Preparation of fermented sugar-soaked black soybean snacks (FSBSS) and characterization of their quality changes

Fermented sugar-soaked black soybean snacks (FSBSS) were prepared using optimized conditions at 2 times sugar soaking process to give a desirable form and texture after fermentation with Lactobacillus plantarum (LP) and Bacillus subtilis (BS) for 48 h. The prepared FSBSS showed higher total polyphenol and flavonoid contents than an unfermented (NF) sample; total isoflavone content increased approximately 31 and 17% (LP and BS, respectively), as compared to NF. γ-Aminobutyric acid was also contributed by LP and BS, indicating significantly higher contents of 16.9 and 21.6 mg/g (LP and BS, respectively) compared to NF (15.2 mg/g). Moreover, the FSBSS showed antioxidant activities on DPPH, superoxide, and hydroxyl radicals as compared to NF. Based on the results, the newly developed FSBSS using solid-fermentation indicate a useful and convenient way to replace traditional soybean products from the standpoint that also provides an easy means to supply high nutritive and functional properties.     

Angiotensin-converting enzyme inhibitory activity of Lactobacillus helveticus strains from traditional fermented dairy foods and antihypertensive effect of fermented milk of strain H9

Hypertension is a major global health issue which elevates the risk of a large world population to chronic life-threatening diseases. The inhibition of angiotensin-converting enzyme (ACE) is an effective target to manage essential hypertension. In this study, the fermentation properties (titratable acidity, free amino nitrogen, and fermentation time) and ACE-inhibitory (ACEI) activity of fermented milks produced by 259 Lactobacillus helveticus strains previously isolated from traditional Chinese and Mongolian fermented foods were determined. Among them, 37 strains had an ACEI activity of over 50%. The concentrations of the antihypertensive peptides, Ile-Pro-Pro and Val-Pro-Pro, were further determined by ultra performance liquid chromatography with quadrupole-time-of-flight mass spectrometry. The change of ACEI activity of the fermented milks of 3 strains exhibiting the highest ACEI activity upon gastrointestinal protease treatment was assayed. Fermented milks produced by strain H9 (IMAU60208) had the highest in vitro ACEI activity (86.4 ± 1.5%), relatively short fermentation time (7.5 h), and detectable Val-Pro-Pro (2.409 ± 0.229 µM) and Ile-Pro-Pro (1.612 ± 0.114 µM) concentrations. Compared with the control, a single oral dose of H9-fermented milk significantly attenuated the systolic, diastolic, and mean blood pressure of spontaneously hypertensive rats (SHR) by 15 to 18 mmHg during the 6 to 12 h after treatment. The long-term daily H9-fermented milk intake over 7 wk exerted significant antihypertensive effect to SHR, but not normotensive rats, and the systolic and diastolic blood pressure were significantly lower, by 12 and 10 mmHg, respectively, compared with the control receiving saline. The feeding of H9-fermented milk to SHR resulted in a significantly higher weight gain at wk 7 compared with groups receiving saline, commercial yogurt, and captopril. Our study identified a novel probiotic L. helveticus strain originated from kurut sampled from Tibet (China), which is a valuable resource for future development of functional foods for hypertension management.     

Angiotensin-converting enzyme inhibitory activity of milk fermented by wild and industrial Lactococcus lactis strains

Angiotensin I-converting enzyme inhibitory (ACEI) activity was evaluated and compared in <3 KDa water-soluble extracts (WSE) isolated from milk fermented by wild and commercial starter culture Lactococcus lactis strains after 48 h of incubation. The highest ACEI activities were found in WSE from milk inoculated with wild L. lactis strains isolated from artisanal dairy products and commercial starter cultures. On the other hand, the lowest ACEI activities were found in WSE from milk inoculated with wild strains isolated from vegetables. Moreover, the IC₅₀ values (concentration that inhibits 50% activity) of WSE from artisanal dairy products were the lowest, indicating that these fractions were the most effective in inhibiting 50% of ACE activity. In fact, a strain isolated from artisanal cheese presented the lowest IC₅₀ (13 μg/mL). Thus, it appears that wild L. lactis strains isolated from artisanal dairy products and commercial starter cultures showed good potential for the production of fermented dairy products with ACEI properties.     

