Effects of binary solvent extraction system,extraction time and extraction temperature on phenolic antioxidants and antioxidant capacity from mengkudu (Morinda citrifolia)

An investigation into the effects of ethanol concentration (0–100%, v/v), extraction time (20–120 min) and extraction temperature (25–65 °C) on the extraction of phenolic antioxidants from mengkudu (Morinda citrifolia) was performed using a single-factor experiment. Total phenolic content (TPC) and total flavonoid content (TFC) assays were used for determination of phenolic compounds. Antioxidant capacity was evaluated by measuring the scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and 2,2′-diphenyl-1-picrylhydrazyl (DPPH) radicals. Experimental results showed that extraction conditions had significant effect on extraction of phenolic compounds and antioxidant capacities. The optimised conditions were 40% ethanol for 80 min at 65 °C, with values of 919.95 mg GAE/100 g DW for TPC, 472.73 mg CE/100 g DW for TFC, 791.71 μmol TEAC/100 g DW for ABTS and 1928.5 μmol TEAC/100 g DW for DPPH. TPC was significantly correlated with DPPH under the effects of ethanol concentration (r = 0.932) and extraction time (r = −0.938).     

Microwave-assisted extraction of phenolics with maximal antioxidant activities in tomatoes

Microwave-assisted extraction (MAE) was investigated for extraction of phenolic compounds from tomato with maximised antioxidant activities using response surface methodology (RSM) coupled with a central composite design, and in vitro antioxidant assays (FRAP and ORAC). MAE was more efficient for greater antioxidant activities and higher total phenolic contents than solvent extraction. The optimal MAE processing parameters were 96.5 °C, 2.06 min, 66.2% ethanol for FRAP, and 96.5 °C, 1.66 min, 61.1% ethanol for ORAC. The models were successfully applied to 20 tomato cultivars, whose total phenolic contents (TPC) and indexes (TPI) were 489.30–997.45 mg gallic acid equivalent/100 g dry weight (DW) and 281.34–468.52 mg/100 g DW, respectively. Eight phenolic compounds were identified. Individual phenolics were 6.10–42.73 mg/100 g DW. The FRAP, but not the ORAC value showed good correlation with the TPC or TPI. The methodologies developed and the knowledge acquired in this study will provide useful information to tomato breeders and food processors.     

The optimisation of extraction of antioxidants from potato peel by pressurised liquids

Response Surface Methodology was used to optimise the solid–liquid extraction and Pressurised Liquid Extraction of polyphenols from industrially generated potato peel. Efficiency of extraction was optimised by measuring antioxidant activity, phenol content and the level of caffeic acid. Conditions for optimal antioxidant activity as measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay were 75% ethanol, 80 °C and 22 min with solid–liquid extraction, resulting in an optimum activity of 352 mg Trolox Equivalents/100 g DW potato peel. In comparison, the use of Pressurised Liquid Extraction resulted in an optimum activity of 339 mg Trolox Equivalents/100 g DW potato peel at 70% ethanol and 125 °C. Therefore the use of Pressurised Liquid Extraction did not enhance extraction in comparison to solid–liquid extracts, but using aqueous ethanol as extraction solvent recovered a higher level of polyphenols than when using 100% methanol.     

Optimization of tyrosinase inhibition activity of ultrasonic-extracted polysaccharides from longan fruit pericarp

Various ultrasonic conditions were employed to prepare polysaccharides from longan fruit pericarp (PLFP) and the Lineweaver–Burk equation was then used to determine the effect of PLFP on inhibition of tyrosinase activity. This result showed that PLFP acted as a non-competitive inhibitor of tyrosinase. The highest slope was observed for ultrasonic extraction, followed by the hot-water extraction, suggesting that the ultrasonic treatment of PLFP increased the inhibition of tyrosinase activity. Furthermore, a multilayer feed-forward neural network trained with an error back-propagation algorithm was used to evaluate the effects of ultrasonic power, time and temperature on the slope value. The trained network gave a regression coefficient (R²) of 0.98 and a mean squared error (MSE) of 0.58, implying a good agreement between the predicted value and the actual value of the slope, and confirmed a good generalization of the network. Based on the artificial neural network-genetic algorithm, the optimal ultrasonic extraction conditions to obtain the highest slope value (154.1) were determined to be 120 W, 12 min and 57 °C. Application of response surface plots showed the slope value as a function of every two factors under various ultrasonic extraction conditions, which can be observed directly. Therefore, the artificial neural network provided a model with high performance and indicated the non-linear nature of the relation between ultrasonic conditions and slope value.     

