Antioxidant Capacity of Alcalase Hydrolysates and Protein Profiles of Two Conventional and Seven Low Glycinin Soybean Cultivars

Soy protein hydrolysates are considered a potential dietary source of natural antioxidants with important biological activities. This study was conducted to compare the effect of two conventional and seven low glycinin soybean cultivars on the antioxidant capacity (AC) of soy hydrolysates. Nine cultivars were grown in Bloomington, IL, Findlay, OH and Huxley, IA. The hydrolysates were produced enzymatically using alcalase and analyzed for AC using oxygen radical absorbance capacity (ORAC) assay and soluble protein. Statistical differences were observed in the protein profiles and AC among the different cultivars tested (P < 0.05). The hydrolysate from low glycinin cultivar 3 enriched in β-conglycinin, grown in Bloomington, exhibited the highest AC, compared to the other cultivars across all locations. On average, soy cultivars rich in BC and purified BC hydrolysates (36.2 and 31.8 μM Trolox equivalents (TE)/μg soluble protein, respectively) (P > 0.05) had higher AC than purified glycinin (GL) hydrolysate (28.5 μM TE/μg soluble protein) (P < 0.05). It was possible to select a soybean cultivar that produced a higher antioxidant capacity upon alcalase hydrolysis.     

Organic and Genetically Modified Soybean Diets: Consequences in Growth and in Hematological Indicators of Aged Rats

The aim of this study was to evaluate the protein quality of organic and genetically modified soy by feeding specific diets to rats. Three groups of Wistar rats (n = 10) were used, and each group was named according to the food that they ate. There was an organic soy group (OG), a genetically modified soy group (GG), and a control group (CG). All animals received water and diet ad libitum for 455 days. At the end of this period, the weight of the GG group was the same as that of the OG, and both were higher than CG. Protein intake was similar for the OG and GG, which were significantly lower (p < 0.0005) than the CG. The growth rate (GR) of the rats, albumin levels, and total levels of serum protein were comparable for all groups. Hematocrit (p < 0.04) and hemoglobin (p < 0.03) for the OG and GG were less than the CG. Although the OG and GG demonstrated reduced hematocrit and hemoglobin, both types of soy were utilized in a way similar to casein. This result suggests that the protein quality of soy is parallel to the standard protein casein in terms of growth promotion but not hematological indicators.     

Nutrient Based Estimation of Acid-Base Balance in Vegetarians and Non-vegetarians

A first objective of the present study was to estimate the acid-base balance of the food intake in vegetarians and non-vegetarians. A second objective was to evaluate if additional input of specific food items on the existing potential renal acid load (PRAL) list was necessary for the comparison of the two dietary patterns. Thirty vegetarians between the age of 18 and 30 years were matched for sex, age and BMI with 30 non-vegetarians. Based on the 3-days food diaries the acid-base status of the food intake was estimated using the PRAL method. Mean PRAL values as estimated with the standard table yielded an alkaline load of -5,4 ± 14,4 mEq/d in the vegetarians compared to an acid load of 10,3 ± 14,4 mEq/d in the non-vegetarians (p < 0,001). Mean PRAL values as estimated with the extended table yielded an alkaline load of -10,9 ± 19,7 mEq/d in the vegetarians compared to an acid load of 13,8 ± 17,1 mEq/d for the non-vegetarians (p < 0,001). The findings of this study indicate that vegetarian food intake produces more alkaline outcomes compared to non-vegetarian diets. The use of the standard PRAL table was sufficient for discrimination between the two diets.     

Bioavailability of Phenolic Antioxidants Associated with Dietary Fiber: Plasma Antioxidant Capacity After Acute and Long-Term Intake in Humans

Most studies on bioavailability of phenolic antioxidants are focused in foods and beverages in which they may be easily released from the food matrix, reaching a peak in plasma antioxidant capacity 1–2 h after the intake. However, plant foods contain significant amounts of polyphenols associated with dietary fiber. The aim of the present work was to seek the bioavailability of total phenolic antioxidants associated with dietary fiber by measuring plasma antioxidant capacity in human volunteers. An acute intake of 15 g of a dietary fiber rich in associated phenolic antioxidants in healthy volunteers (n = 10) increased antioxidant capacity of plasma in relation to a control group (n = 4), becoming significant 8 h after the intake. This shows that phenolic antioxidants associated with dietary fiber are at least partially bioavailable in humans, although dietary fiber appears to delay their absorption. No significant changes were observed after long-term intake (16 weeks, 34 subjects).     

