Green Synthesis of Gold Nanoparticles in Pomegranate Seed Oil Stabilized Using Laser Ablation

In this paper, gold nanoparticles were synthesized in pomegranate seed oil using a laser ablation. The nano fluids were characterized using UV–Vis spectroscopy, transmission electron microscopy and Fourier transform infrared spectroscopy. The nanoparticles were formed inside the pomegranate seed oil in spherical form, and the particle size ranged from 12 to 4 nm, and the gold nanoparticles were capped with COO^? of carboxylic acid groups of fatty acid in pomegranate seed oil.     

Extraction of pomegranate seed oil using subcritical propane and supercritical carbon dioxide

Extraction of oil from pomegranate seeds as a waste product of the juice industry using supercritical carbon dioxide and subcritical propane was studied in this work. The influence of the main operating conditions of extraction, namely, the temperature and pressure of extraction on the oil extraction yield and the correspondent fatty acid profile were analyzed and reported here. Soxhlet extraction with n-hexane was done for comparison with supercritical extraction resulting in a maximum yield of oil of 22.31 wt %. Supercritical carbon dioxide and subcritical propane extracted up to 58.53% (corresponding to maximum yield of 13.06 wt %) and 76.73% (corresponding to maximum yield of 17.12 wt %) of the total amount of pomegranate seed oil as measured using Soxhlet extraction, respectively. Results indicated that the subcritical propane is a suitable and selective solvent for the extraction of the pomegranate seed oil in function of smaller times and pressures employed compared to carbon dioxide extraction. The fatty acid composition of the extracted oil showed the presence of fatty acids of C₁₆, C₁₈, C₂₀, C₂₂, and C₂₄ carbon chains. Punicic (C_18.3) was the major fatty acids and comprise up to 70% of the total fatty acid content of the extracted oil among all samples. Finally, Lack??s plug flow model as developed by Sovova was applied to both extraction systems and a good agreement with the experimental results was obtained.     

A Kinetic Approach to the Oxidation of Linseed Oil as Influenced by Fruit Peel and Seeds of Pomegranate

The objective of this research is to analyze the kinetic parameters of linseed oil by treating it with pomegranate peel and seeds at 353, 368, and 383 K using Rancimat. There are no significant differences (p < 0.05) between the oxidative stability indices of samples containing pomegranate peel and pomegranate seeds. In addition, the indices pertaining to the oxidative stability of linseed oil increase in value as the concentrations of pomegranate peel and seeds increase. Apart from the pomegranate peel at 0.1% and the quercetin, all other antioxidants are able to reduce the severity of temperature‐related parameters (i.e., temperature coefficient and Q₁₀ values). In addition, these antioxidants are able to form an activated complex with lower levels of thermal energy (by reducing activation energy and enthalpy) but of more structured configuration (by reducing the frequency factor and entropy). A high correlation is found between the Gibbs free‐energy of activation and the oxidative stability index of samples. The most substantial increase in the Gibbs free‐energy of activation occurs by TBHQ, followed by gallic acid, quercetin, and pomegranate peel at 1%. Practical Applications: Linseed oil is characterized by its high amounts of essential polyunsaturated fatty acids (PUFAs). The high PUFA content of linseed oil contributes to its rapid oxidation. Pomegranate peel and seeds are valuable sources of natural antioxidants. Investigating the effect of pomegranate peel and seeds in reducing the dependence of linseed oil oxidation on temperature provides a range of valuable kinetic parameters. Nevertheless, this subject has received little attention so far. There are few published data regarding the effect of natural antioxidants on lipid oxidation by this approach. Accordingly, this study is designed to investigate the oxidation kinetics and mechanisms of linseed oil as manipulated by fruit peel and seeds of pomegranate. The results show that the fruit peel and seeds of pomegranate can improve the oxidative stability of linseed oil and reduce the severity of effects caused by undesirable temperatures that may increase the oxidation rate of linseed oil.     

