Influence of fatty acids on wine foaming

The influence of fatty acids (free and bound as ethyl esters) on wine foaming was studied in different white wines and the corresponding sparkling wines. Moreover, from three of these wines the foam formed by CO(2) injection was separated, and two fractions were then obtained: foam wine (FW) and remainder wine (RW). In these fractions and the sparkling wines produced from them, foam properties and fatty acids were also determined. The free fatty acids C8, C10, and C12 were negatively correlated with foamability (HM), whereas the ethyl esters of hexanoic, octanoic, and decanoic acids were positively related to HM. The value of HM was directly proportional to the ratio of esterified to unesterified fatty acids. This was confirmed by the changes that occur in the esterification ratio during the second fermentation and aging. No influence was observed on either the Bikerman coefficient or the stability time of foam.     

Differential characteristics of fatty acids in cheese from milk of various animal species by capillary gas chromatography

The kind of milk used in the manufacture of cheese has been identified by analysis of the fatty acids. The milk fat is extracted from the cheese and saponified. The methyl esters of the fatty acids are prepared and determined by capillary column gas chromatography. Seven major fatty acids are separated and quantitated, namely, C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, and C18:1. Many of the 21 simple ratios that can be formed from these 7 quantities are characteristic of the type of milk from which the fatty acids were obtained. The method allows the identification of cheese prepared with the milk of cows, buffalo, sheep, or goats. Substitution or adulteration of milk can also be detected.     

A quantitative method for the determination of cyclopropenoid fatty acids in cottonseed, cottonseed meal, and cottonseed oil (Gossypium hirsutum) by high-performance liquid chromatography

Cyclopropenoid fatty acids (CPFAs), found in cottonseed, have been shown to have detrimental health effects to susceptible livestock. Previous quantitative analytical methods for the determination of CPFAs expressed these acids in terms of their relative abundance with respect to other fatty acids in the oil, necessitating the concurrent analysis of other fatty acids. The proposed analytical method describes the quantitation of three relevant CPFAs for cotton (malvalic acid, sterculic acid, and dihydrosterculic acid) in cottonseed in micrograms per gram fresh weight of sample. The method involves extraction of the oil, saponification, and derivatization of the free fatty acids with 2-bromoacetophenone to give the phenacyl esters. These esters are then separated by dual-column reverse-phase high-performance liquid chromatography and quantitated via external standards. This is the first method to include external calibration standards for CPFAs and, as such, is capable of direct quantification with no further data conversion required. CPFA data generated from the analysis of cottonseed, cottonseed meal, and cottonseed oil produced in the United States in 2002 are presented.     

Screening of the entire USDA castor germplasm collection for oil content and fatty acid composition for optimum biodiesel production

Castor has tremendous potential as a feedstock for biodiesel production. The oil content and fatty acid composition in castor seed are important factors determining the price for production and affecting the key fuel properties of biodiesel. There are 1033 available castor accessions collected or donated from 48 countries worldwide in the USDA germplasm collection. The entire castor collection was screened for oil content and fatty acid composition by nuclear magnetic resonance (NMR) and gas chromatography (GC), respectively. Castor seeds on the average contain 48.2% oil with significant variability ranging from 37.2 to 60.6%. Methyl esters were prepared from castor seed by alkaline transmethylation. GC analysis of methyl esters confirmed that castor oil was composed primarily of eight fatty acids: 1.48% palmitic (C16:0), 1.58% stearic (C18:0), 4.41% oleic (C18:1), 6.42% linoleic (C18:2), 0.68% linolenic (C18:3), 0.45% gadoleic (C20:1), 84.51% ricinoleic (C18:1-1OH), and 0.47% dihydroxystearic (C18:0-2OH) acids. Significant variability in fatty acid composition was detected among castor accessions. Ricinoleic acid (RA) was positively correlated with dihydroxystearic acid (DHSA) but highly negatively correlated with the five other fatty acids except linolenic acid. The results for oil content and fatty acid composition obtained from this study will be useful for end-users to explore castor germplasm for biodiesel production.     