Effect of High β-Glucan Barley on Serum Cholesterol Concentrations and Visceral Fat Area in Japanese Men—A Randomized,Double-blinded,Placebo-controlled Trial

This study investigated whether the consumption of a diet in which high-β-glucan barley replaced rice would reduce the visceral fat area as well as the serum low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC) in hypercholesterolemic Japanese men. A randomized, double-blinded, placebo-controlled intervention study was conducted in 44 hypercholesterolemic Japanese men with a body mass index (BMI) >22 kg/m². The subjects were randomly assigned to groups consuming either rice (placebo group) or a mixture of rice and pearl barley with a high β-glucan content (test group, 7.0 g β-glucan per day) for 12 weeks. Blood samples were taken, and CT scan obtained before the trial and every four weeks during the trial. The pearl barley intake significantly reduced serum concentrations of LDL-C (P = 0.041) and TC (P = 0.037) during the trial. Significant differences between the test and placebo groups were found for the visceral fat (P = 0.039), BMI (P = 0.015), and waist circumference (P = 0.011) at the end point. The consumption of pearl barley with a high β-glucan content reduces not only LDL-C but also visceral fat area.     

Effects of enzymatic hydrolysis on molecular structure and antioxidant activity of barley hordein

Hordein, the major storage protein of barley (Hordeum vulgare L.), was hydrolysed by three selected proteases, including alcalase, flavourzyme and pepsin. The effects of protease type and hydrolysis time on hordein molecular weight, surface hydrophobicity, secondary structure and antioxidant activity were investigated. Flavourzyme hydrolysis of hordein was relatively more extensive and rapid, resulting in the formation of medium- and small-sized peptides with a broad distribution within 30 min. Alcalase and pepsin more gradually and less extensively hydrolysed hordein into medium- and larger-sized peptides, respectively. Protein surface hydrophobicity decreased with an increasing degree of hydrolysis. The flavourzyme and alcalase hydrolysates had superior DPPH (1,1-diphenyl-2-picryl hydrazyl) free radical scavenging activity (44-70 and 48-58%, respectively, at 0.5 mg/mL), Fe²⁺-chelating ability (21-64% and 39-73%, respectively, at 1 mg/mL), and superoxide radical scavenging capacity. It is proposed that the large- and medium-size hydrolysate fractions were most likely responsible for the antioxidant activities of hordein hydrolysates, and could be used as antioxidant peptides in food and nutraceutical applications.     

Effects of Different Sources of Fructans on Body Weight,Blood Metabolites and Fecal Bacteria in Normal and Obese non-diabetic and Diabetic Rats

Fructans contribute significantly to dietary fiber with beneficial effects on gastrointestinal physiology in healthy individuals and offer a promising approach to treating some diseases. Two experiments (Experiment 1 = rats with normal weight; Experiment 2 = obese rats) were developed to compare the effects of three fructan sources (Cichorium intybus L. Asteraceae, Helianthus tuberosus L. Asteraceae and Agave angustifolia ssp. tequilana Haw, Agavaceae) on body weight change, blood metabolites and fecal bacteria in non-diabetic (ND) and diabetic (D) rats. In Experiment 1 total body weight gain and daily feed intake in D and ND rats decreased (P < 0.05) with supplements of fructan. Only in D rats, blood glucose concentrations, fecal Clostrodium spp. counts, and liver steatosis decreased, while blood HDL concentrations and fecal Lactobacillus spp. and Bifidobacterium spp. counts increased due to fructans. In Experiment 2, total body weight gain and feed intake in ND and D rats were also decreased by fructans. In ND rats, fructan decreased blood glucose concentrations. In D rats, fructans from A. angustifolia ssp. tequilana decreased blood cholesterol and LDL and liver steatosis. For both ND and D rats, fecal Lactobacillus spp. and Bifidobacterium spp. counts were higher (P < 0.05) with fructan supplements.     