Comparative analysis of PVA hydrogels incorporating two natural antimicrobials: Punica granatum and Arnica montana tinctures

Poly(vinyl alcohol) (PVA) hydrogels are biocompatible materials that retain relatively high amounts of aqueous fluids in their network. The incorporation of antimicrobial natural products, such as pomegranate and arnica, is a strategy for coupling their bactericidal effects with the favorable properties of the gels for wound healing treatment. The goal of this work was to investigate the microstructural, mechanical, and release properties of PVA–pomegranate and PVA‐arnica samples. Physical interactions were observed by Fourier transform infrared spectra in PVA–pomegranate samples. The addition of pomegranate to PVA led to high T_g, probably related to the physical interactions, and low T_m and X_c, possibly due to the pomegranate location between PVA chains. Increased levels of arnica were associated with a slight decrease in samples' X_c, probably due to a physical barrier imposed by arnica to the interaction of PVA chains. PVA samples presented higher swelling than the samples containing natural products. High amounts of pomegranate in the samples led to high weight loss and high phenols and flavonoids delivery. Arnica release seemed to be more difficult than the pomegranate release. All samples presented relative high mechanical properties. In addition, pomegranate samples present higher potential to deliver active compounds than arnica samples. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45392.     

Evaluation of plant-based oils for production of offset printing ink

The oils and solvents are the main components of the printing ink, and the chemical composition of the ink could be harmful or toxic to human health and the environment. Therefore, there is an increasing demand to develop inks containing green, biobased, sustainable, and renewable raw materials instead of petrochemical substances. In this study, flaxseed oil (FO), pomegranate seed oil, plum kernel oil (PKO), and grape seed oil (GSO) were selected to produce offset printing inks. Pinus pinaster (P. pinaster) resin was also used in the formulation of inks to examine the effects of natural resin on ink together with vegetable oils. The phenolic content was analyzed for the resin and oils to figure out their potential antioxidant and bioactive characteristics. Optical and rheological tests were applied to evaluate the printability of the inks. L*a*b*, ΔE, density, and gloss tests were performed for optical evaluation. The viscosity, tack, and rub resistance tests were applied to perform rheological analysis. The biobased, environmental friendly, and self-drying (cold set) offset printing inks were obtained using natural pine resin and three different plant-based oils FO, PKO, and GSO. The printability analysis of the inks figured out the potential usage of plant-based oils in the offset printing ink formulation.     

Carbon dioxide extraction of canola seed: Oil solubility and effect of seed treatment

The extraction of oil from fixed beds of canola seed (Brassica napus) was studied using carbon dioxide at temperatures and pressures ranging from 25 to 90°C and 10 to 36 MPa, respectively. The oil solubility in CO₂ was found to be strongly dependent on CO₂ pressure and weakly dependent on the system temperature. The highest observed oil solubility was 11 mg/g CO₂ and occurred at 36 MPa and 55°C. The manner in which different methods of seed pretreatment (flaking, cooking, pressure rupturing, chopping and crushing) affected the extraction process also was studied. The total amount of oil recovered from the seeds by CO₂ extraction was found to be strongly dependent on the pretreatment. No measurable quantity of oil chould be recovered from whole, intact seeds. The amount of oil extractable from flaked and cooked seeds was comparable to that recoverable by conventional hexane extraction.     

Soybean Seed Quality Characteristics in Response to Indigenous Bradyrhizobium Inoculation and N Fertilization in Kashmir–Pakistan

The objective of this study was to quantify changes in soybean seed quality characteristics in response to indigenous Bradyrhizobium inoculation and N fertilization applied under field conditions during the years 2009 and 2010. Seven indigenous Bradyrhizobium isolates were isolated from the different locations under the foothills of great Himalayas Rawalakot Kashmir, Pakistan. The field isolates were compared to a reference strain (exotic) TAL102, three N fertilizer rates and to an un‐inoculated control. The seed oil content, fatty acid composition, seed N, P and K concentration and seed N, P and K uptake were quantified. Bradyrhizobium inoculation and N fertilization significantly increased oil content compared to the un‐inoculated control. The seed oil content varied between 16.2 and 21.5 %, highest in the seeds treated with indigenous Bradyrhizobium strains NR₂₂, NR₂₅ and NR₃₅, and mainly composed of linoleic acid (47 %), and oleic acid (24 %). Inoculation and N fertilization both decreased the saturated fatty acids (palmitic and stearic) and increased unsaturated fatty acids (linoleic acid and oleic acid). The mineral nutrient content of N, P, and K and their accumulation in seed increased by 2–3‐fold compared to the corresponding control. Indigenous Bradyrhizobium strains were able to generate the highest values for seed oil content (NR₂₂, NR₂₅, and NR₃₅), unsaturated fatty acids, i.e. linoleic acid and oleic acid (NR₂₅, and NR₃₅) and N, P and K uptake (NR₂₀, and NR₂₂). There were noticeable differences among the treatments in terms of essential fatty acid, oil, and mineral nutrient content. The study demonstrates the potential benefits of using indigenous Bradyrhizobium strains in order to achieve high quality soybean seeds that can be used as a balanced health product.     

