the grape ripening period are associated with vine stress, including leaf removal, grape bunch removal, and vegetable cover crops. Additionally, several nitrogen and sulfur supplements have also been used directly on leaves during the last stage of the ripening period. In the work described here, five different cultural practices and the reference were applied in three replicates in the same vineyard. The evolution of tryptophan levels was evaluated from just after grape veraison until the harvest date. In some cases, certain specific treatments were also evaluated after the regular harvest date. The cultural techniques that involved the application of nitrogen led to higher levels of tryptophan at the harvest day when compared to other cultural techniques. It was also found that the application of nitrogen without sulfur had a faster effect on the level of tryptophan. It was established that a period of around 20 days is needed for the grapes to show clear differences in tryptophan levels after the application of nitrogen.

Molecules

Profile of phenolic compounds in Indonesian rice (Oryza sativa) varieties throughout post-harvest practices

Journal Article
Setyaningsih, W., Hidayah, N., Saputro, I. E., Palma, M., Barroso, C. G
Journal of Food Composition and Analysis, 2016, 54: 55–62, DOI: 10.1016/j.jfca.2016.10.004

Abstract

Apart from being a foremost source of calories for a large part of the world’s population, cultivated rice (Oryza sativa) provides beneficial effects for human health through several bioactive compounds, including phenolics. Therefore, to earn this advantage, it is essential to retain the phenolics through appropriate post-harvest practices. In this paper, the profile of 10 individual phenolic compounds contained by rice grains after drying, dehusking and polishing processes were studied. A number of Indonesian rice cultivars from indica and javanica subspecies were acquired from conventional (IR-64, Umbul-umbul and Pandanwangi) and organic farming (Batang Lembang, Pandanwangi, black- and red-pigmented rice) were studied. Additionally, the effect of polishing degree on individual phenolics content in the grain was also evaluated. The level of phenolics decreased, also their relative values dramatically changed throughout a series of post-harvest processes. Moreover, changes on the composition of phenolic acids and its aldehydes in the grain have been described. p-Coumaric acid reflects the greatest differences on the phenolic composition during the course of post-harvest practices that altered from 48% to 8% portion to the total phenolics in paddies and polished grains, respectively. However, phenolics content in black-pigmented rice appeared to be the most persistent in the rice matrix. Since the profile of individual phenolics has also been evaluated on different polishing degrees (70%, 80%, 90% and 100% bran removal), the distribution of these compounds in the whole grain rice was effectively disclosed. These variations in the compositions and quantities of phenolics revealed the effect of post-harvest practices, varietal differences as their distribution in the grain.

Artikel PHE post harvest

Multiresponse optimization of a UPLC method for the simultaneous determination of tryptophan and 15 tryptophan-derived compounds using a Box-Behnken design with a desirability function

Journal Article
Setyaningsih, W., Saputro, I.E., Carrera, C.A., Palma, M. , Barroso, C.G.
Food Chemistry, 2017, 225: 1-9, DOI: 10.1016/j.foodchem.2016.12.034

Abstract

A Box–Behnken design was used in conjunction with multiresponse optimization based on the desirability function to carry out the simultaneous separation of tryptophan and 15 derivatives by Ultra Performance Liquid Chromatography. The gradient composition of the mobile phase and the flow rate were optimized with respect to the resolution of severely overlapping chromatographic peaks and the total run time. Two different stationary phases were evaluated (hybrid silica and a solid-core-based C18 column). The methods were validated and a suitable sensitivity was found for all compounds in the concentration range 1–100 μg L–1 (R2 > 0.999). High levels of repeatability and intermediate precision (CV less than 0.25% and 1.7% on average for the retention time and the signal area, respectively) were obtained. The new method was applied to the determination tryptophan and its derivatives in black pigmented glutinous and non-glutinous rice grain samples.

Article 2017

Use of multiple response optimization for the separation of tryptophan and 15 related compounds by ultrahigh performance liquid chromatography

Conference Paper
Saputro I.E., Setyaningsih, W., Carrera, C., Palma, M., and Barroso, C.G.
The 14 international Congress on Amino Acids, Peptides and Proteins (Amino Acids 2015), August 37, 2015, Vienna, Austria.

Abstract

A Box Behnken Design (BBD) in conjunction with multiple response optimizations was performed to develop the simultaneous separation of tryptophan and fifteen related compounds in rice grains by Ultrahigh Performance Liquid Chromatography (UPLC) coupled with Photodiode Array (PDA) detector. The gradient compositions of mobile phase (%B start and %B end) and flow rate were optimized with respect to the resolutions of severely overlapping chromatographic peaks between the phenolic compounds in addition to the total analysis time. Furthermore, two different stationary phases were also evaluated (Cortex C18 and BEH C18). A gradient program of only 4 min analysis run using Cortex C18 was found to provide the best resolution for the simultaneous separation of the 16 studied compounds. The method was validated resulting an adequate sensitivity in the range of 1–100 μg L-1 (R2=0.9993-0.9999) with high precisions (CV lower than 2% in average for both repeatability and intermediate precisions). The suitability of the method was checked by application to the analysis of pigmented and non-pigmented rice samples. Tryptophan and several related compounds were identified and quantified in the tested samples.

Effects of Cultivar Practices on Tryptophan Levels during Grape Ripening

Conference Paper
Ruiz-Rodríguez, A., Setyaningsih, W., Carrera, C., Palma, M., and Barroso, C.G.
The 14 international Congress on Amino Acids, Peptides and Proteins (Amino Acids 2015), August 37, 2015, Vienna, Austria.

Abstract

Different cultivar practices have been applied to vine to promote specific components related to final wine flavor. Some of the cultivar practices are related to vine stress, including leaf removal, grape bunch removal and vegetable cover crops. Some nitrogen and sulfur supplements have been also used directly to leaves during the last stage of the ripening period. Main effects of cultivar practices are related to the final composition of organic acids, volatiles components and phenolics. However some effects on amino acids have been also found.

This paper specifically deals with the effects on tryptophan level because of it has strong effects in the evolution of several other related compounds, including auxins and melatonin. Six different cultivar practices were applied in three replicated in the same vineyard. Evolution of tryptophan levels were determined starting just after grape veraison till harvest date. In some cases, some specific treatments were also evaluated after regular harvest.

Results point out the specific effect of vegetable cover crops and leaves removal to promote higher levels of tryptophan in grapes.

Ultrasound-assisted Extraction (UAE) for Tryptophan and lts Derivatives from Rice (Oryza sativa) grains

Conference Paper
Saputro I.E., Setyaningsih, W., Carrera, C., Palma, M., and Barroso, C.G.
The 14 international Congress on Amino Acids, Peptides and Proteins (Amino Acids 2015), August 37, 2015, Vienna, Austria.

Abstract

Cultivated rice (Oryza sativa) is a global staple food that subsists calories for more than half of the world’s population. Emerging evidence supports that a number of bioactive compounds have been identified in rice including tryptophan and its derivatives (oxitriptan, serotonin, tryptamine and auxin). The development of single optimized extraction of bioactive compounds from plant may complicate due to the structural diversity. A new ultrasound-assisted extraction (UAE) of tryptophan and its derivatives has been optimized and validated.Experiment with a three-level and six-factor Box-Behnken Design (BBD) combined with Response Surface Methodology (RSM) was employed to evaluate the significances of studied factors (solvent composition,temperature, solvent-solid ratio, pH, amplitude and cycle) on the extraction yields. Subsequently, extraction kinetic was studied to confirm satisfactory recovery of the extracted compounds from the matrix. Later, by chromatographic analyses, tryptophan, oxitriptan, serotonin, tryptamine and 3-indole acetic acid (auxin) were identified and quantified in the extracts of rice grains.