Functional milk beverage fortified with phenolic compounds extracted from olive vegetation water, and fermented with functional lactic acid bacteria

Functional milk beverages (FMB100 and FMB200) fortified with phenolic compounds (100 and 200 mg/l) extracted from olive vegetable water, and fermented with γ-amino butyric acid (GABA)-producing (Lactobacillus plantarum C48) and autochthonous human gastro-intestinal (Lactobacillus paracasei 15N) lactic acid bacteria were manufactured. A milk beverage (MB), without addition of phenolic compounds, was used as the control. Except for a longer latency phase of FMB200, the three beverages showed an almost similar kinetic of acidification, consumption of lactose and synthesis of lactic acid. Apart from the beverage, Lb. plantarum C48 showed a decrease of ca. Log 2.52-2.24 cfu/ml during storage. The cell density of functional Lb. paracasei 15N remained always above the value of Log 8.0 cfu/ml. During fermentation, the total concentration of free amino acids markedly increased without significant (P > 0.05) differences between beverages. The concentration of GABA increased during fermentation and further storage (63.0 ± 0.6-67.0 ± 2.1 mg/l) without significant (P > 0.05) differences between beverages. After fermentation, FMB100 and FMB200 showed the same phenolic composition of the phenol extract from olive vegetable water but a different ratio between 3,4-DHPEA and 3,4-DHPEA-EDA. During storage, the concentrations of 3,4-DHPEA-EDA, p-HPEA and verbascoside of both FMB100 and FMB200 decreased. Only the concentration of 3,4-DHPEA increased. As shown by SPME-GC-MS analysis, diactetyl, acetoin and, especially, acetaldehyde were the main volatile compounds found. The concentration of phenolic compounds does not interfere with the volatile composition. Sensory analyses based on triangle and paired comparison tests showed that phenolic compounds at the concentrations of 100 or 200 mg/l were suitable for addition to functional milk beverages.     

Optimization of culture condition for ACEI and GABA production by lactic acid bacteria

Abstract: Gamma‐aminobutyric acid (GABA) and angiotensin‐converting enzyme inhibitor (ACEI) are compounds which can influence hypertension. The goal of this study is to optimize the culture condition for GABA and ACEI production by Lactobacillus plantarum NTU 102 fermented skim milk. In this study, we used 3‐factor‐3‐level Box–Behnken design combining with response surface methodology, where the 3 factors represent the concentration of skim milk, the concentration of monosodium glutamate, and culture temperature. Best conditions for GABA and ACEI production differed. The results indicated that L. plantarum NTU 102 produced the highest combined levels of GABA and ACEI at 37 °C, in milk having 8% to 12% nonfat solids supplemented with 0.6% to 1% MSG. Agitation of the medium during fermentation had no effect on GABA or ACEI production but extended incubation (up to 6 d) increases levels of the bioactive compounds. L. plantarum NTU 102 fermented products may be a potential functional food source for regulating hypertension.     

The impact of fermentation and in vitro digestion on formation angiotensin converting enzyme (ACE) inhibitory peptides from pea proteins

Pea seeds were fermented by Lactobacillus plantarum 299v in monoculture under different time and temperature conditions and the fermented products were digested in vitro under gastrointestinal conditions. After fermentation and digestion ACE inhibitory activity was determined. In all samples after fermentation no ACE inhibitory activity was noted. Potentially antihypertensive peptides were released during in vitro digestion. The highest DH (68.62%) were noted for control sample, although the lowest IC₅₀ value (0.19 mg/ml) was determined for product after 7 days fermentation at 22 °C. The hydrolysate characterised by the highest ACE inhibitory activity was separated on Sephadex G10 and two peptides fractions were obtained. The highest ACE inhibitory activity (IC₅₀ = 64.04 μg/ml) for the first fraction was noted. This fraction was separated by HPLC and identified by LC–MS/MS and the sequence of peptide derived from pea proteins was determined as KEDDEEEEQGEEE.     

Angiotensin-converting enzyme inhibitory activity in Mexican Fresco cheese

The objective of this study was to evaluate if Mexican Fresco cheese manufactured with specific lactic acid bacteria (LAB) presented angiotensin I-converting enzyme inhibitory (ACEI) activity. Water-soluble extracts (3 kDa) obtained from Mexican Fresco cheese prepared with specific LAB (Lactococcus, Lactobacillus, Enterococcus, and mixtures: Lactococcus-Lactobacillus and Lactococcus-Enterococcus) were evaluated for ACEI activity. Specific peptide fractions with high ACEI were analyzed using reverse phase-HPLC coupled to mass spectrometry for determination of amino acid sequence. Cheese containing Enterococcus faecium or a Lactococcus lactis ssp. lactis-Enterococcus faecium mixture showed the largest number of fractions with ACEI activity and the lowest half-maximal inhibitory concentration (IC₅₀; <10 μg/mL). Various ACEI peptides derived from β-casein [(f(193-205), f(193-207), and f(193-209)] and α_S1-casein [f(1-15), f(1-22), f(14-23), and f(24-34)] were found. The Mexican Fresco cheese manufactured with specific LAB strains produced peptides with potential antihypertensive activity.     