脂肪酸甲酯生物柴油改善低硫柴油的润滑性能

生物柴油可作为改善低硫柴油润滑性能的天然添加剂。该文将豆蔻酸甲酯(C14:0)、棕榈酸甲酯(C16:0)、硬脂酸甲酯(C18:0)、油酸甲酯(C18:1)、亚油酸甲酯(C18:2)、亚麻酸甲酯(C18:3)、蓖麻醇酸甲酯(C18:1 OH)及蓖麻油甲酯和餐饮废油甲酯按照0.5%、1.0%、1.5%和3.0%的体积分数添加到低硫柴油中,在高频往复试验机(high-frequency reciprocating rig,HFRR)上进行润滑性能测试,探究脂肪酸甲酯的碳链长度、不饱和度及含羟基等结构特征对润滑性能的影响。结果表明,长碳链脂肪酸甲酯一般比短链润滑效果好;碳链长度为十八的脂肪酸酯中,不饱和程度即碳碳双键数目越高则润滑性能越好;而在相同碳链长度和不饱和度条件下,含羟基的蓖麻醇酸甲酯的润滑改善效果优于油酸甲酯。由多种脂肪酸酯构成的混合物生物柴油的润滑性能要优于某单一的纯脂肪酸甲酯。在低硫柴油中,当某饱和脂肪酸甲酯的体积分数比例达3.0%时,或不饱和酯的体积分数达到1.5%时,或生物柴油的体积分数达1.0%时,可使低硫柴油的润滑性能指标满足相关标准。研究脂肪酸甲酯的各种结构特征对其润滑性能的影响及作用机制,有助于筛选合适的生物柴油组分及其添加浓度作为低硫柴油的润滑添加剂。     

Strecker-type degradation of phenylalanine by methyl 9,10-epoxy-13-oxo-11-octadecenoate and methyl 12,13-epoxy-9-oxo-11-octadecenoate

The reaction of methyl 9,10-epoxy-13-oxo-11(E)-octadecenoate, methyl 12,13-epoxy-9-oxo-11(E)-octadecenoate, 4,5(E)-epoxy-2(E)-heptenal, and 4,5(E)-epoxy-2(E)-decenal with phenylalanine in acetonitrile-water (2:1, 1:1, and 1:2) at 80 degrees C and at different pHs and carbonyl compound/amino acid ratios was investigated both to determine if epoxyoxoene fatty esters were able to produce the Strecker-type degradation of the amino acid and to study the relative ability of oxidized long-chain fatty esters and short chain aldehydes with identical functional systems to degrade amino acids. The studied epoxyoxoene fatty esters degraded phenylalanine to phenylacetaldehyde. The mechanism of the reaction was analogous to that described for epoxyalkenals and is suggested to be produced through the corresponding imine, which is then decarboxylated and hydrolyzed. This reaction also produced a conjugated hydroxylamine, which was the origin of the long-chain pyridine-containing fatty ester isolated in the reaction and characterized as methyl 8-(6-pentylpyridin-2-yl)octanoate. Epoxyoxoene fatty esters and epoxyalkenals exhibited a similar reactivity for producing phenylacetaldehyde, therefore suggesting that nonvolatile lipid oxidation products, which are produced to a greater extent than volatile products, should be considered for determining the overall contribution of lipids to Strecker degradation of amino acids produced during nonenzymatic browning. In addition, the obtained data confirm that, analogously to carbohydrates, lipid oxidation products are also able to produce the Strecker degradation of amino acids.     

Gas chromatographic analysis of total fatty acids in cider

This paper reports the composition of total fatty acids in an apple beverage, cider. Fatty acids are present in the free or esterified form and contribute to both the flavor and foam properties of cider. Fatty acids were separated and identified as methyl esters by GC-MS, and 12 of these were subsequently determined by GC-FID. The major fatty acids found in cider were caproic, caprylic, capric, and palmitic acid, the saturated acids predominating over the unsaturated ones. The proposed method was applied to 59 ciders from three consecutive harvests (1996, 1997, and 1998), which were made by 19 cider-makers from the region of Asturias (Spain). Linear discriminant analysis of fatty acids in these samples allowed selection of palmitoleic, pentadecanoic, linoleic, myristic, and linolenic acid as the most predictive variables to differentiate ciders made from apples grown in the Asturias region (1997 harvest) and ciders made from apples grown outside this region (1996 and 1998 harvests).     