The optimisation of solid–liquid extraction of antioxidants from apple pomace by response surface methodology

Response surface methodology using two food grade solvents, acetone and ethanol, was used to optimise antioxidant extraction from industrially generated apple pomace. Efficiency of extraction was optimised by measuring antioxidant activity, phenol content and three individual polyphenol groups. Conditions for optimal antioxidant activity as measured by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay were 56% ethanol, 80 °C and 31 min. Using these conditions an antioxidant value of 444 mg Trolox/100 g DW was obtained. For acetone extraction the optimal conditions were 65% acetone, 25 °C and 60 min, resulting in an antioxidant value of 436 mg Trolox/100 g DW. Both ethanol and acetone would be suitable to replace methanol for a food grade and more environmental friendly solid–liquid extraction of antioxidants.     

Effects of phosphorus supply on the quality of green tea

Self-rooted, 10-month-old, uniform tea [Camellia sinensis (L.) O. Kuntze cv. Huangguanyin] plants were supplied with 0, 40, 80, 160, 400 or 1000 μM phosphorus (P) for 17 weeks to determine how P-deficiency affects the quality of green tea. Leaf P concentration increased with increasing P supply. Whole plant dry weight (DW) increased as P supply increased from 0 to 160 μM, then remained little changed with further increasing P supply. The P-deficient green tea displayed decreased concentrations of water extract, total polyphenols, flavonoids, total free amino acids, theanine (Thea) and asparagic acid (Asp) + glutamic acid (Glu), increased concentrations of water soluble sugars, valine (Val), γ-aminobutyric acid (GABA), proline (Pro) and cysteine (Cys), and ratio of total polyphenols to total free amino acids, but unchanged concentrations of total catechins and epigallocatechin gallate (EGCG). In conclusion, the sensory and biochemical qualities of green tea were lowered by P-deficiency.     

Extraction of pepsin-soluble collagen from grass carp (Ctenopharyngodon idella) skin using an artificial neural network

A multilayer feed-forward neural network trained with an error back-propagation algorithm was used to evaluate the effects of pepsin amount, reaction time and pH on the yield of pepsin-soluble collagen. A positive correlation was observed between the yield and the amount of pepsin and also the reaction time. The yield increased with an increase of pH to nearly 3, thereafter yield decreased. The trained network gave a regression coefficient (r²) of 0.97 and a mean squared error (MSE) of 0.21, which implied a good generalisation of the network. Based on the genetic algorithm, the optimal extraction conditions to obtain the highest yield were determined to be pH 3.4, 53.3 unit/mg of pepsin and 35.2 h. The predicted yield value was 30.3%. As the estimated optimal extraction conditions were used in the actual preparation of the pepsin-soluble collagen, the yield was measured experimentally to be 29.3 ± 0.8%, which was not significantly different (p > 0.05) from the predicted value. The response surface plots showed the yield of pepsin-soluble collagen as a function of two factors under various extraction conditions.     

Optimization of microwave-assisted extraction of phenolics from potato and its downstream waste using orthogonal array design

While other extraction methods have been tempted, a microwave-assisted extraction (MAE) method coupled with the orthogonal array design was investigated for efficient extraction of the phenolic compounds in potato downstream wastes. Four parameters were examined for the MAE of the total phenolic content (TPC) and optimized at 60% ethanol, 80 °C, 2 min, solid-to-solvent ratio 1:40 (g/ml). The MAE was proven more efficient than the conventional solvent extraction by refluxing. The optimized model showed that the downstream wastes, both the supernatant and the residue contained high TPC, particularly the former (11.0 ± 0.26 mg GAE/g DW). The antioxidant activities (DPPH and FRAP) closely correlated with the TPC of the samples (r = 0.92–0.97). Chlorogenic acid and caffeic acid were found to be the predominant phenolic acids. The extracts of the downstream wastes from potato processing can be a promising candidate for functional foods and nutraceutical ingredients.     