Children Consuming Cassava as a Staple Food are at Risk for Inadequate Zinc, Iron, and Vitamin A Intake

Cassava contains little zinc, iron, and β-carotene, yet it is the primary staple crop of over 250 million Africans. This study used a 24-hour dietary recall to test the hypothesis that among healthy children aged 2–5 years in Nigeria and Kenya, cassava’s contribution to the childrens’ daily diets is inversely related to intakes of zinc, iron, and vitamin A. Dietary and demographic data and anthropometric measurements were collected from 449 Kenyan and 793 Nigerian children. Among Kenyan children 89% derived at least 25% of their dietary energy from cassava, while among the Nigerian children 31% derived at least 25% of energy from cassava. Spearman’s correlation coefficient between the fraction of dietary energy obtained from cassava and vitamin A intake was r = −0.15, P < 0.0001, zinc intake was r = −0.11, P < 0.0001 and iron intake was r = −0.36, P < 0.0001. In Kenya, 59% of children consumed adequate vitamin A, 22% iron, and 31% zinc. In Nigeria, 17% of children had adequate intake of vitamin A, 57% iron, and 41% zinc. Consumption of cassava is a risk factor for inadequate vitamin A, zinc and/or iron intake.     

Insoluble Carob Fiber Rich in Polyphenols Lowers Total and LDL Cholesterol in Hypercholesterolemic Sujects

Recently, polyphenols have been found to affect blood lipids in animals in a similar manner as soluble dietary fibre. The aim was to assess whether an insoluble dietary fiber very rich in polyphenols has a beneficial effect on serum lipids in humans. In a double-blind randomized placebo-controlled clinical study with parallel arms, 88 volunteers with hypercholesterolemia were randomly assigned to consume daily either, fiber with insoluble 84% polyphenols 4 g twice a day (n = 43) or placebo (n = 45). Serum total, LDL and HDL cholesterol and triglycerides were assessed at baseline and after 4 weeks. The insoluble polyphenols consumption reduced the total cholesterol by 17.8 ± 6.1% (p < 0.05), LDL cholesterol by 22.5 ± 8.9% (p < 0.001), LDL: HDL cholesterol ratio by 26.2 ± 14.3% (p < 0.001) and triglycerides by 16.3 ± 23.4% (p < 0.05) at the end of the study compared with baseline. No significant differences were found during the study time in the placebo group for the lipid profile. The consumption of fiber very rich in insoluble polyphenols shows beneficial effects on human blood lipid profile and may be effective in prevention and treatment of hyperlipemia.     

Circulating Salicylic Acid and Metabolic and Inflammatory Responses after Fruit Ingestion

We hypothesized that fruit ingestion provides measurable amounts of salicylic acid (SA) and produces different metabolic and inflammatory responses compared to mere fruit sugars. In a randomized-crossover study, 26 healthy subjects received a peach shake meal (PSM) (SA: 0,06 ± 0,001 mg/100 g) and a mixed sugar meal (MSM), consisting in an aqueous solution with the same sugars found in the peach shake. In order to control for the SA contribution from meals in the previous day, 16 subjects (Group 1) abstained from fruits and vegetables consumption the evening before trials, and 10 subjects (Group 2) maintained their usual diet. Circulating SA, glucose, insulin, free fatty acids, and interleukin-6 were determined. Basal SA was lower in Group 1 than in Group 2 (0.09 ± 0.02 vs. 0.30 ± 0.03 μmol/l, p < 0.001), peaked at 90 min in both groups (0.18 ± 0.01 vs. 0.38 ± 0.02 μmol/l, p < 0.01) and remained above baseline (p < 0.05) up to 3 h. Glycemia increased less after PSM at 15 min (p < 0.01) with a lower average glucose excursion (p < 0.05). Insulin peaked at 45 min with both meals but decreased less rapidly with PSM. Free fatty acids decreased more (p < 0.01), and interleukin-6 increased less (p < 0.05) with PSM. Dietary fruit intake increases the concentration of SA in vivo, and provides non-nutrients capable to modulate the inflammatory and metabolic responses to carbohydrates.     

The Effect of Consumption of Selenium Enriched Rye/Wheat Sourdough Bread on the Body’s Selenium Status

The potential of selenium-enriched rye/wheat sourdough bread as a route for supplementing dietary selenium intakes is reported. In addition to their normal diets, 24 female volunteers (24 to 25 years old) were fed either selenium-enriched bread or non-enriched bread each day (68.02 and 0.84 μg selenium day⁻¹ respectively) for 4 weeks. The chemical form of the selenium in the bread had been characterised using HPLC-ICP-MS, which showed that 42% of the extractable selenium was present as selenomethionine. Plasma selenium levels and plasma platelet glutathione peroxidase (GPx1) activity were measured in the volunteers’ blood over a 6-week period. A statistically significant difference (p = 0.001) was observed in the mean percentage change data, calculated from the plasma selenium level measurements for the enriched and control group, over the duration of the study. A comparable difference was not observed for the platelet GPx1 activity (p = 0.756), over the same period. Two weeks after cessation of the feeding stage, i.e., at t = 6 weeks, the mean percentage change value for the selenium plasma levels in the enriched group was still significantly elevated, suggesting that the absorbed selenium had been incorporated into the body’s selenium reserves, and was then being slowly released back into the volunteers’ blood.     