Isolation,structural elucidation,and inhibitory effects of terpenoid and lipid constituents from sunflower pollen on Epstein-Barr virus early antigen induced by tumor promoter,TPA

Eight fatty acid esters of triterpene alcohols (1-8), four free triterpene alcohols (9, 12, 17, and 18), four diterpene acids (19-22), two tocopherol-related compounds (23 and 24), four estolides (25-28), three syn-alkane-4,6-diols (29-31), one 1,3-dioxoalkanoic acid (32), and one aliphatic ketone (33), along with the mixture of free fatty acids, were isolated from the diethyl ether extract of the pollen grains of sunflower (Helianthus annuus). Among these compounds, 14 (2-8, 12, 23, 25-28, and 33) were new naturally occurring compounds, and their structures were determined on the basis of spectroscopic methods. Twenty-four terpenoids and lipids (1-4, 6-9, 12, and 19-33) and six free triterpene triols (10, 11, and 13-16), derived from their fatty acid esters (2, 3, and 5-8) by alkaline hydrolysis, were evaluated with respect to their inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), in Raji cells, which is known to be a primary screening test for antitumor promoters. Among the 30 compounds tested, 21 compounds possessing a di- or a polycyclic ring system in the molecule (1-4, 6-16, and 19-24) showed potent inhibitory effects on EBV-EA induction (91-100% inhibition at 1 x 10(3) mol ratio/TPA).     

Authentication of vegetable oils by bulk and molecular carbon isotope analyses with emphasis on olive oil and pumpkin seed oil

The authenticity of vegetable oils consumed in Slovenia and Croatia was investigated by carbon isotope analysis of the individual fatty acids by the use of gas chromatography-combustion-isotope ratio mass spectrometry (GC/C/IRMS), and through carbon isotope analysis of the bulk oil. The fatty acids from samples of olive, pumpkin, sunflower, maize, rape, soybean, and sesame oils were separated by alkaline hydrolysis and derivatized to methyl esters for chemical characterization by capillary gas chromatography/mass spectrometry (GC/MS) prior to isotopic analysis. Enrichment in heavy carbon isotope ((13)C) of the bulk oil and of the individual fatty acids are related to (1) a thermally induced degradation during processing (deodorization, steam washing, or bleaching), (2) hydrolytic rancidity (lipolysis) and oxidative rancidity of the vegetable oils during storage, and (3) the potential blend with refined oil or other vegetable oils. The impurity or admixture of different oils may be assessed from the delta(13)C(16:0) vs. delta(13)C(18:1) covariations. The fatty acid compositions of Slovenian and Croatian olive oils are compared with those from the most important Mediterranean producer countries (Spain, Italy, Greece, and France).     

Determination of cis, cis-methylene interrupted polyunsaturated fatty acids in fats and oils by capillary gas chromatography

A capillary gas chromatographic (CGC) method is described for the determination of cis,cis-methylene interrupted polyunsaturated fatty acids (cis-PUFA) in fats and oils. The sample is saponified and the liberated fatty acids are esterified to the corresponding methyl esters. The latter are analyzed by CGC using a 60 M. SP2340 capillary column. Area percent values for 9,12-cis,cis-C18:2 and 9,12,15-cis,cis,cis-C18:3 fatty acid methyl esters are summed to give the total cis-PUFA content. Gas chromatographic results agreed well with those obtained by an enzymatic lipoxygenase method at the 31-48% cis-PUFA levels with a correlation coefficient of 0.98. The method has a precision (relative standard deviation) of 0.33% at a 44.4% cis-PUFA level in margarine oil.     

Mass spectrometric identification and gas-liquid chromatographic determination of 2-chloroethyl esters of fatty acids in spices and foods.

The 2-chloroethyl esters of 5 fatty acids have been identified in spice and food samples by gas-liquid chromatography-mass spectrometry (GLC/MS). Twenty-four spice samples were analyzed for the 2-chloroethyl esters of fatty acids by AOAC official multiple residues pesticide procedure using GLC with microcoulometric detection. The esters of capric, lauric, myristic, palmitic, and linoleic acids have been identified at levels up to 1400 ppm. 2-Chloroethyl linoleate was the most abundant ester in all samples. Several foods analyzed by the same procedures showed levels of 2-chloroethyl linoleate as high as 35 ppm. Recoveries from fortified samples ranged from 84 to 98% for the various esters. A method using an acid-catalyzed esterification reaction was developed to rapidly determine the fatty acid content of these spices. GLC analysis with microcoulometric detection was used. Recoveries from fortified samples ranged from 92 to 110%. After 2 spice samples found to be free of 2-chloroethyl esters were fumigated with ethylene oxide, the level of 2-chloroethyl linoleate reached 77 ppm. All levels of 2-chloroethyl esters were confirmed by GLC/MS.     