The antioxidant and free-radical scavenging activities of extract and fractions from corn silk (Zea mays L.) and related flavone glycosides

This study was designed to evaluate both the antioxidant and free-radical scavenging activities of extract and fractions from corn silk. N-butanol fraction (BF) demonstrated the highest total phenolic content (164.1 ± 9.7 μg GAE/g DCS) and total flavonoids content (69.4 ± 5.1 μg RE/g DCS), accompanied with the highest antioxidant activity compared to other fractions through all antioxidant assays. Two flavone glycosides showing potent antioxidant activity were isolated from BF and identified, by comparing spectral data (UV, FAB-MS and NMR) with literature values, to be isoorientin-2″-O-α-l-rhamnoside and 3′-methoxymaysin. The two isolated flavone glycosides, particularly isoorientin-2″-O-α-l-rhamnoside, demonstrated significant total antioxidant activity, DPPH radical scavenging activity, reducing power and iron-chelating capacity, with EC₅₀ values of 14.24 ± 1.49, 22.69 ± 2.33, 6.58 ± 1.07 and 30.25 ± 3.05 μg/ml, respectively. Results obtained indicated that corn silk extracts can be used potentially as a ready accessible and valuable bioactive source of natural antioxidants.     

Casein hydrolysis by Bifidobacterium longum KACC91563 and antioxidant activities of peptides derived therefrom

Milk protein is a well-known precursor protein for the generation of bioactive peptides using lactic acid bacteria. This study investigated the antioxidant activity of bovine casein hydrolysate after fermentation with Bifidobacterium longum KACC91563 using the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay and total phenolic content (TPC). The antioxidant activities of the 24-h and 48-h hydrolysates were higher than that of the 4-h hydrolysate (2,045.5 and 1,629.3 μM gallic acid equivalents, respectively, vs. 40.3 μM) in the ABTS assay. In contrast, TPC values showed activities of 43.2 and 52.4 μM gallic acid equivalents for the 4-h and 24-h hydrolysates, respectively. Three fractions (≥10 kDa, ≥3 but <10 kDa, and <3 kDa) were separated from the 24-h hydrolysate by ultrafiltration. Among these fractions, the <3 kDa fraction exhibited the highest antioxidant activity (936.7 μM) compared with the other fractions (42.1 and 34.2 μM for >10 kDa and 3–10 kDa fractions, respectively). Through liquid chromatography-electrospray ionization-tandem mass spectrometry analysis, 2 peptides, VLSLSQSKVLPVPQK and VLSLSQSKVLPVPQKAVPYPQRDMPIQA, containing the fragment VLPVPQ that has antioxidant properties, were identified in the <3 kDa fraction after 24 h of hydrolysis. The present study demonstrates the possibility of antioxidant peptide production from bovine casein using Bifidobacterium longum.     

Fermented goats’ milk produced with selected multiple starters as a potentially functional food

A screening among five lactic acid bacteria, used alone or in combination, led to select a mixed starter (Streptococcus thermophilus CR12, Lactobacillus casei LC01, Lactobacillus helveticus PR4, Lactobacillus plantarum 1288) capable to produce a fermented goats’ milk containing γ-aminobutyric acid (GABA) and angiotensin-I converting enzyme (ACE)-inhibitory peptides. The fermented milk was characterized by cell counts of lactic acid bacteria not lower than 7.0 log cfu g⁻¹, even after 45 days of storage at 4 °C. Fermentation of goats’ milk resulted in the production of ca. 28 mg kg⁻¹ of GABA. Furthermore the fermented goats’ milk had an in vitro ACE-inhibitory activity of ca. 73%. Prolonged cold storage did not significantly affect both the concentration of GABA and the ACE-inhibitory activity. Moreover, the taurine content did not significantly vary during both fermentation and the entire storage period.     

