Antioxidant activity of oregano, parsley, and olive mill wastewaters in bulk oils and oil-in-water emulsions enriched in fish oil

The antioxidant activity of oregano, parsley, olive mill wastewaters (OMWW), Trolox, and ethylenediaminetetraacetic acid (EDTA) was evaluated in bulk oils and oil-in-water (o/w) emulsions enriched with 5% tuna oil by monitoring the formation of hydroperoxides, hexanal, and t-t-2,4-heptadienal in samples stored at 37 degrees C for 14 days. In bulk oil, the order of antioxidant activity was, in decreasing order (p < 0.05), OMWW > oregano > parsley > EDTA > Trolox. The antioxidant activity in o/w emulsion followed the same order except that EDTA was as efficient an antioxidant as OMWW. In addition, the total phenolic content, the radical scavenging properties, the reducing capacity, and the iron chelating activity of OMWW, parsley, and oregano extracts were determined by the Folin-Ciocalteau, oxygen radical absorbance capacity, ferric reducing antioxidant power, and iron(II) chelating activity assays, respectively. The antioxidant activity of OMWW, parsley, and oregano in food systems was related to their total phenolic content and radical scavenging capacity but not to their ability to chelate iron in vitro. OMWW was identified as a promising source of antioxidants to retard lipid oxidation in fish oil-enriched food products.     

Antioxidant behavior in bulk oil: limitations of polar paradox theory

The polar paradox theory that explains the efficacy of antioxidants as affected by their polarity and that of the medium involved was re-evaluated. For the first time, the effect of concentration on validity of the polar paradox theory was investigated using four pairs of polar and nonpolar representative antioxidants in bulk oil. A model on antioxidant behavior in response to their polarity is proposed.     

Antioxidant activity of capsaicinoid in canola oil

Interest in replacing synthetic antioxidants, namely, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), with natural antioxidants is increasing. The present study examined the antioxidant activity of capsaicinoid from chili pepper in heated canola oil. The oxidation was conducted at 60, 90, 120, and 180 °C by monitoring oxygen consumption and the decrease in linoleic acid and α-linolenic acid in canola oil. At 60 °C, capsaicinoid was more effective against oxidation of canola oil compared with BHT. At higher temperatures of 90, 120, and 180 °C, capsaicinoid possessed an antioxidant activity similar to or slightly weaker that that of BHT. It was found that capsaicinoid prevented canola oil from oxidation in a dose-dependent manner. To study the structure-antioxidant relationship, it was found that the trimethylsiloxy (TMS) derivatives of capsaicinoid did not exhibit any antioxidant activity, suggesting the hydroxyl moiety was the functional group responsible for the antioxidant activity of capsaicinoid. It was concluded that capsaicinoid had the potential to be further explored as a natural antioxidant in foods, particularly spicy foods.     

Antioxidant activity of cysteine, tryptophan, and methionine residues in continuous phase beta-lactoglobulin in oil-in-water emulsions

Proteins dispersed in the continuous phase of oil-in-water emulsions are capable of inhibiting lipid oxidation reactions. The antioxidant activity of these proteins is thought to encompass both free radical scavenging by amino acid residues and chelation of prooxidative transition metals; however, the precise mechanism by which this occurs remains unclear. In this study, the oxidative stability of cysteine, tryptophan, and methionine residues in continuous phase beta-lactoglobulin (beta-Lg) in a Brij-stabilized menhaden oil-in-water emulsion was determined. The presence of low concentrations of continuous phase beta-Lg (250 and 750 microg/mL) significantly inhibited lipid oxidation as determined by lipid hydroperoxides and thiobarbituric acid reactive substances analysis. It was observed that cysteine oxidized before tryptophan in beta-Lg, and both residues oxidized before lipid oxidation could be detected. No oxidation of the methionine residues of beta-Lg was observed despite its reported high oxidative susceptibility. It is conceivable that surface exposure of amino acid residues greatly affects their oxidation kinetics, which may explain why some residues are preferentially oxidized relative to others. Further elucidation of the mechanisms governing free radical scavenging of amino acids could lead to more effective applications of proteins as antioxidants within oil-in-water food emulsions.     

