In vitro screening of phenolic compounds, potential inhibition against α-amylase and α-glucosidase of culinary herbs in Thailand

Herbs that are commonly used in Thai dishes were selected for evaluation in order to determine their phenolic compounds, antioxidant activity, and in vitro potential inhibition against α-amylase and α-glucosidase. The total phenolic content ranged from 2.89 to 75.26 mg GAE/g dw. The three aqueous herb extracts with the highest total phenolic content were yellow-berried nightshade (Solanum xanthocarpum), holy basil (Ocimum sanctum) and acacia (Acacia pennata). The antioxidant activity was expressed as percent inhibition of DPPH, ranging from a high of 85% in bitter gourd (Monordica charantia) to a low of 0% in garlic (Allium sativum) and shallot (Allium cepa). A high correlation (r = 0.70, p < 0.05) was observed between total phenolic content and antioxidant activity for the herb extracts in the Solanaceae family while, an insignificant high and negatively correlation (r = −1.00, p > 0.05) for the herb extracts in the Cucurbitaceae and Umbelliferae families was observed. All of the herbs had caffeic content, varying from 0.00 to 23.93 mg/g dw. Peppermint (Mentha canalenisa), galangal (Languas galangal) and holy basil (O. sanctum) had a significant p-coumaric acid content. The first and second obtained principal component represented 60% of the total variation. The potential inhibition against α-amylase for the herbs extracts ranged from 0% to 58%, while the highest percentage was observed in the acacia (A. pennata) extract. The potential inhibition against α-glucosidase varied from 7% to 100%. A high correlation (r = 0.68, p < 0.05) was observed between α-amylase inhibition and caffeic acid content for aqueous extracts in the Alliaceae, Cucurbitaceae, and Leguminosae families, while a high correlation (r = 0.49, p < 0.05) was observed between α-glucosidase inhibition and total phenolic content for the aqueous extracts of all herbs. Therefore, specific culinary herbs showed to have a potential use for dietary management during the early stages of hyperglycaemia.     

Enrichment of dry-cured ham with α-linolenic acid and α-tocopherol by the use of linseed oil and α-tocopheryl acetate in pig diets

The use of α-linolenic acid and α-tocopherol enriched pork on the fatty acids and the sensory characteristics of Spanish dry-cured hams have been studied. Five batches of hams were manufactured using the posterior legs of pigs fed on diets with the same ingredients except for the oil source: sunflower (C), linseed (L) or linseed and olive (1/1, w/w, LO). Two different α-tocopheryl acetate concentrations [20 (C, L and LO) or 220 (LOE and LE)mg/kg diet] were used. Biceps femoris and Semitendinosus/Semimembranosus muscles from hams with low polyunsaturated fatty acid n-6/n-3 ratio (less than 3) were obtained from animals fed on linseed and linseed/olive oil enriched diets. However, hams from animals fed on diets added with linseed and α-tocopheryl acetate (20mg/kg diet) (batch L) were rejected by consumers because of less acceptable sensory characteristics and higher TBARs. The remaining hams had satisfactory sensory and nutritional characteristics.     

Bean cultivars (Phaseolus vulgaris L.) have similar high antioxidant capacity,in vitro inhibition of α-amylase and α-glucosidase while diverse phenolic composition and concentration