Green tea: A promising anticancer agent for renal cell carcinoma

Renal cell carcinoma (RCC) is one of the most lethal amongst the urologic malignancies, comprising three percent of all human neoplasms, and its incidence appears to be rising. RCC is refractory to both chemotherapy and radiotherapy. Therefore, the discovery of new strategies for therapeutic intervention remains a priority. Green tea (Camellia sinensis) and tea polyphenols have been proposed to exert protective effects against several types of cancer, based on preclinical and clinical trial data; however, the anticarcinogenic activity of green tea towards RCC is unknown. In this study, a targeted metabolite analysis on a green tea leaves methanolic extract was performed by HPLC/DAD and the antiproliferative activity of the extract was assayed using human renal cancer cell lines A-498 and 769-P. The total phenolic content was very high (31.8% of methanolic extract), and the main compounds were flavan-3-ols (94.3% of the total phenolic content), and especially (−)-epigallocatechin-3-gallate (35.9% of the total phenolic content). In addition, two methylxanthines – theophylline and caffeine – were also present in the extract, caffeine being the most abundant. Green tea extract strongly inhibited the growth of both RCC cell lines in a concentration-dependent manner, with IC₅₀ values of 54 ± 10 and 129 ± 28 μg/ml for A-498 and 769-P cells, respectively. This is the first report showing that green tea is likely to be an effective anticancer agent for renal cell carcinoma.     

Melatonin contents in mulberry (Morus spp.) leaves: Effects of sample preparation,cultivar, leaf age and tea processing

The effects of sample preparation, cultivar, leaf age and tea processing on melatonin content of mulberry (Morus spp.) leaves were investigated. Sample preparation using ultrasonic technique in combination with solid phase extraction resulted in high recovery rate (76%), when compared to homogenisation in combination with liquid–liquid extraction procedure (12% recovery rate). The melatonin contents in mulberry leaves harvested from three major cultivars (Buriram 60, Sakonnakhon and Khunphai) grown in Thailand were identified and quantified using high-performance liquid chromatography combined with fluorescence detector. The average melatonin content of the mulberry leaves cv. Buriram 60 (279.6 ng/g dry weight (DW)) was higher than those of cv. Sakonnakhon (100.5 ng/g DW) and cv. Khunphai (40.7 ng/g DW). The melatonin contents of all cultivars tested were highest in the tip of the leaves, followed by that in the young leaves, whereas the lowest was found in the old leaves. The melatonin contents of the two types of tea produced from mulberry leaves cv. Buriram 60 were also determined. Heat treatment during tea processing decreased the melatonin content in mulberry leaves cv. Buriram 60 by approximately 87%, when compared to that of the fresh leaves. However, there were no significant differences between the melatonin contents of the mulberry leaf tea produced with blanching (mulberry green tea) and those produced without blanching (mulberry black tea).     

Effects of cosolvents on the decaffeination of green tea by supercritical carbon dioxide

Due to the adverse effects of the caffeine in a variety of plant products, many methods have been explored for decaffeination, in efforts to remove or reduce the caffeine contained in plant materials. In this study, in order to remove caffeine from green tea (Camellia sinensis) leaves, we have employed supercritical carbon dioxide (SC–CO₂), which is known to be an ideal solvent, coupled with a cosolvent, such as ethanol or water. By varying the extraction conditions, changes not only in the amount of caffeine, but also in the quantities of the principal bioactive components of green tea, including catechins, such as epigallocatechin gallate (EGCG), epigallocatechin (EGC), epicatechin gallate (ECG) and epicatechin (EC), were determined. The extraction conditions, including temperature, pressure and the cosolvent used, were determined to affect the efficacy of caffeine and catechin extraction. In particular, the type and concentration of a cosolvent used constituted critical factors for the caffeine removal, combined with minimal loss of catechins, especially EGCG. When the dry green tea leaves were extracted with SC–CO₂ modified with 95% (v/v) ethanol at 7.0 g per 100 g of CO₂ at 300 bar and 70 °C for 120 min, the caffeine content in the decaffeinated green tea leaves was reduced to 2.6% of the initial content. However, after the SC–CO₂ extraction, a substantial loss of EGCG, as much as 37.8% of original content, proved unavoidable.     

Extraction of active ingredients from green tea (Camellia sinensis): Extraction efficiency of major catechins and caffeine

The effect of different extraction set-ups that influence the extraction efficiency of catechins and caffeine from green tea leaves (variety Fanning Belas, China) were studied using different aqueous and pure solvents (acetone, ethanol, methanol, acetonitrile, water), different temperatures (60, 80, 95 and 100 °C) and times (5–240 min). Raw extracts were analysed for contents of major catechins (EC, EGC, ECG, EGCG), caffeine, proanthocyanidins and flavonols (myricetin, caempherol, quercetin). Starting material was found to contain 191 g major catechins/kg material, 36 g caffeine/kg material and 5.2 g flavonols/kg material on a dry mass basis. The content of major catechins in green tea extracts varied from approximately 280–580 g/kg dry extract, with extraction efficiencies of major catechins varying from 61% to almost 100%. Content of caffeine in extract was in the range of 75 g/kg, where its extraction efficiency varied from 62% to 76%. Average extraction yield was 30% with exceptions when using pure acetone and acetonitrile, where extraction yield was about 3%. Contents of flavonols and proanthocyanidins were in the ranges 6–20 and 12–19 g/kg, respectively. Different extraction procedures with water were also investigated and optimal conditions determined: maximum achieved extraction efficiency of catechins with water was obtained at 80 °C after 20 min (97%) and at 95 °C after 10 min of extraction (90%). Degradation of catechins was observed at higher extraction temperatures and with prolonged extraction times. Using a lower ratio of solvent to material, extraction efficiencies were increased by applying a multi-step extraction procedure. Optimal extraction procedure was then performed using decaffeinated green tea leaves, which were obtained by high-pressure extraction with CO₂, when 98% of caffeine was selectively isolated without significant impact on valuable catechins.     