Optimization of sourdough process for improved sensory profile and texture of wheat bread

The aim of the study was to determine optimum sourdough process conditions for improved flavour and texture of wheat bread. The influence of process conditions and the starter culture on the characteristics of wheat sourdough bread was established by using response surface methodology. Influence of fermentation temperature (16-32 °C), ash content of flour (0.6-1.8 g/100 g), and fermentation time (6-20 h) were considered as independent factors and their effects were studied in sourdough bread fermented with Lactobacillus plantarum, Lactobacillus brevis, Saccharomyces cerevisiae or with a combination of yeast and lactic acid bacteria. Intensity of sensory attributes, specific volume and bread hardness were considered as the main responses. Ash content of flour and fermentation time were the main factors determining the intensity of sensory attributes. The possibility to enhance intensity of overall flavour, aftertaste and roasted flavour without excessive pungent flavour and without reduced fresh flavour in wheat bread containing 20 g sourdough/100 g of wheat dough was demonstrated by choosing e.g. Lb. brevis for a starter and by utilization of high ash content of flour, long fermentation time and reduced temperature. Bread specific volume was improved 0.2-0.5 ml/g and hardness was reduced (after 4 days of storage) up to 260 g by using low ash content of flour and by optimizing fermentation time according to the microbial strain. Lactic acid fermentation had more profound influence on both desired and undesired flavour attributes, as well as textural features of bread in comparison with yeast fermentation.     

Study of the behaviour of Lactobacillus plantarum and Leuconostoc starters during a complete wheat sourdough breadmaking process

The acidification properties, metabolic activity and technological performance of four individual Lactobacillus plantarum or Leuconostoc freeze-dried starters were investigated during a complete wheat sourdough breadmaking process including 0.2 g/100 g baker's yeast. Microbiological contents (lactic acid bacteria and yeasts), acidification characteristics (pH and total titratable acidity), soluble carbohydrates (maltose, glucose and fructose) and fermentative end-products (lactic and acetic acids, ethanol) contents were evaluated during both sourdough and corresponding bread dough fermentation. Biochemical and technological analysis of the resulting bread products are also presented. Some differences among strains in acidification properties and soluble carbohydrates availability were outlined both in sourdough and bread dough. Each individual Leuconostoc or Lb. plantarum starter was able to produce a characteristic fermentation and was found to ensure the production of breads with overall satisfactory acceptance.     

Brewing of glucuronic acid-enriched apple cider with enhanced antioxidant activities through the co-fermentation of yeast (Saccharomyces cerevisiae and Pichia kudriavzevii) and bacteria (Lactobacillus plantarum)

Co-fermentation using yeast (Saccharomyces cerevisiae and Pichia kudriavzevii) and the bacteria (Lactobacillus plantarum) as starters isolated from spontaneous sourdough was conducted for the brewing of glucuronic acid (GlcA)-enriched apple cider. The concentration of GlcA in the apple cider co-fermented for 14 d with commercial S. cerevisiae and L. plantarum was 37.7 ± 1.7 mg/mL while a concentration of 62.8 ± 3.1 mg/mL was recorded for fermentation with P. kudriavzevii and L. plantarum, which was higher than the corresponding single yeast fermentation. The co-fermented apple cider revealed higher 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of 171.67 ± 0.79 µg trolox equivalents (TE)/mL using P. kudriavzevii and L. plantarum, compared to the control (143.89 ± 7.07 µg TE/mL) just using S. cerevisiae. Thus, the co-fermentation of S. cerevisiae and L. plantarum and P. kudriavzevii and L. plantarum provided a new strategy for the development of GlcA-enriched apple cider with enhanced antioxidant capacity.     

Production and characterization of a milk-clotting protease in the crude enzymatic extract from the newly isolated Thermomucor indicae-seudaticae N31: (Milk-clotting protease from the newly isolated Thermomucor indicae-seudaticae N31)

Microbial rennet-like milk-clotting enzymes are aspartic proteinases that catalyze milk coagulation, substituting calf rennet. Crude enzymatic extract produced by the thermophilic fungus, Thermomucor indicae-seudaticae N31, on solid state fermentation (SSF) using wheat bran, exhibited high milk-clotting activity and low proteolytic activity after 24 h of fermentation. Highest milk-clotting activity (MCA) was at pH 5.7, at 70 °C and in 0.04 M CaCl₂; it was stable in the pH range 3.5–4.5 for 24 h and up to 45 °C for 1 h. MCA was highly inhibited by pepstatin A. Hydrolytic activity profile of the crude enzymatic extract on whole bovine casein, analyzed by gel electrophoresis (Urea–PAGE) and RP-HPLC revealed low proteolytic action towards casein fractions and a peptide profile similar to the one obtained with commercial Rhizomucor miehei protease (Hannilase).     