Common beans are a good source of essential nutrients such as protein, fiber, vitamins, and minerals; they also contain phenolic compounds and other phytochemicals. Phenolic compounds exhibit high antioxidant capacity that promotes health benefits by reducing oxidative stress. The objective was to compare the composition and quantity of anthocyanins and other non-colored phenolic compounds in fifteen improved bean cultivars from Mexico and Brazil and their relation to antioxidant capacity and enzymes related to type-2 diabetes. Samples were analyzed for total phenolic compounds (TP), flavonoids, antioxidant capacity (AC), tannins and total anthocyanins. Type and quantity were evaluated by HPLC-ESI-MS. Delphinidin glucoside (0.9–129.0 mg/100 g dry coat), petunidin glucoside (0.7–115.0 mg/100 g dry coat) and malvidin glucoside (0.14–52.0 mg/100 g dry coat). Anthocyanidins were positively correlated when quantified by HPLC and colorimetric analysis (R = 0.99). Cultivar Negro-Otomi presented the highest concentration of anthocyanins (250 mg/100 g dry coat). Seventeen non-colored phenolic compounds were identified among cultivars; catechin, myricetin 3-O-arabinoside, epicatechin, vanillic acid, syringic acid and o-coumaric acid, presented the highest concentrations among identified phenolic compounds. The AC of all fifteen bean cultivars did not show significant differences (p > 0.05) ranging from 185.2 (FM-67) to 233.9 (FM-199) mmol TE/g coat. Compounds in the coat extracts of pinto and black cultivars were the most efficient to inhibit α-amylase and α-glucosidase. Studied cultivars differed in composition and concentration of phenolics including anthocyanins; however, there was no effect on AC as measured by oxygen radical absorbance capacity. Black beans contained delphinidin and ferulic acid, compounds commonly used as ingredients in functional foods due to their associated health benefits.     

Determination of free α-lipoic acid in foodstuffs by HPLC coupled with CEAD and ESI-MS

A simple and rapid method for determination of free α-lipoic acid in different food matrices has been developed. It consists of extraction of α-lipoic acid with 0.5% glacial acetic acid in methanol by sonication, quantitative analysis of the extract by isocratic RP-HPLC (acetonitrile/methanol/50 mM potassium dihydrogen phosphate buffer adjusted to pH 3 with phosphoric acid: 350/65/585, v:v:v) at a flow rate of 0.45 ml/min coupled with coulometric electrode array detection at potentials between +300 and +700 mV and qualitative analysis by LC–ESI-MS in the negative ion mode for confirmation. Egg, dried egg powder, mayonnaise, fine peas and potatoes were analysed and free α-lipoic acid contents ranged from 0.1 to 4.2 μg/g with recoveries between 70% and 94%. Limits of quantitation were between 0.1 and 0.3 μg/g. This newly developed method can be used to establish a database for the content of free α-lipoic acid in different foodstuffs.     

The α-amylase and α-glucosidase inhibitory effects of Irish seaweed extracts

To date, numerous studies have reported on the antidiabetic properties of various plant extracts through inhibition of carbohydrate-hydrolysing enzymes. The objective of this research was to evaluate extracts of seaweeds for α-amylase and α-glucosidase inhibitory effects. Cold water and ethanol extracts of 15 seaweeds were initially screened and from this, five brown seaweed species were chosen. The cold water and ethanol extracts of Ascophyllum nodosum had the strongest α-amylase inhibitory effect with IC₅₀ values of 53.6 and 44.7 μg/ml, respectively. Moreover, the extracts of Fucus vesiculosus Linnaeus were found to be potent inhibitors of α-glucosidase with IC₅₀ values of 0.32 and 0.49 μg/ml. The observed effects were associated with the phenolic content and antioxidant activity of the extracts, and the concentrations used were below cytotoxic levels. Overall, our findings suggest that brown seaweed extracts may limit the release of simple sugars from the gut and thereby alleviate postprandial hyperglycaemia.     

Investigation of α-glucosidase inhibitory activity of wheat bran and germ

Postprandial hyperglycaemia is a primary risk factor in the development of Type 2 diabetes. α-Glucosidase inhibitors that reduce postprandial hyperglycaemia have a key role in the treatment of Type 2 pre-diabetic states and also have the potential to reduce the progression to complications of diabetes. Epidemiological studies showed that risk for Type 2 diabetes mellitus was decreased with consumption of whole grains. The bran and germ of whole wheat are major components of whole grain consumption and are widely accepted as important ingredients in many low glycaemic index (GI) foods. In this study, the α-glucosidase inhibitory activity of wheat bran and germ was investigated. The active compounds were screened using an in vitro enzyme-inhibitory assay guided fractionation. Potent α-glucosidase inhibitory compounds from wheat germ were identified as phosphatidic acids, 1,2-dilinoleoylglycero-3-phosphate and 1-palmitoyl-2-linoleoyl-glycero-3-phosphate. The low GI property of whole grain wheat could be attributed to these α-glucosidase inhibitory phosphatidic acids, which have the potential to prevent or treat Type 2 diabetes.     