Determination of conjugated linolenic acid content of selected oil seeds grown in Turkey

Seeds originating from some Turkish sources were analyzed with respect to their characteristics and FA compositions. These seeds represented pomegranate (Punica granatum L.), bitter grourd (Momordica charantia L.), pot marigold (Calendula officinalis L.), catalpa (Catalpa bignonoides), bourdaine (Rhamnus frangula L.), Oregon grape (Mahonia aquifolium), sarsaparilla (Smilax aspera), mahaleb (Prunus mahaleb L.), black-thorn (Prunus spinosa L.), cherry laurel (Prunus laurocerasus L.), and firethorn (Pyracantha coccinea). Bitter gourd and bourdaine seeds contain more than 20% oil. Catalpa, bourdaine, Oregon grape, blackthorn, and cherry laurel seed oil contents ranged from 15 to 20%. In the seeds from plants belonging to the Rosacea family, oil content ranged from 4.5 to 18.5%. Among the seed oils analyzed, pot marigold had one of the lowest oil contents (5.9%). Pomegranate contained the highest amount of total conjugated linolenic acid (CLNA) (86.0%). Seed oils of bitter grourd, pot marigold, mahaleb, and catalpa were rich in CLNA: 60.0, 29.5, 27.6, and 27.5%, respectively. Bourdaine, Oregon grape, and sarsaparilla seeds contained low amounts of CLNA. On the other hand, mahaleb, bourdaine, catalpa, Oregon grape, sarsaparilla, cherry laurel, blackthorn, and firethorn seed oils are basically oleic and linoleic acid-rich oils and therefore have little drying ability (semidrying oil). The results show a potential for the use of endogenous Turkish seeds as a source of CLNA.     

Relation between diacylglycerol acyltransferase activity and oil concentration in soybean

Diacylglycerol acyltransferase (EC 2.3.1.20; DGAT) catalyzes synthesis of triacylglycerol from acyl-CoA and diacylglycerol. Activity of this enzyme and developmental changes in oil accumulation were estimated at various stages of seed growth in soybean germplasm with phenotypic differences in oil content. Oil deposition in seed of these genotypes followed a sigmoid pattern that was modeled to predict incremental rates of oil accumulation during seed development. A strong positive correlation was found between the estimated peak rate of oil deposition (near the mid-term of seed development) and oil concentration in mature seed. At saturating substrate levels, DGAT activity measured near the peak rate of oil deposition also was correlated positively with oil phenotype. In the latter stages of seed development, a positive correlation between estimates of enzyme activity at or below the apparent K _m for diolein and comparable oil accumulation rates was attributed to reduced synthesis of substrates and/or potential change in affinity for substrate as suggested by an increase in apparent K _m for diolein in older seed. These data indicated that DGAT activity may be a rate-limiting step in triacylglycerol synthesis. However, it is difficult to accept the idea of a single rate-limiting step at the end of a complex metabolic pathway. Because oil is a quantitatively inherited trait, several genes determine genotypic differences in oil content among soybeans. Hence, DGAT activity may be an indicator of coordinated genetic expression of gene-products in the entire glycerolipid synthetic pathway for a given genotype. In any case, results of this investigation demonstrated that genotypic differences in DGAT activity contributed to expression of genetic variation in oil content among soybean gemplasm.     

Oxygen plasma pretreatment improves dyeing and antimicrobial properties of wool fabric dyed with natural extract from pomegranate peel

This paper presents the application of conventional potassium aluminium sulphate wool mordanting and a biochemical method with silver nitrate (as antimicrobial agent and mordant)_, alone and in combination with oxygen plasma, as part of comprehensive research into pretreatment processes for wool dyeing with natural extract from pomegranate peel (Punica granatum L.). Pretreatment with oxygen plasma significantly improved the hydrophilicity and tensile strength of all tested samples and showed that oxygen plasma can improve K/S, washing fastness, and even replace certain mordants in wool dyeing with natural pomegranate dye. All dyed samples exhibited good antibacterial activity against Staphylococcus aureus, which can be contributed to the phenol content in pomegranate dye. Only after 28 days of intensive ageing in natural weathering conditions did K/S and antibacterial activity against Klebsiella pneumoniae slightly decrease in dyed samples pretreated with oxygen plasma.     