Quality protein maize (Zea mays L.) tempeh flour through solid state fermentation process

Solid state fermentation (SSF) process represents a technological alternative for a great variety of legumes and cereals, or combination of them, to improve their nutritional quality and to obtain edible products with palatable sensorial characteristics. The objectives of this work were (1) to determine the best combination of SSF process variables (temperature/time) for production of quality protein maize (Zea mays L.) (QPM) tempeh flour, and (2) to characterize physico-chemical properties and nutritionally quality of optimized QPM tempeh flour. Response surface methodology (RSM) was applied as optimization technique, over three response variables: In vitro protein digestibility (PD), true protein (TD) and water absorption index (WAI). A central composite experimental design with two factors [fermentation temperature (FT), fermentation time (Ft)] and five levels (2 factorial, 2 axial, 1 central) was used. The process variables, FT and Ft, had variation levels of 31-36°C and 48-72 h, respectively. Spores from Rhizopus oligosporus were suspended in distilled water (1×10⁶ spores/ml) and used as starter. Predictive models for response variables were developed as a function of process variables. Conventional graphical method was applied to obtain maximum PD, TP and WAI. Contour plots of each of the response variables were utilized, applying superposition surface methodology, to obtain a contour plot for observation and selection of the best combination of FT (35.4°C) and Ft (54.6 h) for production of QPM tempeh, which was dried (52°C, 24 h) and milled to pass through a 80-US mesh (0.180 mm) screen to obtain optimized QPM tempeh flour. This flour had higher (p?0.05) true protein (13.1 vs 9.1 g/100 g, DM), total colour difference (21.7 vs 9.2), WAI (2.9 vs 1.25 g gel/g dry flour), available lysine (5.67 vs 4.2 g lysine/100 g protein) and PD (83.6 vs 78.5%, DM), and lower (p?0.05) lipid content (4.3 vs 6.1 g/100 g, DM), phytic acid (0.8 vs 10.1 mg phytic acid/g, DM), and pH (4.5 vs 6.12) than raw QPM flour.     

Preparation and antihypertensive activity of peptides from Porphyra yezoensis

This research was to develop a antihypertensive peptide, an efficient angiotensin converting enzyme (ACE) inhibitor (ACEI), from Porphyra yezoensis. Seven commercial enzymes were screened and then enzymatic hydrolysis conditions were optimised. The results showed that alcalase was the most effective for hydrolysis and its optimum conditions for achieving the highest antihypertensive activity of peptide were 1.5% substrate, 5% alcalase, pH 9.0, and temperature of 50 °C at a hydrolysis time of 60 min. The antihypertensive peptide produced under the optimum conditions had a high ACE inhibition rate of 55.0% and a low IC₅₀ value of 1.6 g/l and remained at high stability at temperatures of 4, 25, and 37 °C, pH values of 2.0 and 8.0, and after pepsin and trypsin treatments. Major proteins from P. yezoensis were glutelin, albumin, and gliadin. The albumin and glutelin had higher hydrolysis rates than the gliadin, but the IC₅₀ value of glutelin was the lowest, which indicated that the antihypertensive peptide from glutelin was more functional.     

Influence of alcoholic and malolactic starter cultures on bioactive amines in Merlot wines

The influence of alcoholic and malolactic fermentations on the levels of amines in Merlot wines was investigated. Saccharomyces bayanus, S. cerevisiae, Lactobacillus plantarum, Oenococcus oeni (DSM 7008 and 12923) and spontaneous fermentations were used. Four of the 10 amines investigated were detected: spermidine, serotonin, putrescine and cadaverine. When considering the factors independently, the malolactic bacteria significantly affected the levels of serotonin and total amines, whereas the fermentation yeasts significantly affected the levels of spermidine (two way Kruskal–Wallis, p ? 0.05). Spermidine levels were significantly higher in wines produced with S. cerevisiae. Significantly higher serotonin levels were found in wines made with L. plantarum. Putrescine and cadaverine were not detected in wines produced by spontaneous alcoholic fermentation or by L. plantarum. There were significant differences in alcohol content, total and volatile acidity, sulphite levels and taste quality among wines (Tukey test, p ? 0.05).     