Antioxidant properties of Teaw (Cratoxylum formosum Dyer) extract in soybean oil and emulsions

The antioxidant activity of an extract from Teaw (Cratoxylum formosum Dyer) leaves was studied in soybean oil and soybean oil-in-water emulsions. Samples containing the extract or reference antioxidants including chlorogenic acid, which comprises 60% of the Teaw extract, were stored at 60 degrees C and analyzed periodically for peroxide value (PV) and thiobarbituric acid reactive substances (TBARS) to allow both hydroperoxides and hydroperoxide degradation products to be monitored. Chlorogenic acid and the Teaw extract were more effective than alpha-tocopherol in inhibiting lipid oxidation in bulk oil but were less effective in an oil-in-water emulsion in accordance with the polar paradox. The PV/TBARS ratio for oil samples containing chlorogenic acid was higher than for alpha-tocopherol and BHT because chlorogenic acid inhibits both hydroperoxide formation by radical scavenging and hydroperoxide decomposition by metal chelation. The importance of the metal-chelating activity in retarding hydroperoxide decomposition was confirmed by studying the decomposition of oil samples containing added ferric ions. The PV/TBARS ratio was higher for citric acid than for alpha-tocopherol in the presence of added ferric chloride, but the order was reversed in samples lacking ferric chloride. Samples containing added chlorogenic acid gave the highest PV/TBARS ratios both in the presence and absence of ferric ions. The PV/TBARS ratios for the samples containing antioxidants fell rapidly to lower values in a soybean oil-in-water emulsion than in the soybean oil. This was due to increased hydroperoxide decomposition in the emulsion at the same PV. The Teaw extract contained 12% oil-soluble components, which contributed to a slightly higher oil-water partition coefficient than that of chlorogenic acid. The antioxidant activity of the aqueous phase of the Teaw extract was reduced more than that of chlorogenic acid by partitioning of the oil-soluble components into oil, which showed that the less-polar components contributed to the antioxidant activity of the Teaw extract in aqueous media.     

Effects of natural phenolic compounds on the antioxidant activity of lactoferrin in liposomes and oil-in-water emulsions

The effect of natural phenolic compounds on the antioxidant and prooxidant activity of lactoferrin was studied in liposomes and oil-in-water emulsions containing iron. The antioxidants tested with lactoferrin were alpha-tocopherol, ferulic acid, coumaric acid, tyrosol, and natural phenolic extracts obtained from three different extra-virgin olive oils and olive mill wastewater. The natural extracts of olive oils and mill wastewaters were composed mainly of polyphenols and simple phenolics, respectively. Lipid oxidation at 30 degrees C was determined by the formation of hydroperoxides and fluorescent compounds resulting from oxidized lipid interactions. All phenolic compounds showed synergistic properties in reinforcing the antioxidant activity of lactoferrin in lipid systems containing iron. The highest synergistic effects were observed for the phenolic extracts rich in polyphenols of extra-virgin olive oils and lactoferrin. This synergistic effect was higher in liposomes than in emulsions.     

Antioxidant activity and partitioning of phenolic acids in bulk and emulsified methyl linoleate.

To evaluate the effect of colloidal parameters on the activity of natural antioxidants, the effect of selected phenolic acids on both bulk and emulsified methyl linoleate oxidation (in the dark at 40 degrees C) was examined. Oxidation was monitored by determining the formation of hydroperoxides; their isomer distribution and ketodiene (oxodiene) products were monitored by using high-performance liquid chromatography. This study showed the system- and concentration-dependent antioxidant activity of phenolic acids. The scavenging of alpha,alpha-diphenyl-beta-picrazylhydrazyl radicals reflected the antioxidant activity in a bulk oil system but not in an emulsion. Specific interactions of the antioxidant with other compounds, for example, the emulsifier, and intramolecular hydrogen bonds may play an important role in reducing the antioxidant activity. Furthermore, these interactions of antioxidants with emulsifier have a strong influence on the partitioning of antioxidants. Thus, the proportion of the antioxidant solubilized in the lipid phase and particularly in the interface did not necessarily reflect the efficiency of the antioxidant.     