A complete validation of the method was performed and presented agreeable linearity (correlation coefficients higher than 0.9998) with high extraction precisions (in average, expressed as CV, repeatability and intermediate precision were lower than 2.0% and 2.6% respectively). The method was successfully applied in the extraction of a number of real samples. Hence, it can be concluded that UAE under optimum conditions can be considered as a powerful tool for the extraction of tryptophan and its derivatives from a wide variety of rice grains.

Optimisation of Ultrasound-assisted Extraction (UAE) for Phenolics from Rice (Oryza sativa) Grains using Box-Behnken Design

Conference Paper
Setyaningsih, W., Saputro, I.E., Palma, M., and Barroso, C.G.
The GDCh Scientific Forum Chemistry (Chemie 2015), August 30-September 2, 2015, Dresden, Germany.

Abstract

Emerging evidence supports that a number of health-promoting substances have been identified in rice (Oryza sativa) including phenolic compounds. The development of single optimized extraction of phenolic compounds from plant may complicate due to the structural diversity as having multiple hydroxyl groups which results the polarities of these compounds vary significantly. Therefore, an optimised extraction method that effectively extracts phenolics from rice is needed prior to the reliable determination of these compounds in the grains.

A new ultrasound-assisted extraction (UAE) of phenolics has been optimized and validated. A three-level, six-factor Box-Behnken design in conjunction with Multi-responses optimisation was employed to evaluate the significances of studied factors (solvent composition, temperature, solvent-solid ratio, pH, amplitude and cycle) on the extraction yields. Subsequently, extraction kinetic was studied to confirm high recovery of phenolics extracted from the matrix. The levels of phenolics were quantified by Ultrahigh Performance Liquid Chromatography coupled with Photo-diode Array detector (UPLC-PDA) in less than 5 min.

The optimal UAE condition was as follows: extraction time 25 min, ultrasound amplitude 47%, cycle 0.4 s-1, extraction temperature 45oC, pH 5.5, extraction solvent 80% MeOH in water and solvent-solid ratio 5:1. A complete validation of the method was performed and presented agreeable linearity (correlation coefficients between 0.998 and 0.999) with high precision (in average, expressed as CV, repeatability and intermediate precision were lower than 3 and 4.5% respectively). The method was successfully applied in the extraction of a number of real samples in order to evaluate the applicability. Hence, it can be concluded that UAE under optimum conditions can be considered as a powerful tool for the determination of phenolic compounds from a wide variety of rice grains.

Multiresponse HS-SPME Optimization Combined with GC×GC-TOFMS for Key-marker Aroma Compounds in Indonesian Aromatic Rice (Oryza sativa)

Conference Paper
Setyaningsih, W., Majchrzak, T., Dymerski, T., Namieśnik, J., Palma, M., Barroso, C. G.
The GDCh Scientific Forum Chemistry (Chemie 2015), August 30-September 2, 2015, Dresden, Germany.

Abstract

The grain aroma is a primary sensory attribute of high-quality rice that contributes a critical impact on consumer preference. Aromatic rice possesses typical volatile compounds released from the grain that discriminates aromatic rice varieties from ordinary ones. These varieties have become more widely appreciated in the current market. Furthermore, recent researches have led to an increase in rice breeding programs focusing on the odour profile to generate high quality aromatic rice cultivars. Therefore, an analytical method for key-marker aroma compounds determination is crucial to facilitate the selection of lines with superior aroma attributes.Using a dynamic HS-SPME system coupled to a GC×GC-TOFMS and multivariate analysis, the aroma chemistry of Indonesian aromatic and non-aromatic rice grains were compared and characterised to define the chemical markers of aroma quality. Sequentially, the analytical method of HS-SPME tandem with GC×GC-TOFMS for the determination of key marker aroma compounds in rice grains were developed and validated. The factors that are most likely to affect the HS-SPME efficiency were evaluated using a 2_III^(5-2) fractional factorial design in conjunction with multi-response optimisation.

Separation of 20 Phenolic Compounds in Rice by Ultrahigh Performance Liquid Chromatography (UPLC)

Conference Paper
Setyaningsih, W., Saputro, I.E., Barbero, G., Palma, M., and Barroso, C.G.
The 18 Scientific Meeting of the Euroanalysis series (EuroAnalysis 2015), September 6-10, 2015, Bordeaux, France

Abstract

A Box Behnken Design (BBD) in conjunction with multiple response optimizations was performed to develop the simultaneous separation of twenty phenolic compounds in rice grains by Ultrahigh Performance Liquid Chromatography (UPLC) coupled with Photodiode Array (PDA) detector. The gradient compositions of mobile phase and flow rate were optimized with respect to the resolutions of severely overlapping chromatographic peaks between the phenolic compounds and the analysis time. Additionally, two different stationary phases were also evaluated (Cortex C18 and BEH C18). A gradient program starts with 6.4% and rises to 32.6% of phase B in 3 minutes at flow rate of 0.6 mL min-1 using Cortex C18 was found to provide the best resolution for the separation of the 20 phenolic compounds. A complete validation of the method was accomplished reaching high precisions in terms of Coefficient of Variation (CV) lower than 2% for both repeatability and intermediate precision. The suitability of the method was checked by application to the analysis of pigmented and non-pigmented rice samples. The variations in the compositions and quantities of phenolics in rice samples analyzed by the developed method disclose the effect of varietal differences. Hence, this particular chromatographic method was demonstrated to be an excellent alternative for a fast and reliable determination of phenolic compounds in a wide range of rice variety.

Comparison of HPLC and UPLC Methods for the Determination of Melatonin in Rice

Conference Paper
Setyaningsih, W., Palma, M., and Barroso, C.G
DOI: 10.13140/RG.2.1.2300.1687. The 11International Students Conference ‘ModernAnalytical Chemistry’, September 22-23, 2015, Prague, Czech Republic

Abstract

This study compares the high performance liquid chromatography (HPLC) and ultrahigh performance liquid chromatography (UPLC) methods for the determination of melatonin in rice extracts. The methods were validated in terms of linearity, precision, limits of detection (LOD) and quantification (LOQ). Repeatability and intermediate precisions expressed as coefficient of variation (CV) were less than 2.5% for HPLC and 1.5% for UPLC. Linear regression models fit perfectly the standard curves with R2 > 0.99 for both methods. The LODs were 1.15 μg L-1 and 0.73 μg L-1 while LOQs were 3.84 and 2.19 μg L-1, for HPLC and UPLC respectively. High efficiency UPLC gradient is feasible and beneficial to substitute the conventional analysis of melatonin in rice grain by HPLC as permitted the analysis run time to be reduced to one six thus consumes less eluents.

ISC

Stability of 40 phenolic compounds during Ultrasound-assisted Extractions (UAE)

Conference Paper
Setyaningsih, W., Saputro, I.E. Palma, M., and Barroso
DOI: 10.13140/RG.2.1.2955.5281. International Conference on Science and Technology 2015 (ICST2015), November 11-12, 2015, Yogyakarta, Indonesia.

Abstract

Phenolics have been positively associated with many health benefits due to their prominent antioxidant activities. Sample extractions are required prior to chromatographic measurements for the determination of phenolics in matrices. Hence, the aim of this study was to assess the stability of 40 phenolic compounds of different families (benzoic acids, benzoic aldehydes, cinnamic acids, catechins, stilbens and flavonols) under conditions of ultrasound-assisted extraction (UAE). The influence on the stability affected by the working temperature between 10 and 70°C applying ultrasounds for 20 minutes was evaluated. As for precisely pinpointing the trigger for the degradation, the extractions were repeated in light protected experiments and by stirring to check their influences on the stability of phenolics apart from the effects of ultrasounds. Regardless the present of light, all studied phenolics were stable up to 50°C with the exception for kaempferol and some light sensitive compounds such as resveratrol, sinapic acid and p-coumaric acid. By light protected UAE, starting at 60°C there were significant degradations of gallic acid (12%) and rutin (16%) whilst other compounds showed degradation starting at 70 o C i.e. p-coumaric (11%), o-coumaric (12%) and resveratrol (21%).