Supercritical fluid extraction of daphne (Laurus nobilis L.) seed oil

Laurus nobilis L., commonly known as daphne tree, is an evergreen that belongs to the Lauraceae family. Daphne trees produce grape-sized shiny purplish berries having three parts: flesh, skin, and an inner kernel (single seed). This study examines supercritical CO₂ (SC-CO₂) extraction of oil from daphne seeds. The oil yield of ground seeds varied from 14 to 28% depending on the method and particle size used for oil recovery. Yields were similar for both petroleum ether and SC-CO₂ extraction. The extraction yield decreased significantly with increasing particle size. The amount of extract collected increased exponentially with increasing SC-CO₂ pressure. The highest extraction yield was obtained at the highest temperature studied, 75°C. More than 45% of the oil was lauric acid. SC-CO₂ is a viable technique to obtain high-purity L. nobilis L. seed oil, which is a potential ingredient for the cosmetic industry.     

Evaluation of the Triglyceride Composition of Pomegranate Seed Oil by RP‐HPLC Followed by GC‐MS

Triglyceride composition and fatty acid profiles of pomegranate seed oil were evaluated by newly developed methods in reverse‐phase‐high performance liquid chromatography (RP‐HPLC) and gas chromatography (GC), respectively. Different compositions of the mobile phase (acetone and acetonitrile) and flow rates for the HPLC system were used to obtain better separation for accurate quantitative analysis. Triglycerides with conjugated fatty acids (CLnAs) were eluted in order of the polarity of their geometrical isomers (c, t, c < t, t, c < t, t, t). The dominant triglyceride was found to be PuPuPu (32.99 %) in pomegranate seed oil, followed by PuPuCa and PuCaCa containing punicic acid and catalpic acid with total triglyceridelevels of 27.72 and 10.11 %, respectively. For fatty acid composition analysis, triglyceride fractions were derivatized into their respective methylesters which were injected into gas chromatography‐mass spectrometry (GC‐MS) to identify and gas chromatography‐flame ionization detector (GC‐FID) to quantify the conjugated fatty acids of each fraction of triglycerides. Punicic acid was found to be dominant (76.57 %) followed by catalpic acid (6.47 %) and β‐eleotearic acid (1.45 %). Pomegranate seed contained greater amounts of conjugated linolenic acids. These results showed that the present study provides more information about the composition of the triglyceride and fatty acid profiles of pomegranate seed oil compared to the reported studies. Therefore, the developed methods in this study can be used for the identification of the triglyceride and fatty acid composition for pomegranate seed oils and some such specials edible oils including CLnA isomers.     

Determination of seed oil content and fatty acid composition in sunflower through the analysis of intact seeds, husked seeds, meal and oil by near-infrared reflectance spectroscopy

A methodological study was conducted to test the potential of near-infrared reflectance spectroscopy (NIRS) to estimate the oil content and fatty acid composition of sunflower seeds. A set of 387 intact-seed samples, each from a single plant, were scanned by NIRS, and 120 of them were selected and further scanned as husked seed, meal, and oil. All samples were analyzed for oil content (nuclear magnetic resonance) and fatty acid composition (gas chromatography), and calibration equations for oil content and individual fatty acids (C_16:0, C_16:1, C_18:0, C_18:1, and C_18:2) were developed for intact seed, husked seed, meal, and oil. For intact seed, the performance of the calibration equations was evaluated through both cross- and external validation, while cross-validation was used in the rest. The results showed that NIRS is a reliable and accurate technique to estimate these traits in sunflower oil (validation r ² ranged from 0.97 to 0.99), meal (r ² from 0.92 to 0.98), and husked seeds (r ² from 0.90 to 0.97). According to these results, there is no need to grind the seeds to scan the meal; similarly accurate results are obtained by analyzing husked seeds. The analysis of intact seeds was less accurate (r ² from 0.76 to 0.85), although it is reliable enough to use for pre-screening purposes to identify variants with significantly different fatty acid compositions from standard phenotypes. Screening of intact sunflower seeds by NIRS represents a rapid, simple, and cost-effective alternative that may be of great utility for users who need to analyze a large number of samples.     