Properties and acceptability of Som-fug, a Thai fermented fish mince, inoculated with lactic acid bacteria starters

Effect of inoculation of different lactic acid bacteria (LAB) on the fermentation and quality of Som-fug from bigeye snapper was investigated. Som-fug inoculated with Pediococcus acidilactici at 10⁴ CFU/g (PA104) had a greater acceptability than those inoculated with Lactobacillus plantarum and Pediococcus pentosaceus at either 10⁴ or 10⁶ CFU/g and the control (without inoculum). During fermentation, PA104 exhibited a higher rate of fermentation than the control as indicated by the greater rate of pH drop and lactic acid production (p<0.05). Based on pH desired (pH 4.5), the fermentation was completed within 48 and 36 h for the control and PA104, respectively. Due to higher acid production rate, the textural development of PA104 was more pronounced. Som-fug inoculated with LAB generally exhibited higher hardness and adhesiveness than the control (p<0.05), when assessed by texture profile analysis (TPA). From the result, inoculation with P. acidilactici resulted in a reduction of fermentation time and improved the quality of Som-fug. Therefore, P. acidilactici can be used as a potential starter for Som-fug fermentation.     

The effects of various antifungal additives on the fermentation and aerobic stability of corn silage

In 2 consecutive years, whole plant corn was ensiled in laboratory silos to investigate the effects of various silage additives on fermentation, dry matter (DM) recovery and aerobic stability. In yr 1, chopped forage was treated with 1) no additive (untreated, U), 2) Lactobacillus buchneri40788, 4 × 10⁵ cfu/g of fresh forage (LLB4), 3) L. buchneri 11A44, 1 × 10⁵ cfu/g (PLB), 4) Biomax 5 (Lactobacillus plantarum PA-28 and K-270), 1 × 10⁵ cfu/g (B5), 5) Silo Guard II (sodium metabisulfite and amylase), 0.05% of fresh forage weight (SG), 6) a buffered propionic acid-based additive, 0.1% (Ki-112), 7), sodium benzoate, 0.1% of fresh weight (SB), or 8) potassium sorbate:EDTA (1:1), 0.1% of fresh weight (PSE). Silage treated with LLB4 had the highest concentration of acetic acid compared with other treatments, and yeasts were undetectable in LLB4 (<log₂ cfu/g). Silages treated with SB and PSE had the highest concentrations of water-soluble carbohydrates, the greatest recoveries of DM, and the lowest concentrations of ethanol. Silages treated with B5, SG, and Ki-112 had no effects on fermentation, DM recovery, or aerobic stability. The aerobic stabilities of silages treated with LLB4, SB, and PSE were greatest among all treatments. In yr 2, treatments were: 1) U, 2) LLB4, 3) PLB, 4) PLB at 4 × 10⁵ cfu/g (PLB4), and 5) B5. Silages treated with L. buchneri had greater concentrations of acetic acid but lower concentrations of ethanol than did U- and B5-treated silages. Yeasts were undetected in all silages except in silage treated with B5, which had the poorest aerobic stability of all treatments. Treatments had no effect on DM recovery. Silages treated with PLB, PLB4, and LLB4 remained stable for >210 h.     

Honey flavonoids as protection agents against oxidative damage to human red blood cells

Honey phenol extracts separated on the base of their hydrophobicity were evaluated for the antioxidant content and for the ability to inhibit oxidative damage induced by radical species generated in the water phase or in the membrane of human erythrocytes. The water and ether fractions obtained from crude methanol extract of honey exhibited a phenolic content of 5.33 and 2.62 mg caffeic acid equivalents/100 g honey, respectively. These values correlate well with those of total antioxidant power, as assessed by FRAP assay (37.67 vs. 10.65 μmol/100 g honey). Flavonoid contents were 2.57 and 1.64 mg catechin equivalents/100 g honey for ether and water fractions, respectively. Although both honey fractions protect erythrocytes against 2,2′-azobis(2-amidinopropane)dihydrochloride-induced lysis, only the ether fraction was found to be active in inhibiting hemolysis but not methemoglobin and ferrylhemoglobin formation caused by H₂O₂. In addition, the ether fraction prevents tert-butylhydroperoxide-induced lipid peroxidation in whole erythrocytes and in isolated membranes. The significant antioxidant effect against damages induced by both water-soluble and hydrophobic exogenous oxidants suggests that the ether fraction, owing to its lipophilic character, can interact with red blood cell membrane, and the protective effect can be associated with the binding of the flavonoids to the membrane. On the other hand, the water fraction is more hydrophilic than ether fraction and it acts only from the outside of the membrane by scavenging the radicals before they attack the erythrocyte membrane.