ICST 02

Profile of Individual Phenolic Compounds in Rice (Oryza sativa) Grains during Cooking Processes

Conference Paper
Setyaningsih, W., Saputro, I.E. Palma, M., and Barroso
DOI: 10.13140/RG.2.1.2431.2401. International Conference on Science and Technology 2015 (ICST2015), November 11-12, 2015, Yogyakarta, Indonesia.

Abstract

On top of being a prominent source of calories for more than half of the world’s population, cultivated rice (Oryza sativa) provides beneficial effects for human health through several bioactive compounds, including phenolics. To earn this advantage, it is essential to assess the profile of these compounds through appropriate cooking practices. In this study, the compositions of 18 individual phenolics contained by different cultivars (pigmented and non-pigmented rice) were evaluated during cooking processes. The level of the studied compounds dramatically decreased throughout a series of cooking processes in addition to the changes on the composition of phenolic acids and their aldehydes in the grain. Substantial losses was occurred particularly due to heating processes starting at 10 minute adjacent to the alteration of the compositions of phenolic compounds. Regardless of rice varieties, the concentration of phenolics decreased severely and remained merely 35 to 55% compared to the initial level in raw grains. This finding confirms that the stability of some phenolic compounds is adversely affected by heat. Additionally, no clear differences were found for the sensitivity of individual phenolics to degradation during the cooking steps, but most of them showed similar trend of degradation.

ICST 01

Ultrasound-assisted extraction of resveratrol from functional foods: Cookies and jams

Journal Article
Guamán-Balcázar, M.C., Setyaningsih, W., Palma, M., Barroso, C.G.
Applied Acoustics, 2016, 103 (B): 207–213, DOI:10.1016/j.apacoust.2015.07.008.

Abstract

Rapid extraction techniques with ultrasonic systems have been established to extract resveratrol from two functional foods, namely cookies and jams. The ultrasound-assisted extraction (UAE) was optimized using a full factorial design with three variables: solvent composition (10–70% and 30–90% methanol in water for cookies and jams, respectively), solvent-to-solid ratio (10:1–40:1), and ultrasonic probe diameter (2 and 7 mm). The extraction kinetics (5–30 min) were studied to confirm the full recovery of resveratrol from the functional foods. The resveratrol was quantified by Ultrahigh Performance Liquid Chromatography with a Fluorescense Detector (UPLC-FD).

The solvent composition was found to have a significant effect on the recovery (< 0.005) for both of the functional foods studied. The optimal UAE conditions were 90% methanol in water, an extraction time of 10 min, an ultrasonic probe with a diameter of 7 mm, and solvent-to-solid ratios of 40:1 and 10:1, respectively, for cookies and jams.

The methods developed in this study show a high precision in terms of Coefficient of Variance (CV), with values of less than 5% for both repeatability and intermediate precision. The applicability of the methods to real samples has been evaluated and it was confirmed that the methods are suitable for the extraction of resveratrol from cookies and jams.

Applied Acoustic 2016 02

Pressurized liquid extraction of phenolic compounds from rice (Oryza sativa) grains

Journal Article
Setyaningsih, W., Saputro, I. E., Palma, M., and Barroso, C. G
Food Chemistry, 2016, 192:452–459, DOI: 10.1016/j.foodchem.2015.06.102.

Abstract

An analytical pressurized liquid extraction (PLE) process has been studied for the extraction of phenolic compounds from rice grains. A fractional factorial design (27−2) with a centre point was used to optimize PLE parameters such as solvent composition (EtOAc in MeOH), extraction temperature, pressure, flushing, static extraction time, solvent-purge and sample weight. Extraction temperature, solvent and static extraction time were found to have a significant effect on the response value. The optimized method was validated for selectivity, linearity, limits of detection and quantification, recovery and precision. The validated method was successfully applied for the analysis of a wide variety of rice grains. Seventeen phenolic compounds were detected in the sample and guaiacol, ellagic acid, vanillic acid and protocatechuic acid were identified as the most abundant compounds. Nonetheless, different species of rice show very varied compound diversity and levels of compounds in their grain compositions.

Food Chem 2016

Optimization of the ultrasound-assisted extraction of melatonin from red rice (Oryza sativa) grains through a response surface methodology

Journal Article
Setyaningsih, W., Duros, E., Palma, M., Barroso, C.G.
Applied Acoustics, 2016, 103 (B): 129–135, DOI: 10.1016/j.apacoust.2015.04.001.

ABSTRACT:

An analytical ultrasound-assisted extraction (UAE) technique has been optimized and validated for the extraction of melatonin from rice grains. A Box–Behnken design in conjunction with a response surface methodology based on six factors and three levels was used to evaluate the effects of the studied factors prior to optimizing the UAE conditions. The significant (p < 0.05) response surface models with high coefficients of determination were fitted to the experimental data. Solvent composition and extraction temperature were found to have very significant effects on the response value (p < 0.005). The optimal UAE conditions were as follows: extraction time of 10 min, ultrasound amplitude of 30%, cycle of 0.2 s−1, extraction temperature of 40 °C, 50% methanol in water as the extraction solvent at pH 3.5 and a solvent/solid ratio 2.5:1. The method validation ensured right values for linearity, LOD, LOQ, precision and recovery. Furthermore, the method was successfully applied in the analysis of a number of rice samples throughout the rice production process. Hence, it was demonstrated that this particular UAE method is an interesting tool for the determination of melatonin in rice grain samples.

Applied Acoustic 2016 01

Determination of melatonin in rice (Oryza sativa) grains by pressurized liquid extraction

Journal Article
Setyaningsih, W., Estiono, I., Barbero, F., Palma, M., Barroso, C.G.
Journal of Agricultural and Food Chemistry, 2015, 63 (4): 1107–1115, DOI: 10.1021/jf505106m

ABSTRACT:

Melatonin provides a number of benefits for human health. The study reported here concerns the optimization, validation, and application of analytical pressurized liquid extraction and high-performance liquid chromatography coupled to a fluorescence detector for the determination of melatonin in rice grains. The factors that are most likely to affect the extraction efficiency were optimized with a 2IV7–2 fractional factorial design. The optimum extraction conditions were achieved by applying 70% EtOAc in MeOH at 200 °C and 200 atm for a static time of 5 min in two cycles with 50% flushing and a 60 s purge to extract a 2.5 g rice sample. The method validation ensured excellent linearity, limit of detection, limit of quantification, precision, and recovery. Furthermore, the method was applied to various rice products composed of polished, whole grain, aromatic, black, black glutinous, red, and parboiled rice. All kinds of pigmented rice grains showed high levels of melatonin (>100 μg kg –1), and the highest levels were found in red rice.

JAFC

 

Optimisation and validation of the microwave-assisted extraction of phenolic compounds from rice grains

Journal Article
Setyaningsih, W., Saputro, I. E., Palma, M., and Barroso, C. G
Food Chemistry, 2015, 169: 141–149. DOI:10.1016/j.foodchem.2014.07.128

ABSTRACT:

A new microwave-assisted extraction (MAE) method has been investigated for the extraction of phenolic compounds from rice grains. The experimental conditions studied included temperature (125–175 °C), microwave power (500–1000 W), time (5–15 min), solvent (10–90% EtOAc in MeOH) and solvent-to-sample ratio (10:1 to 20:1). The extraction variables were optimised by the response surface methodology. Extraction temperature and solvent were found to have a highly significant effect on the response value (p < 0.0005) and the extraction time also had a significant effect (p < 0.05). The optimised MAE conditions were as follows: extraction temperature 185 °C, microwave power 1000 W, extraction time 20 min, solvent 100% MeOH, and solvent-to-sample ratio 10:1. The developed method had a high precision (in terms of CV: 5.3% for repeatability and 5.5% for intermediate precision). Finally, the new method was applied to real samples in order to investigate the presence of phenolic compounds in a wide variety of rice grains.