Polyphenol and glycoalkaloid contents in potato cultivars grown in Luxembourg

The polyphenol (phenolic acids, flavanols and flavonols) and glycoalkaloid (α-chaconine and α-solanine) contents of potato tubers grown in Luxembourg were analyzed by UPLC–DAD and HPLC–MS/MS separately in peel (approx. 2 mm), outer (approx. 1 cm) and inner flesh. Polyphenol contents decreased from the peel via the outer to the inner flesh and differed among the cultivars. The cultivars Vitelotte and Luminella had the highest polyphenol contents (5202 and 572 μg/g dry weight (DW) in the outer flesh), whereas Charlotte and Bintje had the lowest contents (19.5 and 48.0 μg/g DW). Chlorogenic acid and its isomers (neo- and cryptochlorogenic acid) were the major polyphenols. Glycoalkaloid contents were highest in the peel and lowest in the inner flesh, values in the flesh were below guideline limits in all cultivars. In conclusion, potatoes contribute to the daily intake of polyphenols and their consumption, thereby, may have positive effects on health.     

The composition of polyphenols and methylxanthines in teas and herbal infusions

The popularity of tea is increasing on the global aspect because of its role as a significant source of phenolic compounds in human diet. The purpose of this study was to determine and compare the phenolic and methylxanthine composition as well as the antioxidant capacity of white, green, Oolong and black teas, and chamomile and linden infusions depending on the extraction conditions (water temperature and multiple extractions). The content of total phenols and total flavonoids in teas and herbal infusions was determined by using UV/vis spectrophotometric methods, whilst individual polyphenols (phenolic acids and flavan-3-ols) and methylxanthines were identified and quantified by using high performance liquid chromatography coupled with photodiode array detection. In order to determine the antioxidant capacity of teas the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical scavenging assays were applied. The highest content of phenolic compounds was determined in green tea, which also demonstrated the highest antioxidant capacity, whilst herbal infusions were characterised with the lowest content of phenolic compounds, as well as the lowest antioxidant capacity. The highest content of caffeine, as the most abundant methylxanthine, was determined in black tea. Extraction at 100 °C is the most effective to extract the highest content of polyphenols and methylxanthines in all studied teas.     

Ultrasound-assisted extraction of polyphenols (flavanone glycosides) from orange (Citrus sinensis L.) peel

The present study reports on the extraction of polyphenols especially flavanones from orange (Citrus sinensis L.) peel by using ethanol as a food grade solvent. After a preliminary study showing that the best yield of extraction was reached for a particle size of 2 cm², a response surface methodology (RSM) was launched to investigate the influence of process variables on the ultrasound-assisted extraction (UAE) followed by a central composite design (CCD) approach. The statistical analysis revealed that the optimised conditions were a temperature of 40 °C, a sonication power of 150 W and a 4:1 (v/v) ethanol:water ratio. The high total phenolic content (275.8 mg of gallic acid equivalent/100 g FW), flavanone concentrations (70.3 mg of naringin and 205.2 mg of hesperidin/100 g FW) and extraction yield (10.9 %) obtained from optimised UAE proved its efficiency when compared with the conventional method. Furthermore, the antioxidant activity determined by the DPPH and ORAC tests confirmed the suitability of UAE for the preparation of antioxidant-rich plant extracts.     

Pressurised water extraction of polyphenols from pomegranate peels

Pomegranate peels are one of the most valuable by-products of the food industry in terms of polyphenols which are conventionally extracted from plant materials by organic solvents, especially with methanol. Pressurised water extraction was investigated for the extraction of polyphenols from pomegranate peels. The most important factors affecting the extraction results were found to be particle size, temperature, and static time. The results indicated that pressurised water extraction was as effective as conventional methanol extraction for the recovery of polyphenols from pomegranate peels. Total phenolic contents of pomegranate peels obtained by pressurised water extraction at optimised conditions and conventional solid–liquid methanol extraction were determined as 264.3 and 258.2 mg/g tannic acid equivalents, respectively. Hydrolyzable tannins were the predominant polyphenols of pomegranate peels corresponding to 262.7 mg/g tannic acid equivalents. Punicalagin content of pomegranate peels by pressurised water extraction was found to be 116.6 mg/g on dry matter basis.     