Development of calibration equations to predict oil content and fatty acid composition in brassicaceae germplasm by near-infrared reflectance spectroscopy

The use of nondestructive analytical methods is critical for the evaluation of very small seed samples such as those from germplasm collections. The objective of this study was to evaluate the potential of near-infrared reflectance spectroscopy (NIRS) for the simultaneous analysis of seed oil content and concentration of major fatty acids in intact-seed samples of the family Brassicaceae. A total of 495 samples from 56 genera and 128 species were analyzed by NIRS. The fatty acid composition of the seed oil was determined in all the samples by gas-liquid chromatography (GLC). The total seed oil content was determined by solvent extraction in 129 samples from 22 genera. Calibration equations for oil content (n=97) and individual fatty acids (n=410) were developed and tested through external validation with the samples not included in the calibration sets. The calibration equations for oil content (r ²=0.97 in validation) and concentrations of C_18:1 (r ²=0.93), C_18:3 (r ²=0.95), and C_22:1 (r ²=0.94) showed very good performance and provided reliable estimations of these traits in the samples of the validation set. The calibration equations for C_16:0, C_18:0, and C_18:2 content were less reliable, with r ² from 0.67 to 0.73. There was practically no response of NIRS to differences in C_20:1 (r ²=0.31). These results demonstrated that the oil content and concentrations of C_18:1, C_18:3, and C_22:1 can be estimated reliably within the family Brassicaceae by using NIRS calibration equations integrating broad taxonomic variability.     

Supercritical Carbon Dioxide Extraction of <Emphasis Type="Italic">Microula sikkimensis</Emphasis> Seed Oil

The seed oil of Microula sikkimensis had been intensively studied due to its pharmacological actions. In the present study, seed oil of Microula sikkimensis was extracted using supercritical fluid extraction (SFE). Determinations of the extracts composition were performed by gas chromatography (GC). An orthogonal array design (OAD), OA₉ (3⁴), was employed for optimization of the supercritical fluid extraction of the compound with regard to the various parameters. Four factors, namely pressure (21.0–27.0 MPa), the dynamic extraction time, temperature, and CO₂ flow rate of the supercritical fluid, were studied and optimized by a three-level OAD. The effects of the parameters on the yield of seed oil were studied using analysis of variance (ANOVA). The results revealed that the pressure had a significant effect on the yield of seed oil (p < 0.05), while the other three factors, i.e., CO₂ flow rate, dynamic extraction time and temperature, were not identified as significant factors under the selected conditions based on ANOVA. The results show that the best values for the extraction condition of seed oil was pressure 24.0 MPa, extraction time 3 h, temperature 45 °C and a CO₂ flow rate 20 L/h in the 20-L vessel.     

<Emphasis Type="Italic">Geranium sanguineum</Emphasis> (Geraniaceae) seed oil: A new source of petroselinic and vernolic acid

The occurrent of petroselinic acid (18∶1Δ6cis) in seed oils was believed to be limited to the Umbelliferae or Apiaceae, and a few other members of the Umbelliflorae. A major occurrence of petroselinic acid outside the Umbelliflorae must therefore be regarded as highly unusual and surprising. The seed oil of Geranium sanguineum, a member of the family Geraniaceae, has now been found to contain petroselinic and vernolic acids as major FA in its seed oil TAG. These unusual FA have not been reported previously as constituents of Geraniaceae seed oils. The structure and composition of the seed oil FA from G. sanguineum were determined by combined use of chromatographic (TLC, capillary GLC) and spectroscopic (IR, GC-MS) techniques. The double-bond position in petroselinic acid was located unambiguously by the characteristic mass fragmentation of its dimethyldisulfide (DMDS) adduct. The epoxy FA was identified as vernolic acid by co-chromatography and by the mass fragments formed during GC-MS of the products of the epoxy ring-opening reaction with BF₃ in methanol.     

Thermal dehydrochlorination of poly(vinyl chloride) in the presence of Jatropha seed oil

Thermal degradation of poly(vinyl chloride) was studied in a nitrogen atmosphere in the presence of Jatropha seed oil, epoxidized Jatropha seed oil, and soaps (barium and cadmium) of Jatropha seed oil at various temperatures. The rate of dehydrochlorination measurement at 1% degradation, R_DH, and the time required for dehydrochlorination to attain 1% conversion were used to assess the effect of the additives on the susceptibility of the polymer to dehydrochlorination. It was found from the kinetic studies and the results from viscosity measurements on degraded polymer samples that the Jatropha seed oil derivatives suppressed the initial loss of HCl from the polymer and the extent of polymer chain scission accompanying the dehydrochlorination process. Thermal degradation studies of poly(vinyl chloride) in the presence of mixtures of barium and cadmium soaps of Jatropha seed oil were also carried out. It was found that soap mixtures containing less than 80 wt% cadmium soap exerted a deleterious (antagonistic) effect on the degradation of poly(vinyl chloride) while in the presence of soaps containing more than 80 wt% cadmium soap, considerably lower values of R_DH and higher values of t_DH were observed. The soap mixture containing 90 wt% cadmium soap was found to exhibit a remarkedly improved stabilizing effect on the dehydrochlorination of poly(vinyl chloride). © 1995 John Wiley & Sons, Inc.     