Food Chem 2015

 

Study of Glutinous and Non-Glutinous Rice (Oryza Sativa) Varieties on their Antioxidant Compounds

Journal Article
Setyaningsih, W., Hidayah, N., Saputro, I. E, Palma, M., Barroso, C.G.
DOI: 10.15242/IICBE.C0115, International Conference on Plant, Marine and Environmental Sciences (PMES-2015), pp. 27-31

ABSTRACT:

Cultivated rice (Oryza sativa) is a global cereal crop in Southeast Asia. It serves as staple food, thus has a major contribution to the calorie intake. Furthermore, a number of antioxidant substances have been identified in rice including phenolic compounds and melatonin. The concentration and composition of these antioxidant compounds were studied on glutinous and nonglutinous grains for both pigmented and non-pigmented rice varieties. Additionally, several medium-amylose rice grains of three Indonesian varieties (IR64, umbul-umbul and pandan wangi) were also evaluated. Finding indicates that the composition of phenolic compounds are noticeably different between glutinous and nonglutinous rice grains. The level of both melatonin and total phenolics in non-glutinous rice was higher than its glutinous variety. Hence, higher amylose content exhibits relatively higher amount of antioxidant compounds.

PMES

Melatonin Profile during Rice (Oryza Sativa) Production

Journal Article
Setyaningsih, W., Hidayah, N., Saputro, I. E, Palma, M., Barroso, C.G.
DOI: 10.12720/joaat.1.1.60-64, Journal of Advanced Agricultural Technologies, Vol. 1, No. 1, pp. 60-64, June 2014

ABSTRACT:

Rice (Oryza sativa) is the foremost cereal crop in Southeast Asia. It serves as staple food, thus has a major contribution to the calorie intake. In addition, rice contains melatonin which is beneficial for human health. It is, therefore, essential to retain this compound by appropriate rice production processes. Melatonin profile during rice production was monitored for three varieties (IR64, umbul-umbul and pandan wangi) from conventional farming and four varieties (batang lembang, pandan wangi, black and red rice) from organic farming. The effect of polishing degree on melatonin content in rice was also evaluated. Melatonin level decreased throughout rice production and then remained steady at roughly 25-40% in final product. The most influential factor was polishing which led to melatonin losses of up to 50%. The results for organically cultivated varieties were similar. However, melatonin in black rice appeared to be persistent in the matrix during rice production.

JOAAT

Methods of Analysis of Food Components and Additives

Journal Article
Setyaningsih, W., Editor: Semih Ötleş
DOI: 10.1111/jfbc.12061, Journal of Food Biochemistry, 38: 363–364

JFB

A new microwave-assisted extraction method for melatonin determination in rice grains

Journal Article
Setyaningsih, W., Palma, M., Barroso, C.G.
Journal of Cereal Science, 2012, 56 (2): 340-346, DOI: 10.1016/j.jcs.2012.02.012.

ABSTRACT:

A new microwave-assisted extraction (MAE) method has been developed for the extraction of melatonin from rice grains. The stability of melatonin under MAE conditions was studied in order to define the working range. The studied analytical conditions for the MAE were temperature (125−175 °C), microwave power (500−1000 W), time (5−15 min), solvent (10−90% EtOAc in MeOH), and ratio of solvent to sample (10:1–20:1). Extraction variables were optimized by Response Surface Methodology (RSM). Extraction temperature was found to have a highly significant effect on the response value (p < 0.0001) and the solvent and quadratic of time also had significant effects (p < 0.1). The optimized MAE conditions were as follows: extraction temperature 195 °C, microwave power 1000 W, extraction time 20 min, solvent 100% MeOH, and ratio of solvent to sample 10:1. The developed method showed high precision (in terms of CV: 4.97% for repeatability and 4.34% for intermediate precision). Finally, the new method was applied to real samples in order to investigate the presence of melatonin in a wide variety of rice grains.

JCS

Profile of Individual Phenolics in Rice (Oryza sativa): Varietal Differences, Compounds Distribution and Composition Changes in the Grains during Rice Production

Conference Paper
Setyaningsih, W., Hidayah, N., Estiono, I., Palma, M., Barroso, C.G.
The 12th Asian Congress of Nutrition (ACN2015), May 13-18, 2015, Yokohama, Japan

Abstract

Cultivated rice (Oryza sativa) is a global staple food that subsists calories for more than half of the world’s population. Emerging evidence supports that a number of health-promoting substances have been identified in rice including phenolic compounds. Hence, individual profile of these compounds is essential to determine in both raw and processed grains. Pressurised Liquid Extraction (PLE) followed by High Performance Liquid Chromatography coupled with Photo-diode Array detector (HPLC-PDA) has been utilised as reliable analytical techniques for the extraction and quantification. Multivariate analyses based on seventeen phenolic compounds clustered the ten tested rice samples according to their species, pigments and grain productions. The level of these compounds was then individually monitored during the course of rice production of Indonesian rice varieties obtained from conventional (IR-64, umbul-umbul and pandan wangi) and organic farming (batang lembang, pandan wangi, black and red pigmented rice). Phenolics content in black pigmented rice appeared to be the most persistent in the matrix during rice production. However, in general, the level of phenolics decreased throughout a series of rice production with the most influential factors were de-husking and polishing. These particular processing steps led to phenolics losses of up to 86% as well as the changes on the composition of phenolic acids and its aldehydes in the grain. Since the profile of individual phenolics has also been evaluated on different polishing degrees (70%, 80%, 90% and 100% bran removal), the distribution of these compounds in the whole grain rice was effectively mapped. These variations in the compositions and quantities of phenolics disclosed the effect of processing steps, varietal differences as well as their distribution in the grain. Hence, the results from this study could help rice producers to optimize levels of these compounds in the final rice product by selecting the right varieties and appropriate production processes.

Study of Glutinous and Non-Glutinous Rice (Oryza Sativa) Varieties on their Antioxidant Compounds

Conference Paper
Setyaningsih, W., Hidayah, N., Estiono, I., Palma, M., Barroso, C.G
International Conference on Plant, Marine and Environmental Sciences (PMES-2015). January 1-2, 2015, Kuala Lumpur, Malaysia.

Abstract

Cultivated rice (Oryza sativa) is a global cereal crop in Southeast Asia. It serves as staple food, thus has a major contribution to the calorie intake. Furthermore, a number of antioxidant substances have been identified in rice including phenolic compounds and melatonin. The concentration and composition of these antioxidant compounds were studied on glutinous and nonglutinous grains for both pigmented and non-pigmented rice varieties. Additionally, several medium-amylose rice grains of three Indonesian varieties (IR64, umbul-umbul and pandan wangi) were also evaluated. Finding indicates that the composition of phenolic compounds are noticeably different between glutinous and nonglutinous rice grains. The level of both melatonin and total phenolics in non-glutinous rice was higher than its glutinous variety. Hence, higher amylose content exhibits relatively higher amount of antioxidant compounds

Fast Determination of Anthocyanins from Corn (Zea mays) Grains by Ultrasound-assisted Extraction

Conference Paper
Setyaningsih, W., Guamán Balcázar, M.C., Duros, E., Palma, M., and Barroso, C. G
Seminario Internacional Iberoamericanos: Propiedades Nutricionales y Funcionales. November 19-20, 2014, Loja, Ecuador

Abstract

The extraction by means of sonication method, namely ultrasound-assisted extraction (UAE), is a very interesting technique to extract natural compounds from food matrices due to the cavitation effect, which enhances mass transport by disrupting the plant cell walls. Consequently, various factors can affect the extraction process and these have to be optimized in order to extract quantitatively the analytes of interest. In this research, the chemometric approach based on the advantages of the Central Composite Design (CCD) have been successfully applied in the optimization of anthocyanins from corn (Zea mays) grains using sonication.