Metabolite profiling and inhibitory properties of leaf extracts of Ficus benjamina towards α-glucosidase and α-amylase

The use of antioxidant-rich medicinal plants having the potential to reduce oxidative stress and postprandial hyperglycemic pressure is one of the most promising option for the management of diabetes. This study presents information on metabolite profiling and in vitro anti-diabetic effects of leaf extracts of Ficus benjamina. The DPPH (2, 2-diphenyl-1-picrylhydrazyl radicals) assay was performed to determine the in vitro antioxidant potential of the plant extracts. The anti-diabetic effects were investigated by evaluating inhibitory properties of F. benjamina leaf extracts towards carbohydrate hydrolyzing enzymes, i.e., α-glucosidase and α-amylase, whereas ¹H NMR and UHPLC-QTOF-MS/MS analytical methods were employed for metabolite profiling of F. benjamina leaf extracts. Among 40, 60, 80, and 100% ethanolic leaf extracts of F. benjamina, 80% ethanolic extract exhibited the highest antioxidant activity based upon its DPPH radical scavenging ability (IC₅₀ value: 63.71 ± 2.66 µg/mL). The 80% ethanolic leaf extract of F. benjamina also proved to be the most efficient α-glucosidase and α-amylase inhibitor with IC₅₀ values of 9.65 ± 1.04 µg/mL and 13.08 ± 1.06 µg/mL, respectively; these values were even better than acarbose with α-glucosidase inhibition activity (IC₅₀ = 116.01 ± 3.83 µg/mL) and α-amylase inhibition activity (IC₅₀ = 152.66 ± 7.32 µg/mL). Moreover, a total of 31 metabolites were identified in F. benjamina leaf extract, which may have the potential to contribute to its antioxidant and inhibitory properties against carbohydrate hydrolyzing enzymes. The findings of this study depict F. benjamina leaf extracts as a promising α-glucosidase and α-amylase inhibitor, and therefore, can be utilized for the development of anti-diabetic functional diets/nutra-pharmaceuticals.     

Study of nonenzymic browning in α-amino acid and γ-aminobutyric acid/sugar model systems

The reactivities, in nonenzymic browning, of γ-aminobutyric acid (GABA), a non-protein amino acid with wide natural occurrence and potential health benefits as it occurs in foods, and of the α-l-amino acids arginine, glutamic acid, glutamine, leucine, lysine, and phenylalanine were investigated by heating equimolar mixtures of glucose and the cited amino acids at 110 °C at pH 6.0 for different times (0–4 h). Linear regression analysis indicated that the colour development in a GABA/glucose mixture was slower than that of a lysine/glucose mixture and comparable to that of a phenylalanine/glucose mixture. High-performance anion-exchange chromatography (HPAEC) with integrated pulsed amperometric detection (IPAD) showed that the decrease in GABA levels (ca. 10% after heating for 4 h) as a function of heating time was smaller than that of glucose (ca. 30% after heating for 4 h). At the same time, glucose to fructose isomerisation took place. After 20 min of heating at pH 6.0, all mixtures showed a fructose peak, the area of which increased with heating time. However, after correcting for fructose isomerisation, glucose losses were still higher than amino acid losses. In contrast to its precursor glutamic acid, GABA was stable during heating of a solution containing it alone. Heating of GABA-containing d-sugar solutions (xylose, fructose, glucose, maltose and sucrose) showed that the relative order of colour yield was pentose > hexose > disaccharides. As well as glucose to fructose isomerisation, HPAEC–IPAD allowed monitoring of the different isomerisation reactions occurring, and also disaccharide hydrolysis in the different GABA/sugar mixtures.     