Characterization of the antioxidant activity of sugars and polyhydric alcohols in fish oil emulsions

Polyols have been incorporated into fish oil emulsions as a means for the inhibition of lipid oxidation and suppression of fishy flavor. However, the role of sugars and polyhydric alcohols as antioxidants has not been clearly established. Selected polyols were evaluated for their performance as antioxidants and modifiers of oxidation pathways in a model system. Oil/water (O/W) emulsions were prepared with freshly steam-deodorized menhaden oil. A layer of emulsion in aluminum pans held at 5 degrees C was exposed to 2550 lx fluorescent lights for 24 h before peroxide values and volatile flavor compounds were analyzed by GC headspace entrainment procedure. Antioxidant activity was confirmed for fructose, sucrose, raffinose, sorbitol, or mannitol when incorporated at 16% of the aqueous phase into model fish oil-in-water emulsions. Peroxide values were suppressed 10-18% in treated samples compared to control samples. Viscosity data did not exclude possible contributions from a restricted oxygen diffusion mechanism in the antioxidant activity, but revealed that emulsion viscosity did not govern fish oil oxidation rates. Combining polyols with phenolic antioxidants (alpha-tocopherol, BHT, or TBHQ) frequently diminished the antioxidant activity compared to that for individual phenolic antioxidants, which was interpreted as indicating that the H-donating activity of phenolic antioxidants was hindered by the H-bonding activity of polyols. A viscosity-based inhibition of the retroaldol conversion of (E,Z)-2,6-nonadienal to (Z)-4-heptenal with a high fructose concentration (67%) was attributed to a restriction of molecular mobility of reactants, but the conversion was only slightly inhibited by the concentration of fructose (16%) used in experimental emulsions. The data supported a hypothesis that either or both free radical scavenging and transition state metal chelation activities were provided by polyols in fish oil emulsions. Also, polyols retarded the water-requiring retroaldol decomposition of (E,Z)-2,6-nonadienal to (Z)-4-heptenal in the model systems and the reaction may be involved in some suppression of fishy flavors in emulsions.     

The relationship between the physicochemical properties of antioxidants and their ability to inhibit lipid oxidation in bulk oil and oil-in-water emulsions

Chain-breaking antioxidants differ in their effectiveness at inhibiting lipid oxidation because of their chemical properties and physical location within a food. Our objective was how the physicochemical properties of four structurally related lipid-soluble antioxidants were related to their antioxidant activity. Antioxidants differed in the number of methyl (alpha-tocopherol and delta-tocopherol) or hydroxyl (butylated hydroxytoluene (BHT) and 4-hydroxymethyl-2,6-ditertiarybutylphenol) groups. Surface activity of the antioxidants was in the order of delta-tocopherol > alpha-tocopherol approximately 4-hydroxymethyl-2,6-ditertiarybutylphenol > BHT. Free-radical scavenging activity was similar between alpha-tocopherol and delta-tocopherol as well as BHT and 4-hydroxymethyl-2,6-ditertiarybutylphenol. In bulk menhaden oil, BHT was a more effective antioxidant than 4-hydroxymethyl-2,6-ditertiarybutylphenol while alpha-tocopherol was more effective than delta-tocopherol. In menhaden oil-in-water emulsions, BHT was a more effective antioxidant than 4-hydroxymethyl-2,6-ditertiarybutylphenol while delta-tocopherol was more effective than alpha-tocopherol. These results indicate that the surface activity and polarity of lipid-soluble antioxidants were not the only determinants of their antioxidant effectiveness in food lipids.     

Purified phenolics from hydrothermal treatments of biomass: ability to protect sunflower bulk oil and model food emulsions from oxidation

The phenolic fractions released during hydrothermal treatment of selected feedstocks (corn cobs, eucalypt wood chips, almond shells, chestnut burs, and white grape pomace) were selectively recovered by extraction with ethyl acetate and washed with ethanol/water solutions. The crude extracts were purified by a relatively simple adsorption technique using a commercial polymeric, nonionic resin. Utilization of 96% ethanol as eluting agent resulted in 47.0-72.6% phenolic desorption, yielding refined products containing 49-60% w/w phenolics (corresponding to 30-58% enrichment with respect to the crude extracts). The refined extracts produced from grape pomace and from chestnut burs were suitable for protecting bulk oil and oil-in-water and water-in-oil emulsions. A synergistic action with bovine serum albumin in the emulsions was observed.     