In conjunction with Response Surface Methodology (RSM), CCD was applied based on six factors and three levels to evaluate the significances of studied factors prior to the optimization of extraction conditions. The effects of solvent composition (25−75% methanol in water), extraction temperature (10−50oC), solvent-solid ratio (2.5:1−5:1), solvent pH (2−7), amplitude (30−70%) and cycle (0.2−0.7 s-1) on the extraction yields were taken into account. Subsequently, extraction kinetic (5−30 min) was studied to confirm the full recovery of phenolic compounds extracted from the matrix. The levels of anthocyanins in the UAE extracts were quantified by High Performance Liquid Chromatography using a Dioda Array Detector (HPLC-DAD) in less than 10 min.

The optimized condition by RSM was then assed for a kinetic study to evaluate the extraction rate. The optimal UAE condition was as follows: extraction time 5 min, ultrasound amplitude 66%, cycle 0.2 s-1, extraction temperature 10oC, 75:25 methanol:water as the extraction solvent at pH 7 and solvent-solid ratio 2.5:1. Complete validation of the method was performed. High recovery and precision were found for the full analytical method. The suitability of the method was checked by application to corn grain cultivars and demonstrated this extraction method to be suitable for the extraction of anthocyanins from corn grain samples.

Rapid ultrasound-assisted extraction (UAE) of melatonin from corn (Zea mays) grains

Conference Paper
Setyaningsih, W., Guamán Balcázar, M.C., Duros, E., Palma, M., and Barroso, C. G
V Congreso Ecuatoriano de Ingeniera en Alimentos y XIV Jornadas de Ciencia y Tecnología en Alimentos. November 25-28, 2014, Quito, Ecuador

Abstract

Corn (Zea mays) is the third most important crop worldwide and might serves as a staple food, especially in the developing countries. Moreover, corn is essential because it contains naturally occurring melatonin [N-acetyl-3-(2-aminoethyl)-5-methoxyindole]. It has a number of important activities that include neurohormone and chronobiotic actions in biological systems. There are good evidences that melatonin provides effective treatments for cardiovascular diseases, sleep disorders and headache. Reliable determination of melatonin in regular food in human diet would help to evaluate its effect. Hence, a suitable extraction method is needed to determine melatonin level in corn grains.

In this research, a new ultrasound-assisted extraction (UAE) of melatonin has been optimized and validated. Central Composite Design (CCD) in conjunction with Response Surface Methodology (RSM) was applied based on six factors and three levels to evaluate the significances of studied factors prior to optimize the UAE conditions. The effects of solvent composition (25−75% methanol in water), extraction temperature (10−50oC), solvent-solid ratio (2.5:1−5:1), solvent pH (2−7), amplitude (30−70%) and cycle (0.2−0.7 s-1) on the extraction yields were taken into account. Subsequently, extraction kinetic (5−30 min) was studied to confirm the full recovery of phenolic compounds extracted from the matrix. The levels of melatonin in the UAE extracts were quantified by Ultrahigh Performance Liquid Chromatography using a fluoresce detection (UPLC-FD) in less than 4 min.

The significant (p<0.05) response surface models with high coefficients of determination values of 0.7096 were fitted for the experimental data. Solvent composition, solvent-sample ratio and quadratic effect of temperature were found to have a greatly significant effect on the response value (p<0.05). The optimal UAE condition was as follows: extraction time 5 min, ultrasound amplitude 45%, cycle 0.7 s-1, extraction temperature 50oC, 70:30 methanol:water as the extraction solvent at pH 6 and solvent-solid ratio 2.5:1. Complete validation of the method was performed. High recovery and high precision were found for the full analytical method.

The validated method was successfully applied to the analysis of phenolics content in four corn cultivars including black, yellow, red and white. Therefore, this particularly rapid ultrasound extraction method was demonstrated to be suitable for the extraction of melatonin from corn grain samples.

Determination of Melatonin in Rice (Oryza sativa) grains: Analytical tools for Developing Melatonin-Rich Rice Products

Conference Paper
Setyaningsih, W., Palma, M., and Barroso, C. G
III Congreso Científico de Investigadores en Formación en Agroalimentación. November 18-19, 2014, Cordoba, Spain

Abstract

As a grain of life, cultivated rice (Oryza sativa) serves as the foremost food for more than half of the world’s population. Moreover, rice is essential because it contains naturally occurring melatonin [N-acetyl-3-(2-aminoethyl)-5-methoxyindole]. Recent research has led to an increase in the production of melatonin in transgenic rice seeds and melatonin-rich rice plants have been successfully generated. Functional rice products have also become more widely appreciated in the current market.

Research into melatonin was started almost a century ago and this has continued to be an active field to the present day, with many results revealing the utility of this compound. Melatonin diminishes neuro-degenerative diseases, such as Parkinson’s and Alzheimer’s diseases, and it also acts as an anticancer agent. Melatonin effects are more interesting if the compound is taken regularly at a specific time. Therefore, it should be better if people can consume melatonin-rich foods regularly instead of taking it as a drug. Rice grains, potential source of melatonin, normally contains low concentration of this compound. Hence, it is of great interest to extract and quantify endogenous levels of melatonin accurately by selecting the most suitable analytical technique.

Recent studies have shown that Microwave-assisted Extraction (MAE), Ultrasound-assisted Extraction (UAE) and Pressurized Liquid Extraction (PLE) are important sample preparation techniques and these approaches have been used to increase the extraction efficiency for various bioactive compounds. However, these advanced extraction methods have not been developed for the extraction of melatonin from rice grain yet. Thus, the methods still need to be optimised and validated to reach reliable extraction methods. As the outcome of the study, the final method could serve as an analytical tool to evaluate the level of melatonin through the production processes of rice from farm to fork.

This research prompted us to compare the performances of various extraction methods for the extraction of melatonin from rice grains including MAE, UAE and PLE, to the best of our knowledge, this kind of study has never been performed. Of the three studied extraction techniques, one may be chosen depending on the necessity and fitness for purpose.

The developed and validated method for the extraction and determination of melatonin in rice grains reported in this study will enable to come up with reliable data on the quantity of melatonin in wide variety of rice grains, which is currently lacking due to unreliable analytical data. The proposed extraction method also provides opportunities to develop advanced techniques in rice processing. Results obtained from this specific study recommended that the rice production processes should be carefully controlled in order to attain a high level of melatonin in rice products by efficient drying and the optimum degree of milling. Additionally, bearing in mind the importance of rice science and technology, it is envisaged that this work will significantly support further studies to improve rice production techniques in the coming decades.

As other products of ready to eat food, little is known about the melatonin concentration in cooked rice. Yet, several differences on its level can be expected as compared to other rice-derived products, notably in relation to the treatment during food processing. It is, therefore, the proposed analytical method was also useful to asses the level of melatonin through a series of treatments. And finally, it is noteworthy that the recommended melatonin-rich rice products, based on the result applying the analytical method, could be chosen instead of the regular ones by the population interested in the properties associated to melatonin, without dramatic changes in their regular diets.