Laboratory antimicrobial activity of cinnamaldehyde and pomegranate‐based polycaprolactone films

Polycaprolactone (PCL) was incorporated separately with cinnamaldehyde (CNMA), pomegranate methanolic extract (PME), freeze dried arils of pomegranate (FDAP), and seed flour of pomegranate (SF) to form antimicrobial films to be used for active food packaging. PCL–CNMA films completely inactivated growth of the artificially inoculated Escherichia coli and Staphylococcus aureus, whether at 5% or 10% concentrations (wt/wt of polymer), at all studied temperatures (4, 20, and 37 °C). PCL–PME films (10% wt/wt of polymer) delayed the growth of E. coli and S. aureus for up to 7 and 6 days, respectively, at 37 °C. Two‐day delays in the growth of both bacteria were achieved with FDAP and SF films (10% wt/wt of polymer) at 20 °C. The release of CNMA was slower than the release of PME during film processing, as measured by Fourier transform infrared spectroscopy. Thus, CNMA and pomegranate‐based films perhaps should be planned further for use in controlled release food packaging. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017 , 134, 45347.     

Determination of solution viscosity characteristics of rubber seed oil based alkyds resins

Three medium oil alkyd samples of different oil length [(I) 45%, (II) 50%, and (III) 55%] were synthesized with rubber seed oil. Dilute solution viscosity measurements were carried out on the alkyd in acetone and in toluene. The parameters investigated include intrinsic viscosity [η], Huggins constant (k_H), and Mark‐Houwink Sakurada constants (κ and α). The [η] values for the alkyd samples were found to be larger in acetone than in toluene. The K_H values showed a regular order in acetone but not in toluene. The K_H values showed no regular order in their variation with the oil content of the alkyd samples and the solvent, but the values obtained are higher in acetone than in toluene. Correlation of molecular weight (M) with [η] was also examined. [η] was observed to increase with the increase in the molecular weight of the resins. The α values obtained are in reasonable agreement with the reported ranges of α values in good solvent. The characteristics examined suggest that acetone is a better solvent for rubber seed oil alkyd resin than toluene. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 101: 3073–3075, 2006     

Cytotoxic effect of conjugated trienoic fatty acids on mouse tumor and human monocytic leukemia cells

The cytotoxicity of fatty acids from seed oils containing conjugated linolenic acids (CLN) was studied. Fatty acids from pomegranate, tung, and catalpa were cytotoxic to human monocytic leukemia cells at concentrations exceeding 5 μM for pomegranate and tung and 10 μM for catalpa, but fatty acids from pot marigold oil had no effect at concentrations ranging up to 163 μM. The main conjugated fatty acids of pomegranate, tung, catalpa, and pot marigold were cis(c)9, trans(t)11, c13-CLN (71.7%), c9,t11,t13-CLN (70.1%), t9,t11,c13-CLN (31.3%), and t8,t10,c12-CLN (33.4%), respectively. Therefore, the cytotoxicities of fatty acids from pomegranate, tung, and catalpa were supposed to be due to 9,11,13-CLN isomers. To elucidate the cytotoxicity of these CLN, we separated each CLN isomer from the fatty acid mixtures by high-performance liquid chromatography and analyzed its cytotoxicity. The cytotoxicities of c9,t11,c13-CLN, c9,t11,t13-CLN, and t9,t11,c13-CLN were much stronger than that of t8,t10,c12-CLN. Therefore, the higher cytotoxicity of fatty acids from pomegranate, tung, and catalpa than those from pot marigold would be derived from the different activities of 9,11,13-CLN and 8,10,12-CLN. Since there was little difference in the cytotoxicities of c9,t11,c13-CLN, c9,t11,t13-CLN, and t9,t11,c13-CLN, it is suggested that the cis/trans configuration of 9,11,13-CLN isomers had little effect on their cytotoxic effects. The mechanism of the cytotoxicity of the four fatty acids above may involve lipid peroxidation, because the order of toxicity of the fatty acids was consistent with their susceptibility to peroxidation in aqueous phase. This was supported by the decrease in the cytotoxicity of the fatty acids by addition of butylated hydroxytoluene.