Antioxidant activity of a proanthocyanidin-rich extract from grape seed in whey protein isolate stabilized algae oil-in-water emulsions

Algae oil-in-water emulsions stabilized with 0.2% whey protein isolate (WPI) at pH 3.0 and 7.0 were chosen to evaluate antioxidant activity of a proanthocyanidin-rich extract from grape seed. In this emulsion system, (+)-catechin and ascorbic acid (620 microM) were found to be prooxidative at pH 3.0 and ineffective at pH 7.0. Grape seed extract was not able to effectively inhibit both lipid hydroperoxides and propanal formation when added to the emulsion at 124 microM. However, increasing the concentration of the grape seed extract to 620 microM resulted in inhibition of both lipid hydroperoxide and propanal formation at pH 3.0 and 7.0. None of the antioxidants tested had any effect on the physical stability of the WPI-stabilized emulsion. The superior antioxidant activity of the grape seed extract is likely due to the presence of oligomeric procyanidins which are better antioxidants compared to their monomeric counterparts.     

Antioxidant activity of olive pulp and olive oil phenolic compounds of the arbequina cultivar

The aim of this study was to characterize antioxidant activities of phenolic compounds that appear in olive pulp and olive oils using both radical scavenging and antioxidant activity tests. Antiradical and antioxidant activities of olive pulp and olive oil phenolic compounds were due mainly to the presence of a 3,4-dihydroxy moiety linked to an aromatic ring, and the effect depended on the polarity of the phenolic compound. Glucosides and more complex phenolics exhibited higher antioxidant activities toward oxidation of liposomes, whereas in bulk lipids aglycons were more potent antioxidants with the exception of oleuropein. Lignans acted as antioxidants only in liposomes, which could partly be due to their chelating activity, because liposome oxidation was initiated by cupric acetate. The antioxidant activity of virgin olive oil is principally due to the dialdehydic form of elenolic acid linked to hydroxytyrosol (3,4-DHPEA-EDA), a secoiridoid derivative (peak RT 36, structure unidentified), and luteolin.     

Antioxidant activity and bioactive compounds of tea seed (Camellia oleifera Abel.) oil

The oil of tea seed (Camellia oleifera Abel.) is used extensively in China as cooking oil. The objectives of this study were to investigate the antioxidant activity of tea seed oil and its active compounds. Of the five solvent extracts, methanol extract of tea seed oil exhibited the highest yield and the strongest antioxidant activity as determined by DPPH scavenging activity and Trolox equivalent antioxidant capacity (TEAC). Two peaks separated from the methanol extract by HPLC contributed the most significant antioxidant activity. These two peaks were further identified as sesamin and a novel compound: 2,5-bis-benzo[1,3]dioxol-5-yl-tetrahydro-furo [3,4-d][1,3]dioxine (named compound B) by UV absorption and characterized by MS, IR, 1H NMR, and 13C NMR techniques. Sesamin and compound B decreased H2O2-mediated formation of reactive oxygen species in red blood cells (RBCs), inhibited RBCs hemolysis induced by AAPH, and increased the lag time of conjugated dienes formation in human low-density lipoprotein. The results indicate that both compounds isolated from tea seed oil exhibit remarkable antioxidant activity. Apart from the traditional pharmacological effects of Camellia oleifera, the oil of tea seed may also act as a prophylactic agent to prevent free radical related diseases.     

Antioxidant activity of hydroxytyrosol acetate compared with that of other olive oil polyphenols

Hydroxytyrosol acetate was synthesized, and the antioxidant activity of this olive oil component was assessed in comparison with that of other olive oil components, namely hydroxytyrosol, oleuropein, 3,4-DHPEA-EA, and alpha-tocopherol in bulk oil and oil-in-water emulsions. The activity of the compounds was also assessed by scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. Hydroxytyrosol acetate had a weaker DPPH radical scavenging activity than hydroxytyrosol, oleuropein, or 3,4-DHPEA-EA but it had a radical scavenging activity similar to that of alpha-tocopherol. In oil, the antioxidant activity of hydroxytyrosol acetate was much higher than that of alpha-tocopherol or oleuropein, but in an emulsion 3,4-DHPEA-EA and alpha-tocopherol were more effective as antioxidants than hydroxytyrosol acetate. The antioxidant activity of hydroxytyrosol acetate was rather similar to that of hydroxytyrosol in oil and emulsions despite the difference in DPPH radical scavenging activity.     