Application of ethyl esters and d3-methyl esters as internal standards for the gas chromatographic quantification of transesterified fatty acid methyl esters in food

Ethyl esters (FAEE) and trideuterium-labeled methyl esters (d3-FAME) of fatty acids were prepared and investigated regarding their suitability as internal standards (IS) for the determination of fatty acids as methyl esters (FAME). On CP-Sil 88, ethyl esters of odd-numbered fatty acids eluted approximately 0.5 min after the respective FAME, and only coelutions with minor FAME were observed. Depending on the problem, one or even many FAEE can be added as IS for the quantification of FAME by both GC-FID and GC-MS. By contrast, d3-FAME coeluted with FAME on the polar GC column, and the use of the former as IS requires application of GC-MS. In the SIM mode, m/z 77 and 90 are suggested for d3-methyl esters of saturated fatty acids, whereas m/z 88 and 101 are recommended for ethyl esters of saturated fatty acids. These m/z values give either no or very low response for FAME and can thus be used for the analysis of FAME in food by GC-MS in the SIM mode. Fatty acids in sunflower oil and mozzarella cheese were quantified using five saturated FAEE as IS. Gravimetric studies showed that the transesterification procedure could be carried out without of loss of fatty acids. GC-EI/MS full scan analysis was suitable for the quantitative determination of all unsaturated fatty acids in both food samples, whereas GC-EI/MS in the SIM mode was particularly valuable for quantifying minor fatty acids. The novel GC-EI/MS/SIM method using fatty acid ethyl esters as internal standards can be used to quantify individual fatty acids only, that is, without determination of all fatty acids (the common 100% method), although this is present. This was demonstrated by the exclusive quantification of selected fatty acids including methyl-branched fatty acids, erucic acid (18:1n-9trans), and polyunsaturated fatty acids in cod liver oil and goat's milk fat.     

In vitro test for the effectiveness of antioxidants as inhibitors of thiyl radical-induced reactions with unsaturated fatty acids

Attacks of thiyl radicals on the double bonds of unsaturated fatty acids lead to stereomutation (cis-, trans-isomerization without double-bond migration) and addition reactions (thioether formation). On the basis of these findings, an in vitro test system has been developed which allows the study of the effectiveness of specific antioxidants in preventing thiyl radical-induced attacks on unsaturated fatty acids. The test involves thermal treatment of a mixture of oleic (cis-9-octadecenoic) acid and 1-tetradecanethiol with the antioxidant, followed by measurement of the extent of formation of the products of stereomutation and addition (i.e., elaidic (trans-9-octadecenoic) acid and isomeric 9(10)-S-tetradecylstearic acids, respectively) by gas chromatography of their methyl esters as a function of antioxidant concentration. Antioxidants such as octyl gallate, ascorbic acid 6-O-palmitate, ubiquinone 50 (coenzyme Q(10)), rac-alpha-tocopherol, and 2,6-di-tert-butyl-4-methylphenol (BHT) were tested for their ability to protect the >C=C< double bond of oleic acid against attacks of thiyl radicals generated from 1-tetradecanethiol by heating. The results show that octyl gallate, ascorbic acid 6-O-palmitate and, to some extent, ubiquinone 50 (coenzyme Q(10)) were highly effective in preventing reactions of free thiyl radicals with oleic acid, whereas rac-alpha-tocopherol and BHT were moderately effective.     