Microwave-assisted Extraction of Resveratrol from Functional Foods: Cookies and Jams

Conference Paper
Guamán Balcázar, M.C., Setyaningsih, W., Palma, M., and Barroso, C. G
Food Analysis Congress: Safety, Quality, Novel Technologies. October 29-30, 2014, Barcelona, Spain

Abstract

Rapid extraction techniques with ultrasonic systems have been established to extract resveratrol from two functional foods, namely cookies and jams. The ultrasound-assisted extraction (UAE) was optimized using a full factorial design with three variables: solvent composition (10–70% and 30–90% methanol in water for cookies and jams, respectively), solvent-to-solid ratio (10:1–40:1), and ultrasonic probe diameter (2 and 7 mm). The extraction kinetics (5–30 min) were studied to confirm the full recovery of resveratrol from the functional foods. The resveratrol was quantified by Ultrahigh Performance Liquid Chromatography with a Fluorescense Detector (UPLC-FD).

The solvent composition was found to have a significant effect on the recovery (p < 0.005) for both of the functional foods studied. The optimal UAE conditions were 90% methanol in water, an extraction time of 10 min, an ultrasonic probe with a diameter of 7 mm, and solvent-to-solid ratios of 40:1 and 10:1, respectively, for cookies and jams.

The methods developed in this study show a high precision in terms of Coefficient of Variance (CV), with values of less than 5% for both repeatability and intermediate precision. The applicability of the methods to real samples has been evaluated and it was confirmed that the methods are suitable for the extraction of resveratrol from cookies and jams.

Fast Determination of Tryptophan and Its Derivatives by UPLC-PDA-FD

Conference Paper
Saputro, I.E., Setyaningsih, W., Palma, M., and Barroso, C. G
Food Analysis Congress: Safety, Quality, Novel Technologies. October 29-30, 2014, Barcelona, Spain.

Abstract

Tryptophan is an essential amino acid that is necessary for the biosynthesis of neurotransmitters including serotonin and melatonin via two biochemical pathways. However, further research into the role of tryptophan and its derivatives in biological systems has been limited as these compounds are mostly diverse in terms of their chemical properties resulting in the need of more than one analytical technique to determine all the derived products. To the best of recent knowledge, this is the first study that demonstrates the simultaneous identification and quantification of tryptophan and its derivatives employing the useful of ultra-high performance liquid chromatography coupled with photodiode-array and fluorescence detection (UPLC-PDA-FD). Seven compounds of tryptophan and its derivatives were effectively separated within 4 min of chromatographic analysis run time. The method was validated resulting an adequate sensitivity in the range of 0.5–20 μg L-1 (R2=0.991-0.999) with high repeatability (CV <3.9% in average) achieving very low limit of detection and quantification (0.67-2.32 μg L-1 and 2.23-7.73 μg L-1 respectively). The suitability of the method was assessed by application to the extract of ten different rice products that confirmed this rapid method to be suitable for fast determination of tryptophan and related compounds.

Optimization of ultrasound-assisted extraction (UAE) of phenolic compounds from corn (Zea mays) grains using response surface methodology

Conference Paper
Setyaningsih, W., Duros, E., Carrera, C., Palma, M., and Barroso, C. G
Food Analysis Congress: Safety, Quality, Novel Technologies. October 29-30, 2014, Barcelona, Spain

Abstract

A new Ultrasound-assisted Extraction (UAE) of phenolic compounds has been optimized and validated. Central Composite Design (CCD) in conjunction with Response Surface Methodology (RSM) was applied based on six factors and three levels to evaluate the significances of studied factors prior to optimize the UAE conditions. The effects of solvent composition (25−75% methanol in water), extraction temperature (10−50oC), solvent-solid ratio (2.5:1−5:1), solvent pH (2−7), amplitude (30−70%) and cycle (0.2−0.7 s-1) on the extraction yields were taken into account. Subsequently, extraction kinetic (5−30 min) was studied to confirm the full recovery of phenolic compounds extracted from the matrix. The levels of phenolics in the UAE extracts were quantified by Ultrahigh Performance Liquid Chromatography using a Photo-diode Array detection (UPLC-PDA) in less than 4 min of analysis run time. A complete validation of the method was performed presenting recoveries between 72 to 111% with high precisions (in average, CV was lower than 5% for both repeatability and intermediate precision). The validated method was successfully applied to the analysis of phenolics content in four corn cultivars including black, yellow, red and white. The proposed method was found to be reliable and is suited for the rapid extraction of phenolics from corn grain.

 

Melatonin Profile during Rice (Oryza sativa) Cooking: Ultimate Step to Develop a Ready to Eat Melatonin-rich Rice Food

Conference Paper
Setyaningsih, W., Palma, M., and Barroso, C. G
Food Analysis Congress: Safety, Quality, Novel Technologies. October 29-30, 2014, Barcelona, Spain

Abstract

Rice (Oryza sativa) is a pivotal staple food providing important contribution to the calorie intake for a large number of world population. Furthermore, rice contains melatonin that is favourable for human health. However, the nutritional properties of rice including melatonin differ in specific classified rice varieties. Additionally, changes in the chemical compositions of rice can also occur during the cooking processes. Hence, the aim of the study described here was to screen a wide range of rice varieties in order to develop melatonin-rich rice products. Afterwards, to evaluate the profile of melatonin during the cooking processes including soaking, boiling and warming. The levels of melatonin on the tested rice samples were decreased up to 80% in average during the course of rice cooking. However, the final levels of cooked rice from melatonin-rich (whole grain) rice were remained high, especially pigmented rice (138 µg Kg-1 in red rice and 95 µg Kg-1 in black rice) which were significantly higher than the regular (6 µg Kg-1 in white polished grain) rice. Hence, these rice varieties could be chosen instead of the regular ones by the population interested in the properties associated to melatonin, without dramatic changes in their regular diets.

Optimization of ultrasound-assisted extraction of melatonin from red rice (Oryza sativa) grain using Response surface methodology

Conference Paper
Setyaningsih, W., Duros, E., Palma, M., and Barroso, C. G
The first International conference on ultrasonic-based applications: from analysis to synthesis (ULTRASONICS-2014). September 15-17, 2014. Costa da Caparica, Portugal.

Abstract

Melatonin (N-acetyl-5-methoxytryptamine) is an indoleamine that is synthesized from the essential amino acid L-tryptophan. It has a number of important activities that include neurohormone and chronobiotic actions in biological systems. There is good evidences that melatonin provides effective treatments for cardiovascular diseases, sleep disorders and headache. Melatonin is a naturally occurring compound that is found in humans, animals, plants and microbes. The presence of melatonin in plants has been identified in a range of species including rice (Oryza sativa). Hence, a suitable extraction method is needed to determine melatonin levels in both rice and rice derived foods.

In this research, a new ultrasound-assisted extraction (UAE) of melatonin has been optimized and validated. Box-Behnken design in conjunction with response surface methodology was used based on six factors and three levels to evaluate the significances of studied factors prior to optimize the UAE condition. The effects of solvent composition (0−50% methanol in water), extraction temperature (10−70oC), solvent-solid ratio (2.5:1−5:1), solvent pH (3−7), amplitude (30−70 %) and cycle (0.2−0.7 s-1) on the extraction yields were taken into account. Subsequently, extraction kinetic (5−30 min) was studied to confirm high recovery of melatonin extracted from the matrix. The levels of melatonin in the UAE extracts were quantified by Ultrahigh Performance Liquid Chromatography using a fluoresce detection (UPLC-FD) in less than 4 min.

The significant (p<0.05) response surface models with high coefficients of determination values of 0.7245 were fitted for the experimental data. Solvent composition and extraction temperature were found to have a greatly significant effect on the response value (p<0.005). The optimal UAE condition was as follows: extraction time 10 min, ultrasound amplitude 30%, cycle 0.2 s-1, extraction temperature 40oC, 50:50 methanol:water as the extraction solvent at pH 3.5 and solvent-solid ratio 2.5:1. Complete validation of the method was performed. The developed method presented recoveries between 90% and 109% for the target analytes and showed high precision in terms of CV lower than 2.0% for intermediate precision.