Antioxidant activity of citrus limonoids, flavonoids, and coumarins

A variety of in vitro models such as beta-carotene-linoleic acid, 1,1-diphenyl-2-picryl hydrazyl (DPPH), superoxide, and hamster low-density lipoprotein (LDL) were used to measure the antioxidant activity of 11 citrus bioactive compounds. The compounds tested included two limonoids, limonin (Lim) and limonin 17-beta-D-glucopyranoside (LG); eight flavonoids, apigenin (Api), scutellarein (Scu), kaempferol (Kae), rutin trihydrate (Rut), neohesperidin (Neh), neoeriocitrin (Nee), naringenin (Ngn), and naringin(Ng); and a coumarin (bergapten). The above compounds were tested at concentration of 10 microM in all four methods. It was found that Lim, LG, and Ber inhibited <7%, whereas Scu, Kae, and Rut inhibited 51.3%, 47.0%, and 44.4%, respectively, using the beta-carotene-linoleate model system. Lim, LG, Rut, Scu, Nee, and Kae showed 0.5% 0.25%, 32.2%, 18.3%, 17.2%, and 12.2%, respectively, free radical scavenging activity using the DPPH method. In the superoxide model, Lim, LG, and Ber inhibited the production of superoxide radicals by 2.5-10%, while the flavonoids such as Rut, Scu, Nee, and Neh inhibited superoxide formation by 64.1%, 52.1%, 48.3%, and 37.7%, respectively. However, LG did not inhibit LDL oxidation in the hamster LDL model. But, Lim and Ber offered some protection against LDL oxidation, increasing lag time to 345 min (3-fold) and 160 min (33% increase), respectively, while both Rut and Nee increased lag time to 2800 min (23-fold). Scu and Kae increased lag time to 2140 min (18-fold) and 1879 min (15.7-fold), respectively. In general, it seems that flavonoids, which contain a chromanol ring system, had stronger antioxidant activity as compared to limonoids and bergapten, which lack the hydroxy groups. The present study confirmed that several structural features were linked to the strong antioxidant activity of flavonoids. This is the first report on the antioxidant activity of limonin, limonin glucoside, and neoeriocitrin.     

Density and microviscosity studies of palm oil/water emulsions

Electron paramagnetic resonance (EPR) and densitometry were used to measure the temperature- and rate-dependent formation of fat crystals in emulsion droplets in hardened palm kernel oil in water emulsions. The solid fat content in emulsions can be critical for the functionality of the emulsions in a wide variety of applications. Therefore, new and accessible methods are needed to monitor solid fat content in order to control the functional properties of these emulsions. EPR measurements showed that the microviscosity within the oil droplets could be measured as a function of temperature and that the storage temperature below the main fat melting point is crucial for an increase in the microviscosity, due to fat solidification. The microviscosity of the oil droplets could be an important parameter for defining functional performance (e.g., rheology and partial coalescence). Density measurements provided a simple and accurate method for monitoring changes in phase state of the oil. The phase behavior agreed well with the EPR results, demonstrating that accurate density measurements could prove to be a valuable tool for monitoring fat crystallization processes.     

Role of hydrocolloids in the creaming of oil in water emulsions

The influence of guar gum and xanthan gum on the creaming of 10% oil in water emulsions has been investigated. The presence of very low concentrations of the polysaccharides (typically < approximately 0.075%) was found to induce depletion flocculation of the emulsion droplets and increased the rate of creaming. However, at higher polysaccharide concentrations (typically > approximately 0.1%) the creaming rate was reduced due to the increased viscosity of the aqueous phase. For most systems a delay period was noted before creaming started, which was found to be dependent on the zero shear viscosity of the continuous phase and independent of polysaccharide type. The delay period increased significantly at zero shear viscosities approaching 1 Pa s. A mathematical model has been used to fit the creaming rate profiles and a simple exponential relationship obtained between delay time and polysaccharide concentration.     

Antioxidant evaluation and oxidative stability of structured lipids from extravirgin olive oil and conjugated linoleic acid

Structured lipid (SL) was synthesized from extravirgin olive oil (EVOO) and conjugated linoleic acid (CLA) via a lipase-catalyzed reaction. CLA provides a variety of health benefits, but it is not consumed in free fatty acid form. The synthesized SL olive oil contained 42.5 mol % CLA isomers, and the major isomers were cis-9,trans-11-CLA (16.9 mol %) and trans-10,cis-12-CLA (24.2 mol %). The antioxidant activity determined by the radical scavenging capacity with the 2,2-diphenyl-1-picrylhydrazyl radical was lower in SL olive oil than in EVOO. The oxidative stability was also lower in SL olive oil since it had a higher peroxide value, rho-anisidine value, and 2-thiobarbituric acid reactive substances values during 20 days of storage at 60 degrees C. This observation could be due to the reduction in the natural phenolic compounds (97%) and tocopherols (56%), and the incorporated CLA with two conjugated double bonds in the SL olive oil. The oxidative stability of SL olive oil was increased by added rosemary extracts at concentrations of 100, 200, and 300 ppm. The present study suggests that the SL olive oil may be a suitable way to incorporate or deliver CLA into human diets. However, the addition of a proper antioxidant would be required for improving its oxidative stability.     