Physicochemical,Nutritional and Antibacterial Characteristics of the Fruit of <Emphasis Type="Italic">Bromelia pinguin</Emphasis> L.

Bromelia pinguin L. is a plant native to Sinaloa, Mexico, where its fruit is used as food or as a phytotherapeutic agent. The fruits of B. pinguin were characterized and they could be considered as a functional food. These fruits show an average weight of 13.7 g and a yellow color of high luminosity (b* = 43.2, L* = 74.5). The values for acidity (4.6%, as citric acid) and pH (3.7) of B. pinguin fruit are similar to those of citrics. The edible portion is characterized by a high content of vitamin C (126 mg/100 g), ash (10.6 g/100 g d.w.), crude fiber (3.4 g/100 g d.w.), calcium (1290 mg/100 g d.w.), magnesium (500 mg/100 g d.w.), manganese (2.95 mg/100 g d.w.) and it is a good source of zinc (2.8 mg/100 g d.w.). Polar fractions extracted from the pulp fruit showed activity against several genera of human pathogenic-bacteria (Staphylococcus, Streptococcus, Enterococcus, Salmonella, Shigella, Escherichia and Pseudomonas).     

Comparison of the Nutrient and Chemical Contents of Traditional Korean Chungtaejeon and Green Teas

This study was conducted in order to compare the nutrient and chemical contents of two Korean teas: traditional Chungtaejeon tea (CTJ) with that of green tea (GT). Main bioactive compounds and the antioxidant activities using four radical scavenging assays (ABTS, CUPRAC, FRAP and DPPH) in methanol and acetone extracts of both teas were determined. It was found that the contents of vitamin C, amino acids and total nitrogen in CTJ were lower than that of GT (p < 0.05). Caffeine, reducing sugar and chlorophyll contents in CTJ were similar to GT. Catechin (C), epicatechin (EC), and epigallocatechin (EGC) contents were lower in CTJ than in green tea (p < 0.05), but gallocatechin (GC), epicatechin gallate (ECG), epigallocatechin gallate (EGCG) and catechin gallate (CG) showed no significant differences between CTJ and GT (p ≥ 0.05). The contents of polyphenols, flavonoids, flavanols and tannins in CTJ methanol were 229.30 ± 11.3 mg GAE/g dry weight (DW), 15.24 ± 0.8 mg CE/g DW, 109.10 ± 5.1 mg CE/g DW and 25.68 ± 1.2 mg CE/g DW, respectively, and significantly higher than in acetone extracts (p < 0.05). Flavonoids (quercetin and kaempferol) were higher in GT than in CTJ and myricetin was higher in CTJ (p < 0.05). Threonine and aspartic acid was lower, and glutamic acid was higher in CTJ compared with GT (p < 0.05). Free amino acid content in CTJ and GT showed no significant difference. Potassium and magnesium in CTJ were lower compared to GT, but no significant difference was found for iron, manganese and calcium. Also, the level of the antioxidant activity by all four used assays was significantly higher in CTJ and in methanol was higher than in acetone extracts (p < 0.05). In conclusion, traditional fermented Korean tea Chungtaejeon contains high quantities of bioactive compounds and possesses high antioxidant activity. The contents of the bioactive compounds and the levels of antioxidant activities are significantly higher in methanol than in acetone extracts. 3-D fluorescence and FTIR- spectroscopy showed slight differences between the two investigated tea samples and can be used as additional tools for identification of polyphenols. Both studied teas can be recommended as a source of bioactive compounds.     