 The suitability of the method was checked by application to semi-finished and finished rice products thus could assess the melatonin profile throughout the rice production. Melatonin was determined in two different rice varieties taken at different production processes i.e. drying (dried paddy), hulling (whole grain rice), and polishing (polished rice). Additionally, the effect of cultivation system on the melatonin content in rice was also able to be evaluated by applying the developed method to different rice varieties produced by conventional and organic farming of black and red rice. Hence, this particular ultrasound extraction method was demonstrated to be an excellent alternative for the extraction of melatonin from rice grain samples.

High performance allelopathy of submerged macrophytes possibly based on polyphenol complementarity

Conference Paper
Rojo, C., Rodrigo, M.A.,Carramiñana, M., Setyaningsih, W., Palma, M.
XVII Congreso de la Asociación Ibérica de Limnología. July 6-11, 2014. Santander, Spain

Abstract

Submerged macrophytes play a significant role in stabilising clear water conditions in coastal wetlands, partly due to their interference with microalgal growth via allelopathy. One of our recent studies tested allelopathic effects of several macrophyte species on microalgae. This study was carried out with the charophytes Chara hispida, C. baltica, C. vulgaris and Nitella hyalina, and with the angiosperm Myriophyllum spicatum. A higher inhibitory effect on natural microalgal assemblages was measured when charophyte exudates were present, particularly in the case of C. hispida. When mixed exudates from several populations were used, the inhibitory effects on microalgal growth were even stronger. Thus, it is likely that in aquatic communities macrophytes cause synergistic allelopathic effects on other organisms. Based on our previous results, the hypothesis that arises is whether the polyphenols of different macrophyte species with allelopathic activity are complementary rather than redundant. This fact would explain that diverse macrophyte assemblages exert a greater negative effect on the microalgae growth. To test this hypothesis, tissues of the several macrophyte species cultivated in the laboratory in monocultures were analysed for polyphenols. The analytical procedure consisted of extractions (Solid Phase Extraction Strata X Cartridge), separation (Ultra‐high Performance Liquid Chromatography (UPLC)) and setting ranges and calibration curves of compounds. The polyphenols found in the target macrophytes were: Gallic Acid, Protocatechuic Acid, Protocatechuic Aldehyde, Catechin, pOH-Benzaldehyde, Epicatechin, pCoumaric Acid, Quercetin, Epigallocatechin Gallate, pOHBenzoic Acid, Kaempferol 3-glucoside, Quercetin 3-Glucoside, Quercetin 3-Rhamnoside, Ethyl Caffeate. The number of different polyphenols in macrophytes varied from six in the angiosperm to one in C. baltica. M. spicatum’s polyphenols were mainly of the flavonoid type. Benzenoic polyphenols were the most common in charophytes. A cluster analysis (Gower index and paired group method) of the macrophyte species based on the tissue polyphenol content (quantified as mg compound/Kg DW of plant) was performed. M. spicatum formed a monospecific group separated from charophytes, and Chara species grouped together separately from N. hyalina. Thus, a complementary allelopathic effect might be likely to occur in aquatic systems dominated by dense and diverse stands of submerged vegetation.

Melatonin Profile during Rice (Oryza Sativa) Production

Conference Paper
Setyaningsih, W., Hidayah, N., Estiono, I., Palma, M., Barroso, C.G.
The 4 th International Conference on Asia Agriculture and Animal (ICAAA 2014). June 9-11, 2014. Bangkok, Thailand.

Abstract

Rice (Oryza sativa) is the foremost cereal crop in Southeast Asia. It serves as staple food, thus has a major contribution to the calorie intake. In addition, rice contains melatonin which is beneficial for human health. It is, therefore, essential to retain this compound by appropriate rice production processes. Melatonin profile during rice production was monitored for three varieties (IR64, umbul-umbul and pandan wangi) from conventional farming and four varieties (batang lembang, pandan wangi, black and red rice) from organic farming. The effect of polishing degree on melatonin content in rice was also evaluated. Melatonin level decreased throughout rice production and then remained steady at roughly 25-40% in final product. The most influential factor was polishing which led to melatonin losses of up to 50%. The results for organically cultivated varieties were similar. However, melatonin in black rice appeared to be persistent in the matrix during rice production.

Optimization and method validation of ultrasound-assisted Extraction (UAE) of serotonin and its precursor

Conference Paper
Setyaningsih, W., Carrera, C., Ruiz-Rodriguez, A., Palma, M., Barroso, C. G.
La XIV Reunión del Grupo Regional Andaluz de la Sociedad Española de Química Analítica (GRASEQA 2014). June 26-27, 2014. Baeza, Spain.

Abstract

Serotonin (5-hydroxytryptamine), a neurotransmitter in the central nervous system (CNS), is synthesised from the essential amino acid L-tryptophan by a short metabolic pathway with oxitriptan (5-hydroxytryptophan) and tryptamine as intermediate compounds. Serotonin has an enormous influence over many brain functions i.e. performing an essential role in the modulation of anger, mood, body temperature, and appetite. However, the exogenous serotonin scarcely accesses the CNS via serotonergic pathways from the bloodstream, while its precursor, oxitriptan, grants the access. Therefore, oxitriptan is recognized as dietary supplement source and has been used in clinical treatment since decades for migraine. Oxitriptan has been produced commercially by extraction from plants.

In this research, a new ultrasound-assisted extraction (UAE) of serotonin and its precursor (tryptophan, tryptamine and oxitriptan) has been optimized and validated. Box-Behnken design in conjunction with response surface methodology was used based on six factors and three levels to evaluate the significances of studied factors prior to optimize the UAE condition. The effects of solvent composition (0−50% ethanol in water), extraction temperature (10−70 oC), solvent-solid ratio (10:1−20:1), solvent pH (3−7), amplitude (30−70 %) and cycle (0.2−0.7 s-1) on the extraction yields were taken into account. Subsequently, extraction kinetic (6−18 min) was studied to reach high recovery of serotonin and its precursors extracted from the matrix. The levels of serotonin and its precursors in the UAE extracts were quantified simultaneously by Ultrahigh Performance Liquid Chromatography coupled with fluoresce detector (UPLC-FD) within 3 min of analysis run time.

The significant (p<0.05) response surface models with high coefficients of determination values of 0.78 were fitted for the experimental data. Solvent composition was found to have a greatly significant effect on the response value (p<0.0001) and the extraction temperature also have a significant effect (p <0.05). The optimal UAE condition was as follows: extraction time 6 min, ultrasound amplitude 30%, cycle 0.7 s-1, extraction temperature 40oC, water as the extraction solvent at pH 5 and solvent-solid ratio 20:1. Complete validation of the method was performed. The developed method presented recoveries between 90% and 106% for the target analytes and showed high precision in terms of CV lower than 3.5% for intermediate precision (except serotonin with 10.5%).

The suitability of the method was checked by application to Vitis vinifera L plants grown under controlled conditions, supplied with different doses of nitrogen and collected at different times during the plants growth. Hence, this particular ultrasound extraction method was demonstrated to be an excellent alternative for the extraction of serotonin and its precursors from plant sample.