基于差示扫描量热法和热力学模型的生物柴油结晶行为分析

棕榈酸甲酯(C16:0)、硬脂酸甲酯(C18:0)和油酸甲酯(C18:1)是生物柴油的主要组成部分。为了深入探究生物柴油的结晶行为,该文基于差示扫描量热法(differential scanning calorimetry,DSC)分析了这3种脂肪酸酯的物性参数,研究发现饱和脂肪酸甲酯C16:0和C18:0的熔点和熔化焓远远高出不饱和脂肪酸甲酯C18:1的值,C16:0和C18:0的熔点分别为301.57、310.92 K,C18:1的熔点为255.01 K。对脂肪酸酯组成的二元溶液进行DSC扫描,DSC曲线出现了2个放热峰,并且溶液的结晶点要低于首先析出的饱和脂肪酸酯纯物质时的熔点;随着饱和脂肪酸酯质量分数的增加溶液的结晶点温度也相应提高。将生物柴油当作由多元脂肪酸甲酯的混合溶液时,C16:0和C18:0等饱和脂肪酸甲酯作为溶质,C18:1等不饱和脂肪酸甲酯作为溶剂,建立了热力学模型计算溶液的结晶点温度。将脂肪酸甲酯的混合溶液近似为理想溶液时对此模型进一步简化,并利用简化模型计算得到4种生物柴油的结晶温度,与实测值进行比较得到了很好的验证效果。该研究可为优化生物柴油低温流动性的技术措施提供参考。     

Effects of season and processing on oil content and fatty acids of baltic herring (Clupea harengus membras)

Fatty acid composition, oil content, free fatty acid content, and peroxide value of Baltic herring (Clupea harengus membras) and two processed products (fried fillets and fish burgers) were investigated. The highest oil content of the fillets was found in autumn (10%), at the time when the free fatty acids had their minimum (1.4%). The main fatty acids were oleic (18-23%), palmitic (17%), palmitoleic (8-12%), and docosahexaeneoic (8-10%) acids. The proportion of saturated fatty acids was a constant 23% all year around, whereas mono- and polyunsaturated acids varied from 34 to 39% and 33 to 37%, respectively. During processing the oil content doubled and the fatty acid composition changed to the pattern of the rapeseed oil used for frying. Oleic acid was a major fatty acid in the products comprising over 40% of the total fatty acids. The proportion of n-3 acids decreased during processing but the total amount of polyunsaturated acids remained fairly constant.     

NMR-based metabolomics reveals that conjugated double bond content and lipid storage efficiency in HepG2 cells are affected by fatty acid cis/trans configuration and chain length

In the present study the metabolic response to various fatty acids was investigated in HepG2 cells by using a (1)H NMR-based approach. To elucidate the effect of cis/trans configuration, the cells were exposed to either oleic acid (C18:1 cis-9), elaidic acid (C18:1 trans-9), vaccenic acid (C18:1 trans-11), linoleic acid (C18:2), or palmitic acid (C16:0), and multivariate data analysis revealed a strong effect of fatty acid on the lipophilic metabolite fraction. Inspection of the spectra revealed that the difference between the observed responses could be ascribed to the appearance of resonances from conjugated double bonds (5.65, 5.94, and 6.28 ppm) in cells exposed to vaccenic acid, revealing that vaccenic acid upon uptake by the HepG2 cells is converted into a conjugated fatty acid. Upon exposure of the HepG2 cells to either butyric acid (C4:0), caproic acid (C6:0), lauric acid (C12:0), myristic acid (C14:0), or palmitic acid (C16:0), an effect of fatty acid length was also evident, and data indicated that short-chain fatty acids (C4-C6) are immediately converted, whereas medium-long-chain fatty acids (C12-16) are incorporated into triglycerides and deposited in the cells. In conclusion, the present study demonstrates that (1)H NMR spectroscopy is a useful method for studying the uptake of fatty acids in in vitro cells.     

Identification of fatty acid esters of pectenotoxin-2 seco acid in blue mussels (Mytilus edulis) from Ireland

Pectenotoxins from marine dinoflagellates of the genus Dinophysis are rapidly hydrolyzed by many shellfish to give pectenotoxin-2 seco acid, which isomerizes to 7-epi-pectenotoxin-2 seco acid. Three series of fatty acid esters of pectenotoxin-2 seco acid (PTX-2 seco acid) and 7-epi-PTX-2 seco acid were detected by LC-MS analysis of extracts from blue mussels (Mytilus edulis) from Ireland. The locations of the fatty acid ester linkages were identified by a combination of LC-MSn in positive- and negative-ion modes, LC-MS analysis of the products from reaction of the esters with sodium periodate, and NMR analysis of purified samples of the two most abundant ester derivatives. The 37-O-acyl esters of PTX-2 seco acid were the most abundant, followed by the corresponding 11-O-acyl esters, accompanied by low levels of the 33-O-acyl esters. The most abundant fatty acid esters in the fractionated sample were, in order, the 16:0, 22:6, 14:0, 16:1, 18:4, and 20:5 fatty acids, although a wide array of other PTX-2 seco acid fatty acid esters were also present at low levels.     