Effect of Quinoa Seeds (Chenopodium quinoa) in Diet on some Biochemical Parameters and Essential Elements in Blood of High Fructose-Fed Rats

The effect of Chenopodium quinoa seeds on lipid profile, glucose level, protein metabolism and selected essential elements (Na, K, Ca, Mg) level was determined in high—fructose fed male Wistar rats. Fructose decreased significantly LDL [42%, p < 0.01] and activity of alkaline phosphatase [20%, p < 0.05], and increased triglycerides level [86%, p < 0.01]. The analysis of blood of rats fed quinoa indicated, that these seeds effectively reduced serum total cholesterol [26%, p < 0.05], LDL [57%, p < 0.008] and triglycerides [11%, p < 0.05] when compared to the control group. Quinoa seeds also significantly reduced the level of glucose [10%, p < 0.01] and plasma total protein level [16%, p < 0.001]. Fructose significantly decreased HDL [15%, p < 0.05] level in control group but when the quinoa seeds were added into the diet the decrease of HDL level was inhibited. Quinoa seeds did not prevent any adverse effect of increasing triglyceride level caused by fructose. It was shown in this study that quinoa seeds can reduce most of the adverse effects exerted by fructose on lipid profile and glucose level.     

Effect of Green Tea and Vitamin E Combination in Isoproterenol Induced Myocardial Infarction in Rats

The present study was aimed to investigate the combined effects of green tea and vitamin E on heart weight, body weight, serum marker enzymes, lipid peroxidation, endogenous antioxidants and membrane bound ATPases in isoproterenol (ISO)-induced myocardial infarction in rats. Adult male albino rats, treated with ISO (200 mg/kg, s.c.) for 2 days at an interval of 24 h caused a significant (P < 0.05) elevation of heart weight, serum marker enzymes, lipid peroxidation and Ca⁺² ATPase level whereas there was a significant (P < 0.05) decrease in body weight, endogenous antioxidants, Na⁺/ K⁺ ATPase and Mg⁺² ATPase levels. Administration of green tea (100 mg/kg/day, p.o.) and vitamin E (100 mg/kg/day, p.o.) together for 30 consecutive days and challenged with ISO on the day 29th and 30th, showed a significant (P < 0.05) decrease in heart weight, serum marker enzymes, lipid peroxidation, Ca⁺² ATPase and a significant increase in the body weight, endogenous antioxidants, Na⁺/K⁺ ATPase and Mg⁺² ATPase when compared with ISO treated group and green tea or vitamin E alone treated groups. These findings indicate the synergistic protective effect of green tea and vitamin E during ISO induced myocardial infarction in rats.     

Combination of On-Line Solid-Phase Extraction with LC-MS for the Determination of Potentially Hazardous Glycoalkaloids in Potato Products

A simple and rapid method for the determination of naturally occurring, potentially hazardous glycoalkaloids (GAs) in potatoes and their products has been developed. The procedure is based on the on-line solid-phase extraction of the acetic acid extracts from potato products and combined with liquid chromatography (LC)-mass spectrometry (MS) in a fully automated system (Symbiosis™, Spark Holland Instruments, Emmen, The Netherlands). As sorbent material HySphere™ 18HD was used for alkaloid enrichment. GAs were eluted with the LC gradient and directly analysed by MS. Detection of the analytes was achieved in the sensitive multiple reaction monitoring mode using two characteristic ions (m/z 98 as a qualifier for GAs and m/z 868.3 as a quantifier for α-solanine or m/z 852.4 for α-chaconine). Typical validation data for method precision (v _k α-solanine = 5.3–6.5, v _k α-chaconine = 3.4–15.4), accuracy (average recovery of α-solanine = 84%, average recovery of α-chaconine = 87%) and linearity over the range from 1 to 1,000 ng ml⁻¹ (R ² = 0.9915 for α-solanine, R ² = 0.9939 for α-chaconine) with detection limits of 0.3 ng ml⁻¹ for α-chaconine and 0.5 ng ml⁻¹ for α-solanine were obtained. GA contents of commercial potato products were determined by the new on-line method and afterwards compared with those obtained with an established high-performance LC routine procedure. Better performance of the on-line procedure was obvious from the standard deviations of both methods. Other advantages included a strong reduction of overall analysis time, human intervention and solvent consumption as well as waste production. The time required for the on-line analysis was 5 min, which would allow processing of almost 100 samples in 8 h.     

