Patterns and sources of leaf tannin variation in yellow birch (Betula allegheniensis) and sugar maple (Acer saccharum)

Leaves from forest-grown sugar maple (Acer saccharum Marsh) and yellow birch (Betula allegheniensis Britt.) trees were analyzed for four tannin measures (hydrolyzable and condensed tannins, total phenolics, and protein binding) at three times during the growing season. Fifteen-year-old half-sib sugar maples from four provenances, representing the geographical extremes of the sugar maple range and growing in a common garden, were examined for the same traits. We found no significant geographic or seed source component to variation in three of the four tannin measures. We found significant seasonal changes in both birch and maple leaf tannins. Withincanopy leaf tannin variation tended to obscure differences between trees in maple, but in birches between-tree differences in leaf tannin content were more readily found. We also found a significant negative correlation between leaf protein binding capacity and leaf wet weight.     

Fatty acid composition of some pine seed oils

The fatty acid composition of seeds from seven species of the genusPinus (P. pinaster, P. griffithii, P. pinea, P. koraiensis, P. sylvestris, P. mughus, andP. nigra) was established. Pine seeds are rich in oil (31–68% by weight) and contain several unusual polymethylene-interrupted unsaturated fatty acids with acis-5 ethylenic bond. These are thecis-5,cis-9 18:2,cis-5,cis-9,cis-12 18:3,cis-5,cis-11 20:2, andcis-5,cis-11,cis-14 20:3 acids, with a trace ofcis-5,cis-9,cis-12,cis-15 18:4 acid. Their percentage relative to total fatty acids varies from a low of 3.1% (P. pinea) to a high of 30.3% (P. sylvestris), depending on the species. The majorcis-5 double bond-containing acid is generally thecis-5,cis-9,cis-12 18:3 acid (pinolenic acid). In all species, linoleic acid represents approximately one-half the total fatty acids, whereas the content of oleic acid varies in the range 14–36% inversely to the sum of fatty acids containing acis-5 ethylenic bond. The easily available seeds fromP. koraiensis appear to be a good source of pinolenic acid: their oil content isca. 65%, and pinolenic represents about 15% of total fatty acids. These values appear to be rather constant.Pinus pinaster, which is grown on several thousand acres in the southwest of France, is an interesting source ofcis-5,cis-11,cis-14 20:3 acid (7% in the oil, which isca. 35% of the dehulled seed weight), an acid sharing in common three double bonds with arachidonic acid. Apparently,P. sylvestris seed oil contains the highest level ofcis-5 double bond-containing acids among pine seed oils that have ever been analyzed.     

Determination of petroselinic acid inUmbelliflorae seed oils by combined GC and13C NMR spectroscopy analysis

Synthetic triolein and tripetroselinin mixtures were examined by¹³C NMR spectroscopy, showing marked chemical shift differences of the olefinic carbon atoms. Peak height ratios were compared to weight values for quantitative determination of oleic and petroselinic acids in seed oils, since these two fatty acids are quantitated together by GC analysis. Values observed for NMR peak height ratios were fairly close and agreed well with weight ratios. From overall compositions of eleic and petroselinic acids obtained by GC and relative compositions given by¹³C NMR, petroselinic acid has been determined in tenUmbelliflorae seed oils.     

Cis-trans isomerization of octadecatrienoic acids during heating. Study of pinolenic (cis-5,cis-9,cis-12 18∶3) acid geometrical isomers in heated pine seed oil

To understand the heat-inducedcis-trans isomerization of ethylenic bonds in octadecatrienoic acids, pine seed oil, which contains the unusual nonmethylene-interrupted pinolenic (cis-5,cis-9,cis-12 18∶3) acid as a major component, was heated under vacuum at 240°C for 6 h together with linseed and borage oils. As a results, a small percentage of pinolenic acid undergoescis-trans isomerization. The main isomer that accumulates is thetrans-5,cis-9,trans-12 18∶3 acid. Minor amounts of the three mono-trans isomers are also present. Identification of isomers was realized by combining gas-liquid chromatography on a CP Sil 88 capillary column, argentation thin-layer chromatography and comparing the equivalent chainlengths of artifacts to those of isomers present in NO₂-isomerized pine seed oil. Hydrazine reduction was used to demonstrate that there was no positional shift of double bonds. Heat-induced geometrical isomerization of pinolenic acid differs from that of α- and γ-linolenic acids in at least two aspects. The reaction rate is slower (about one-fourth), and mono-trans isomers are formed in low amounts.     