C(18) acetylenic fatty acids of Ximenia americana with potential pesticidal activity

Bioactivity-driven fractionation of the CHCl(3) extract of the root of Ximenia americana, using the brine shrimp lethality test (BST) and hatchability test with Clavigralla tomentosicollis eggs, gave C(18) acetylenic fatty acids 1 and 2. 1 is octadeca-5-ynoic acid (tariric acid). 2 is a novel ene-ene-yne-ene acetylenic fatty acid (10Z,14E,16E-octadeca-10,14,16-triene-12-ynoi c acid). The structures of 1 and 2 were assigned from the MS and NMR data. Fractions that are rich in acetylenic fatty acids inhibited the hatching of C. tomentosicollis eggs.     

Identification and quantification of astaxanthin esters in shrimp (Pandalus borealis) and in a microalga (Haematococcus pluvialis) by liquid chromatography-mass spectrometry using negative ion atmospheric pressure chemical ionization

Negative ion liquid chromatography-atmospheric pressure chemical ionization mass spectrometry [negative ion LC-(APCI)MS] was used for the identification of astaxanthin esters in extracts of commercial shrimp (Pandalus borealis) and dried microalga (Haematococcus pluvialis) samples. A cleanup step using a normal phase solid phase extraction (SPE) cartridge was applied prior to analysis. Recovery experiments with astaxanthin oleate as model compound proved the applicability of this step (98.5 +/- 7.6%; n = 4). The assignment of astaxanthin esters in negative ion LC-(APCI)MS was based on the detection of the molecular ion (M*-) and the formation of characteristic fragment ions, resulting from the loss of one or two fatty acids. Quantification of individual astaxanthin esters was performed using an astaxanthin calibration curve, which was found to be linear over the required range (1-51 micromol/L; r2 = 0.9996). Detection limits, based on the intensity of M*-, a signal-to-noise ratio of 3:1, and an injection volume of 20 microL, were estimated to be 0.05 microg/mL (free astaxanthin), 0.28 microg/mL (astaxanthin-C16:0), and 0.78 microg/mL (astaxanthin-C16:0/C16:0), respectively. This LC-(APCI)MS method allows for the first time the characterization of native astaxanthin esters in P. borealis and H. pluvialis without using time-consuming isolation steps with subsequent gas chromatographic analyses of fatty acid methyl esters. The results suggest that the pattern of astaxanthin-bound polyunsaturated fatty acids of P. borealis does not reflect the respective fatty acid pattern found in triacylglycerides. Application of the presented LC-(APCI)MS technique in common astaxanthin ester analysis will forestall erroneous xanthophyll ester assignment in natural sources.     

Biosynthesis of trans-2-hexenal in response to wounding in strawberry fruit

Wounded strawberry fruit produces a diverse group of volatile compounds including aldehydes, alcohols, and esters derived from the lipoxygenase (LOX) and hydroperoxide lyase (HPL) pathways. Because the wound volatiles may play an important role in plant-fungal interaction, the goal of this study was to develop a greater understanding about the biosynthesis of the major wound volatile, trans-2-hexenal (t-2-H), produced by strawberry fruit upon wounding. To that end, composition and quantity of total and free fatty acids of control and wounded strawberry fruit were analyzed. In addition, activities of the key enzymes, LOX and HPL, and production of C6 aldehydes were determined. Intact strawberry fruit did not produce detectable t-2-H which is derived from alpha-linolenic acid (18:3). However, in response to wounding by bruising, strawberry fruit emitted t-2-H and its precursor cis-3-hexenal (c-3-H). The level of total lipid 18:3 in the fruit increased 2-fold in response to wounding, whereas free 18:3 declined slightly ( approximately 30%). At 10 min following wounding, fruit exhibited a 25% increase in LOX activity, which leads to the production of 13-hydroperoxyoctadecatrienoic acid (13-HPOT) from 18:3. The activity of HPL, which catalyzes formation of cis-3-hexenal from 13-HPOT, increased 2-fold by 10 min after wounding. Thus, during a 15 min period after wounding, free 18:3 substrate availability and the activity of two key enzymes, LOX and HPL, changed in a manner consistent with increased c-3-H and t-2-H biosynthesis.