Sunflower Protein Hydrolysates Reduce Cholesterol Micellar Solubility

Plant protein hydrolysates are a source of bioactive peptides. There are peptides that decrease the micellar cholesterol solubility from bile acids and therefore may reduce in vivo cholesterol absorption. The presence of these peptides in sunflower protein hydrolysates has been studied. Sunflower protein hydrolysates produced with alcalase plus flavourzyme or with pepsin plus pancreatin inhibited in some degree the cholesterol incorporation to micelles. Protein hydrolysates generated after 30 min of hydrolysis with alcalase, and after 30 min of hydrolysis with pepsin, were the inhibitoriest of the cholesterol incorporation to micelles. The average amino acid hydrophobicity of inhibitory peptides in cholesterol micelles was higher than the observed in the corresponding protein hydrolysates. This high hydrophobicity probably favours their inclusion in the lipid micelles. In vivo, this inhibition may translate in a decrease of cholesterol absorption. Reported results show that a combination of different characteristics such as peptide size or hydrophobicity may be responsible of the inhibitory activity of generated peptides.     

Purification of Antioxidant Peptides by High Resolution Mass Spectrometry from Simulated Gastrointestinal Digestion Hydrolysates of Alaska Pollock (Theragra chalcogramma) Skin Collagen

In this study, the stable collagen hydrolysate was prepared by alcalase hydrolysis and twice simulated gastrointestinal digestion from Alaska pollock skin. The characteristics of hydrolysates and antioxidant activities in vitro, including 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical (ABTS^•+) scavenging activity, ferric-reducing antioxidant power (FRAP) and hydroxyl radical (OH·) scavenging activity, were determined. After twice simulated gastrointestinal digestion of skin collagen (SGI-2), the degree of hydrolysis (DH) reached 26.17%. The main molecular weight fractions of SGI-2 were 1026.26 and 640.53 Da, accounting for 59.49% and 18.34%, respectively. Amino acid composition analysis showed that SGI-2 had high content of total hydrophobic amino acid (307.98/1000). With the simulated gastrointestinal digestion progressing, the antioxidant activities increased significantly (p < 0.05). SGI-2 was further purified by gel filtration chromatography, ion exchange chromatography and high performance liquid chromatography, and the A_1a3c–p fraction with high hydroxyl radical scavenging activity (IC50 = 7.63 μg/mL) was obtained. The molecular weights and amino acid sequences of key peptides of A_1a3c–p were analyzed using high resolution mass spectrometry (LC-ESI-LTQ-Orbitrap-MS) combined with de novo software and UniProt of MaxQuant software. Four peptides were identified from A_1a3c–p, including YGCC (444.1137 Da) and DSSCSG (554.1642 Da) identified by de novo software and NNAEYYK (900.3978 Da) and PAGNVR (612.3344 Da) identified by UniProt of MaxQuant software. The molecular weights and amino acid sequences of four peptides were in accordance with the features of antioxidant peptides. The results indicated that different peptides were identified by different data analysis software according to spectrometry mass data. Considering the complexity of LC-ESI-LTQ-Orbitrap-MS, it was necessary to use the different methods to identify the key peptides from protein hydrolysates.     

Kinetics of the Inhibition of Renin and Angiotensin I Converting Enzyme by Polar and Non-polar Polyphenolic Extracts of <Emphasis Type="Italic">Vernonia Amygdalina</Emphasis> and <Emphasis Type="Italic">Gongronema Latifolium</Emphasis> Leaves

The aim of this study was to determine the in vitro modulation of the renin-angiotensin system by polyphenolic extracts and fractions of two green leafy vegetables, Vernonia amygdalina (VA) and Gongronema latifolium (GL), that are used for food and medicinal purposes. An 80% acetone extract of each leaf was fractionated on silicic acid-packed column to give two main fractions: acetone eluate (flow-through) and ethanol eluate (column-bound), that consist mostly of chlorophyllic and non-chlorophyllic fractions, respectively. Column fractionation resulted in polyphenolic fractions that displayed higher potency against angiotensin converting enzyme (ACE) and renin than the crude acetone extracts; generally, the chlorophyllic fraction was more active than the non-chlorophyllic fraction. ACE-inhibitory activity was significantly higher (p < 0.05) for the chlorophyllic fraction of VA than GL, with IC₅₀ values of 0.207 and 0.413 mg/ml, respectively. Similarly, the chlorophyllic fraction of VA had significantly higher (p < 0.05) renin inhibition than GL, with IC₅₀ values of 0.172 and 0.513 mg/ml, respectively. Kinetics studies showed that the chlorophyllic fractions of VA and GL exhibited mostly mixed-type ACE and renin inhibitions. We concluded that the hydrophobic nature of the chlorophyllic fraction may have contributed to the increased interaction with enzyme protein and inhibition of activities of ACE and renin.