Oxidative stability enhancement of broiler bird meats with α-lipoic acid and α-tocopherol acetate supplemented feed

Depression of meat quality is known to be caused by lipid peroxidation occurring in meat. Supplementation of antioxidants in feed decreases lipid peroxidation and improves the oxidative stability of meat after slaughtering. The present study demonstrated that meat obtained from broiler birds fed feed supplemented with α-tocopherol acetate (200 mg/kg feed) along with α-lipoic acid (25, 75, or 150 mg/kg feed) exhibited increased oxidative stability and reduced fat content. The total phenolic content and α-lipoic acid content increased in the meat as the concentration of α-lipoic acid supplementation increased. The protein content in the meat was not changed by the supplementation of α-lipoic acid and α-tocopherol acetate. The results of DPPH and TBA assays demonstrated that feed supplemented with α-lipoic acid and α-tocopherol acetate also enhanced the antioxidant activity of broiler meat. On the other hand, the meat from broiler birds fed feed supplemented with oxidised oil (4% in feed) reduced its oxidative stability.     

In vitro α-glucosidase and pancreatic lipase inhibitory activities and antioxidants of Samnamul (Aruncus dioicus) during rehydration and cooking

In vitro α-glucosidase and pancreatic lipase inhibitory activities of ethanol extracts of samnamul (dried young shoots of Aruncus dioicus) during rehydration (soaking, 30 min boiling, and re-soaking in water) and subsequent cooking at 180°C with/without perilla oil were determined for evaluation of its anti-diabetic and antiobesity potential. Polyphenol and flavonoid contents were monitored by spectrophotometry. Rehydration and boiling of samnamul reduced α-glucosidase and pancreatic lipase inhibitory activities as well as polyphenol and flavonoid contents, whereas soaking time did not have a significant effect. Of the rehydration and cooking processes, 30 min boiling was the most critical in reducing anti-diabetic and anti-obesity activities as well as antioxidant contents. Perilla oil addition improved α-glucosidase inhibitory activity, possibly through polyphenol addition. These results suggest that dietary intake of samnamul can reduce the risks of diabetes and obesity, and perilla oil can additionally improve its health benefits.     

Isolation of caffeic acid from Perilla frutescens and its role in enhancing γ-glutamylcysteine synthetase activity and glutathione level

Perilla frutescens is an annual herbaceous plant native to Asia, where its leaves are used in Asian gourmet food. Our previous study showed that the inhibition of γ-glutamylcysteine synthetase (γ-GCS) activity was remarkably recovered by pretreatment with perilla leaf extract (PLE). The objective was to fractionise PLE, and to identify the active component that is responsible for the enhancement of γ-GCS activity and glutathione (GSH) concentration. Among the five fractions from PLE, PLE-III of the ethyl acetate fraction showed the highest γ-GCS activity in a HepG2 cell experiment, and was further chromatographed. The purified compound, which enhanced γ-GCS activity, was finally identified as caffeic acid. We first report the enhancement of γ-GCS activity and GSH level in HepG2 cells by caffeic acid obtained from PLE. Our results suggest that caffeic acid may be a key factor in the chemopreventive potential of perilla leaf components by increasing de novo synthesis of GSH.     

Identification and characterization of novel α-amylase and α-glucosidase inhibitory peptides from camel whey proteins

This study explores the inhibitory properties of camel whey protein hydrolysates (CWPH) toward α-amylase (AAM) and α-glucosidase (AG). A general full factorial design (3 × 3) was applied to study the effect of temperature (30, 37, and 45°C), time (120, 240, and 360 min), and enzyme (pepsin) concentration (E%; 0.5, 1, and 2%). The results showed that maximum degree of hydrolysis was obtained when hydrolysis was carried out at higher temperature (45°C; P < 0.05), compared with lower temperatures of 30 and 37°C. Electrophoretic pattern displays degradation of all protein bands upon hydrolysis by pepsin at various hydrolysis conditions applied. All the 27 CWPH generated showed significant AAM and AG inhibitory potential as indicated by their lower IC₅₀ values (mg/mL) compared with intact whey proteins. In total 196 peptides were identified from selected hydrolysates and 15 potential peptides (PepSite score > 0.8; http://pepsite2.russelllab.org/) were explored via in silico approach. Novel peptides PAGNFLMNGLMHR, PAVACCLPPLPCHM, MLPLMLPFTMGY, and PAGNFLPPVAAAPVM were identified as potential inhibitors for both AAM and AG due to their high number of binding sites and highest binding probability toward the target enzymes. CCGM and MFE, as well as FCCLGPVPP were identified as AG and AAM inhibitory peptides, respectively. This is the first study that reports novel AG and AAM inhibitory peptides from camel whey proteins. The future direction for this research involves synthesis of these potential AG and AAM inhibitory peptides in a pure form and investigate their antidiabetic properties in the in vitro, as well as in vivo models. Thus, CWPH can be considered for potential applications in glycaemic regulation.     