Simultaneous spectrophotometric determination of caffeine and theobromine in Iranian tea by artificial neural networks and its comparison with PLS

In order to determine the amount of caffeine and theobromine, spectrophotometry was used as a simple, rapid and economical method. Because of severe overlapping between these components, artificial neural network was used. The 230–300 nm spectral window with 1 nm interval was used for data acquisition. An artificial neural network (5-5-3) with linear transfer function between input-hidden and hidden-output layers was trained and applied for prediction of concentration of these methylxanthines in four Iranian tea samples. The model was compared with PLS modeling method. HPLC technique was used as a standard method.     

Changes of phenolic acids and antioxidant activities during potherb mustard (Brassica juncea, Coss.) pickling

Phenolic acids in potherb mustard (Brassica juncea, Coss.) were determined and the effects of pickling methods on the contents of total free phenolic acids, total phenolic acids, total phenolics, and antioxidant activities were investigated. Gallic acid, protocatechuic acid, p-hydroxybenzoic acid, vanillic acid, caffeic acid, p-coumaric acid, ferulic acid, and sinapic acid were identified in the present study. The contents of total free phenolic acids, total phenolic acids and total phenolics in fresh potherb mustard were 84.8 ± 0.58 μg/g dry weight (DW), 539 ± 1.36 μg/g DW, and 7.95 ± 0.28 mg/g DW, respectively. The total free phenolic acids increased during the pickling processes, but the total phenolic acids, total phenolics, and antioxidant activities decreased. However, after 5 weeks of fermentation, all the pickling methods retained over 70% of total phenolic contents and above 65% of antioxidant capacities. The results indicated that pickling processes were relatively good methods for the preservation of phenolic acids and antioxidants for potherb mustard.     

Antioxidant properties of chlorophyll-enriched and chlorophyll-depleted polyphenolic fractions from leaves of Vernonia amygdalina and Gongronema latifolium

This study investigated the optimum polyphenol extraction conditions for two Nigerian green leafy vegetables, Vernonia amygdalina (VA) and Gongronema latifolium (GL) that are widely used for food and medicinal purposes. Seven different solvents were used to extract the total phenolic contents of the leaves; 100% acetone, 80% acetone, 50% acetone, 100% ethanol, 70% ethanol, 100% methanol and 70% methanol and their total phenolic contents were determined. The results showed that the 80% acetone was significantly (p < 0.05) the most effective for polyphenol extraction from the leaves and generally the acetone-containing solvents were more effective than alcohol-containing solvents. The 80% acetone extract was fractionated on a silicic acid-packed column to give two main fractions: acetone eluted chlorophyll-enriched (flow-through) and ethanol eluted chlorophyll-depleted (column-bound). The ethanol fractions of VA and GL were significantly better (p < 0.05) DPPH radical scavengers than the acetone fractions. In contrast, the acetone fraction was a significantly better (p < 0.05) scavenger of hydroxyl radicals when compared to the ethanol fraction. Both acetone and ethanol fractions significantly (p < 0.05) and dose-dependently inhibited linoleic acid oxidation, though duration of inhibition was less at lower concentrations (10 and 25 ??g/ml) than at the highest concentration (50 ??g/ml).     

Column-chromatographic extraction and separation of polyphenols, caffeine and theanine from green tea

A highly efficient column-chromatographic extraction (CCE) followed by sequential adsorption to extract and separate bioactive compounds from green tea was developed. Tea powder was loaded into columns with 4-fold solvents and eluted through a cyclic CCE. High-quality tea extracts with greater than 90% extraction efficiencies of polyphenols, epigallocatechin-3-gallate, caffeine, theanine and polysaccharides were obtained with 4-fold water circulated five times among different columns at 70 °C. Similar results, except for low polysaccharide extraction (35.5%), were obtained with 4-fold 30% ethanol circulated three times at room temperature. The highly concentrated water extraction was directly passed through columns of polyamide, DM130 macroporous and 732 ion exchange resins, resulting in high-purity polyphenols (99%), caffeine (98%) and theanine (98%) after simple purification of the eluates from each column. This method uses simple equipment, minimum solvents and can be used for both quantitative analysis and continuous preparation of high-quality tea extracts and bioactive compounds.