Hacia productos saludables elaborados con arroces ricos en melatonina

Conference Paper
Setyaningsih, W., García, K., Rodríguez, M. C., Palma, M., Barroso, C.G.
1er. Congreso Internacional sobre Innovación y Tendencias en Procesamiento de

Resumen

El arroz (Oryza sativa) es un cereal fundamental para más de la mitad de la población mundial. Como alimento básico es un aporte calórico en la dieta, adicionalmente contiene melatonina, compuesto beneficioso para la salud. En resultados previos, los arroces; integral, negro y rojo poseen mayor nivel de melatonina comparándolos con otras muestras analizadas, por lo que estos fueron seleccionados para iniciar un estudio de desarrollo de nuevos productos con arroces ricos en melatonina. En esta etapa, se analizó el efecto de la cocción sobre el contenido en melatonina, manteniéndose ésta en niveles aun elevados: 138 μg/kg -1 en el arroz rojo, 95 μg/kg -1 en el arroz negro y 22 μg/Kg -1 en el arroz integral; en el arroz blanco apenas alcanzó 6 μg/kg -1 . La evaluación sensorial mediante prueba de calificación hedónica de los atributos globales aspecto, olor, sabor y textura mostró que no hay diferencia en la aceptación entre las 4 variedades de arroz. Mediante encuestas se analizaron otros factores que a priori podrían influir en la aceptabilidad del producto por los consumidores, tales como, la información en el etiquetado del contenido de melatonina y su origen, los mensajes sobre su efecto beneficioso en el metabolismo, y la familiaridad con la marca.

Abstract

Rice (Oryza sativa) is the essential cereal for the livelihoods of more than half of world population. Its function as staple food, makes rice has a major contribution to the calorie intake. Furthermore, rice contains melatonin that is favourable for human health. Whole grain rice (brown, black and red rice) possesses a higher level of melatonin compared to the rest of the tested rice samples. As a result, these types of rice were then developed to produce melatonin-rich rice products. The ready to eat rice in the form of boiled products were still having a relatively high level of melatonin as follows: 138 μg/Kg -1 in red rice, 95 μg/Kg -1 in black rice and 22 μg/Kg -1 in brown rice; while merely remained at 6 μg/Kg -1 in regular boiled white rice. The sensory evaluation test using hedonic rating of global attributes appearance, odor, flavor and texture showed no difference in acceptance between 4 rice varieties. Other factors that could affect the product acceptability by consumers, such as the information on the labeling of melatonin origin and its content, messages about its benefits on metabolism, and brand familiarity were analyzed by surveys.

Determinacion de compuestos fenolicos en arroz mediante extracción assistida por microondas

Conference Paper
Barroso, C.G., Setyaningsih, W., Saputro, I.E., Palma, M.
VII Congreso Ciencia y Technologia de Los Alimentos. June 12-14, 2013. Córdoba, Spain.

Allelopathy of charophytes: does it work?

Conference Paper
Rodrigo, M. A., Segura, M., Setyaningsih, W., Palma, M., Rojo, C.
6 th International Symposium on Extant and Fossil Charophytes. Nov 22 to Dec 2, 2012. Mendoza, Argentina.

Abstract

The allelopathic potential of macrophytes is extremely significant in shallow waterbodies in semiarid areas, such as the Mediterranean, where the shortage of them makes wetlands, lagoon or coastal; ponds essential aquatic ecosystems. It is also known that allelopathic effects of a macrophyte can be species-specific and this, the resistance or vulnerability of phytoplankton to macrophyte in the field will depend on the composition of both. Our goal is to investigate the allelopathic potential of charophytes that have not been studied previously in this sense (C. hispida, C. baltica, C. vulgaris, and N. hyalina) on microalga assemblaga in order to determine what type of charophyte or combination of them promotes clear water conditions through alleopathy.

Formación específica para la mejora de la comunicación en inglés del profesorado de Química Analítica de la UCA

Conference Paper
Barbero, G.F., Bellido, D., Brenes, A.M., Carmona, Y, Castro, R, Cejudo, M.J, Contero, C, Crespo, J, Cubillana, L, Díaz, M, Durán, E, El Moussaoui, N, Espada, E, Galindo, M.D, García, Granado, M.D, Guillén, D.A., Hidalgo, J.C.,Hidalgo, J.L., López M.M., López-López, J.A., Mánuel, M.V., Mendiguchía, Milla, M., Naranjo, I., Natera, R., Palacios, J.M., Pinto, J.J., Rodríguez, M.C., Ruíz, M.J., Sánchez, M., Schwarz, M., Setyaningsih, W.
Innovación Docente en Química (InDoQuím 2012), July 12-13, 2012, Barcelona, Spain

Resumen

La piedra angular del Espacio Europeo de Educación Superior (EEES) descansa en la armonización de los distintos sistemas educativos de la Unión Europea y la dotación de una forma eficaz de intercambio entre todos los estudiantes, así como la concesión de una dimensión y de una agilidad sin precedentes al proceso de cambio emprendido por las Universidades europeas.

Con la reciente entrada en vigor de los nuevos planes de estudios correspondientes a los diferentes Títulos de Grado en la Universidad de Cádiz (UCA), nuestra inserción en el EEES es ya un hecho consolidado. Dicha inserción va acompañada intrínsecamente de un proceso de internacionalización de la universidad: los crecientes convenios de intercambio de estudiantes amplían el número de alumnos que recibimos, aumentando la participación en programas de postgrado internacionales, y de ahí se deriva la necesidad de asegurar también la formación de nuestros alumnos en una competencia transversal básica y común a todos los nuevos Grados como lo es la comunicación en una segunda lengua. En el área científico-tecnológica, la lengua de referencia es el inglés.

No obstante, el proceso de internacionalización de nuestras universidades no afecta única y exclusivamente a los alumnos, sino también, al personal docente. El incremento de estudiantes Erasmus extranjeros en las aulas y en los laboratorios, la docencia en estudios de Grado y Posgrado a nivel internacional, los nuevos programas de bilingüismo en muchas Facultades y Escuelas, etc., obliga al profesorado a tomar conciencia de la necesidad de saber expresarse de forma fluida, tanto oral como escrita, desde el punto de vista técnico y en un lenguaje específico, en un segundo idioma, fundamentalmente, el inglés. La mejora del nivel competencial en dichas habilidades lingüísticas, además, le son útiles a nivel de investigación.

Es por ello que, desde hace cuatro años, llevamos desarrollando en el Departamento de Química Analítica de la UCA varios programas de formación del profesorado con vistas a perfeccionar su nivel competencial en comunicación en lengua inglesa, en el que nos hallábamos algo deficitarios de acuerdo a los autoinformes de competencias realizados. La financiación necesaria para la ejecución de estos programas de formación ha sido concedida a través de los Planes de Desarrollo del Personal Docente e Investigador (PDI) de la UCA y las Convocatorias de Actuaciones Avaladas para la Mejora Docente, Formación del Profesorado y Difusión de Resultados, del Vicerrectorado de Docencia y Formación, a través de su Unidad de Innovación Docente.

An optimization technique of pressurized liquid extraction (PLE) of melatonin from red rice (Oryza sativa) grains

Conference Paper
Setyaningsih, W., Barbero, Gerardo., Palma, M., Barroso, C.G.
La XIII Reunión del Grupo Regional Andaluz de la Sociedad Española de Química Analítica (GRASEQA 2012). June 7-8, 2012. Malaga, Spain.

Abstract

An analytical pressurized liquid extraction (PLE) process has been studied for the extraction of phenolic compounds from rice grains. A fractional factorial design (27−2) with a centre point was used to optimize PLE parameters such as solvent composition (EtOAc in MeOH), extraction temperature, pressure, flushing, static extraction time, solvent-purge and sample weight. Extraction temperature, solvent and static extraction time were found to have a significant effect on the response value. The optimized method was validated for selectivity, linearity, limits of detection and quantification, recovery and precision. The validated method was successfully applied for the analysis of a wide variety of rice grains. Seventeen phenolic compounds were detected in the sample and guaiacol, ellagic acid, vanillic acid and protocatechuic acid were identified as the most abundant compounds. Nonetheless, different species of rice show very varied compound diversity and levels of compounds in their grain compositions.