Comparative evaluation of rosmarinic acid,methyl rosmarinate and pedalitin isolated from Rabdosia serra (MAXIM.) HARA as inhibitors of tyrosinase and α-glucosidase

Rabdosia serra has been used in traditional Chinese medicine for centuries. In order to illustrate the pharmaceutical activity of R. serra as hypoglycaemic and skin-whitening agents, rosmarinic acid (confirmed as the major compound in R. serra), methyl rosmarinate and pedalitin isolated from R. serra were evaluated for their inhibitory effects and mechanisms on tyrosinase and α-glucosidase. The inhibitory effects on both tyrosinase and α-glucosidase were in decreasing order, pedalitin > methyl rosmarinate > rosmarinic acid. The IC₅₀ values for the tyrosinase and α-glucosidase activity inhibited by pedalitin were 0.28 and 0.29 mM, respectively. Both rosmarinic acid and methyl rosmarinate were considered as noncompetitive inhibitors of tyrosinase, while pedalitin was suggested to be a mixed-type inhibitor of tyrosinase. In the assay of α-glucosidase inhibition, rosmarinic acid was found to be a competitive inhibitor, whereas both methyl rosmarinate and pedalitin were considered as mixed-type inhibitors.     

Implementation of response surface methodology to assess the antiradical behaviour in mixtures of ascorbic acid and α-tocopherol with grape (Vitis vinifera) stem extracts

The efficiency of grape stem extracts to express antiradical activity was assessed using two different approaches and DPPH as the radical probe. In addition, the mixture effects when the extracts were combined with ascorbic acid (AA) and α-tocopherol (α-Tcp) were also evaluated. The approaches included a simple linear regression analysis between the response (antiradical activity) and concentration, but also a response surface methodology, which permitted the monitoring of the response upon simultaneous variation of both the concentration of the total polyphenols of the extracts and either of the antioxidants (AA and α-Tcp). The deployment of linear regression poses important constraints with regard to concentration ranges, whereas response surface methodology might be a valuable statistical tool for similar assessments and credible modelling of binary mixtures of antioxidants. In all combinations tested it was found that an antagonism was manifested, presumably as a result of AA and α-Tcp regeneration by the extract polyphenols, at the expense of the latter.     

Inhibitory effect of polyphenol-rich extracts of jute leaf (Corchorus olitorius) on key enzyme linked to type 2 diabetes (α-amylase and α-glucosidase) and hypertension (angiotensin I converting) in vitro

Corchorus olitorius leaf is consumed in various parts of the world as leafy vegetable and folk remedy for the management of some degenerative diseases with dearth of information on its biochemical rationale. Therefore, this study sought to characterize the inhibitory action of polyphenol-rich extracts (free and bound) of C. olitorius on α-amylase, α-glucosidase and angiotensin I converting enzyme (ACE), as well as to identify the phenolic compound responsible for these activities. Our findings revealed that the extracts inhibited α-amylase and α-glucosidase (12.5–50.0 μg/mL), and ACE (10.0–50.0 μg/mL) in dose-dependently with free extracts having significantly (P < 0.05) higher α-amylase (17.5 μg/mL), α-glucosidase (11.4 μg/mL) and ACE (15.7 μg/mL) inhibitory activities as revealed by the IC₅₀. Reversed-phase HPLC analysis of the extracts revealed chlorogenic acid (7.5 mg/100 g) and isorhamnetin (51.1 mg/100 g) as the main phenolics in the free extract and caffeic acid (58.1 mg/100 g) in the bound extract. Therefore, the enzyme inhibitory activity of C. olitorius extracts may be attributed to the presence of caffeic acid, chlorogenic acid and isorhamnetin, thus justifying its use in folklore for the management of diabetes and hypertension.