Antioxidant activity of dietary oregano essential oil and alpha-tocopheryl acetate supplementation in long-term frozen stored turkey meat

The effects of dietary oregano essential oil and alpha-tocopheryl acetate supplementation on the oxidative stability of long-term frozen stored turkey meat were investigated. Thirty 12-week-old turkeys, randomly divided into five groups, were given a basal diet or a basal diet supplemented with 200 mg of alpha-tocopheryl acetate kg(-1), or 100 or 200 mg of oregano oil kg(-1), or 100 mg of oregano oil plus 100 mg of alpha-tocopheryl acetate kg(-1) for 4 weeks prior to slaughter. Lipid oxidation in breast and thigh meat was assessed after 1, 3, 6, and 9 months of frozen storage at -20 degrees C prior to or following 7 days of refrigerated storage at 4 degrees C. Results showed that oregano oil increased the oxidative stability of breast and thigh meat during the frozen storage. Dietary oregano oil at the inclusion level of 200 mg kg(-1) feed was significantly (p < 0.05) more effective in delaying lipid oxidation compared to the level of 100 mg kg(-1), but equivalent to dietary alpha-tocopheryl acetate supplementation at 200 mg kg(-1), which in turn was inferior to dietary supplementation of 100 mg kg(-1) oregano essential oil plus 100 mg kg(-1) alpha-tocopheryl acetate that was significantly (p < 0.05) superior to all other treatments. Thigh meat was more susceptible to oxidation than breast meat, although the former contained alpha-tocopherol at markedly higher levels. Mean alpha-tocopherol levels in breast and thigh meat from all treatments decreased during the frozen storage, the decrease being sharper between 1 and 3 months of frozen storage for breast and between 3 and 6 months for thigh meat. Oregano oil supplementation increased (p < 0.05) the retention of alpha-tocopherol in meat, the increase being positively correlated with the supplementation level. However, the retention of alpha-tocopherol in meat could only partly elucidate the antioxidant activity exhibited by dietary oregano oil supplementation.     

Antioxidant activity, cytotoxicity, and DNA information of Glossogyne tenuifolia

This study investigates the antioxidant activity and cytotoxicity of Glossogyne tenuifolia extract on various cancer cell lines. The 5.8s DNA of G. tenuifolia was isolated, and the species of this plant was confirmed by NCBI's DNA database. G. tenuifolia was then extracted with ethanol and separated into several fractions using the partition procedure with water, n-butanol, and ethyl acetate (EA). Among these, the EA fraction most significantly affected the activity of DPPH(*) and superoxide anion scavenging. Additionally, only the EA fraction exhibited cytotoxicity on breast cancer cells (MCF-7 and MDA-MB-231) and liver cancer cells (Hep G2 and Hep 3B). Next, the EA fraction was further separated by column chromatography, and 15 fractions were obtained. Three effective components were isolated and identified separately from the active fractions: oleanolic acid (OA) from fraction 6, luteolin from fractions 8-10, and luteolin-7-glucoside from fraction 12. The test of these three compounds on scavenging activity of DPPH(*) and superoxide anion indicates that luteolin had the highest antioxidant activity, whereas the effect of OA was negligible. Additionally, a synergistic effect between luteolin and luteolin-7-glucoside was observed. Kick-out experiments showed that the activities were vanished or decreased. Especially on MDA-MB-231 and MCF-7 cells, the cytotoxicity completely disappeared when luteolin was eliminated from fractions 8-10. These findings demonstrate that luteolin plays a crucial role in the inhibition of the growth of hepatoma cancer cell lines. Fraction 3, which did not contain luteolin, luteolin-7-glucoside, and oleanolic acid, had cytotoxicity on MDA-MB-231, MCF-7, Hep G2, Hep 3B, and A549, which implies that this fraction contained some other effective ingredients and requires further study. The investigation is currently underway in our laboratory.