FA and unsaponifiable composition of five Amazonian palm kernel oils

The kernel oils of five different palm species native to the Amazon basin and French Guyana were studied. Those studied were Acrocomia lasiospatha Wall., Astrocaryum vulgare C. Mart., Bactris gasipaes H.B.K., Elaeis oleifera (Kunth) Cortés, and Maximiliana maripa Drude. Lauric and myristic acids were found in all of the oils. Analysis of the unsaponifiable contents, especially the sterol and triterpene alcohol determinations, revealed the preponderance of sitosterol and the presence of two triterpene alcohols (cycloartenol and 24-methylenecycloartanol). Antioxidant (vitamin E) levels were present in small amounts, with the levels being more similar to olive than to palm oil.     

One-step methodology for the synthesis of FA picolinyl esters from intact lipids

Picolinyl derivatives are used for structural determination of FA by GC-MS. Although they provide reliable diagnostic fragments, the usual multistep methodologies applied for their preparation require TAG hydrolysis or acid chloride formation prior to picolinyl synthesis. These reaction conditions may result in the presence of artifact molecules in the samples and thus compromise analytical quality and accuracy. To address these problems, a rapid, simple and quantitative methodology for the synthesis of FA picolinyl esters from intact lipids was developed. It involves their transesterification under basecatalyzed conditions using 3-potassiooxamethylpyridine in methylene chloride. The catalyst was prepared by proton exchange between potassium tert-butoxide and anhydrous 3-hydroxymethylpyridine. Mild reaction conditions allowed complete derivatization of TAG and phospholipids in 2 min at room temperature, and of FAME in 15 min at 45°C. The proposed procedure, which can be used on a routine basis, was applied to Ipomoae imperialis seed lipids and used to confirm occurrence of γ-linoleic acid at a level of 0.9%.     

Fatty acid and triacylglycerol compositions of seed oils of five Amaranthus accessions and their comparison to other oils

This paper reports the fatty acid and triacylglycerol (TAG) compositions of five Amaranthus accessions (RRC1011, R149, A.K343, A.K432, and A. K433) representing two species and a cross between one of these and a third species. Seed oils of these were analyzed by gas chromatography and reversed-phase high-performance liquid chromatography, and their compositional properties compared with buck-wheat (Fagopyrum esculentum), corn (Zea mays), rice bran (Oryza sativa), soybean (Glycine max L. Merr.), sesame (Sesamum indicum), quinoa (Chenopodium quinoa), and cottonseed (Gossypium hirsutum) oils. All Amaranthus accessions were relatively high in palmitic (21.4–23.8%) and low in oleic (22.8–31.5%) and linolenic (0.65–0.93%) acids when compared to most of the grain and seed oils. The fatty acid composition of Amaranthus accessions K343, K433, and K432 (group I) were different from R149 and RRC1011 (group II) in mono and polyunsaturated fatty acids, but the saturate/unsaturate (S/U) ratios were very similar. All Amaranthus accessions were similar in TAG type, but showed slight differences in percentage. High similarities in UUU, UUS, and USS composition were observed among Amaranthus K343, K433 and K432, and between R149 and RRC1011. The fatty acid compositions of Amaranthus oil (group I) and cottonseed oil were similar, but their TAG compositions were different. The grain and oilseed oils were different from each other and from the Amaranthus accessions oils in terms of fatty acid composition, S/U, and TAG ratios. The UUU, UUS, and USS percentages were very diverse in grain and seed oils. The percentages of squalene in the TAG sample from the Amaranthus accessions were 8.05% in K343, 11.10% in K433, 11.19% in K432, 9.96% in R149, and 9.16% in RRC1011. Squalene was also tentatively identified in quinoa and ricebran oils at levels of 3.39 and 3.10%, respectively.     

Characterization of γ-linolenic acid geometrical isomers in borage oil subjected to heat treatments (deodorization)

Heating of borage oil, either under vacuum as a model or during steam-vacuum deodorization, produces artifacts that are geometrical isomers of γ-linolenic acid (cis-6,cis-9,cis-12 18∶3 acid). In a first approach, we have studied the behavior of these fatty acids in the form of either methyl or isopropyl esters on two capillary columns (CP-Sil 88 and DB-Wax). From this study, it appears that the DB-Wax capillary column is the best suited analytical tool to study in some detail γ-linolenic acid geometrical isomers. In a second approach, the structure of these isomers was formally established by combining several analytical techniques: Argentation thin-layer chromatography, comparison of the equivalent chainlengths with those of isomers present in NO₂-isomerized borage oil on two different capillary columns, partial hydrazine reduction, oxidative ozonolysis, gas chromatography coupled with mass spectrometry and gas chromatography coupled with Fourier transform infrared spectroscopy. The two main isomers that accumulate upon heat treatments are thetrans-6,cis-9,cis-12 andcis-6,cis-9,trans-12 18∶3 acids with minor amounts ofcis-6,trans-9,cis-12 18∶3 acid. One di-trans isomer, supposed to be thetrans-6,cis-9,trans-12 18∶3 acid, is present in low although noticeable amounts in some of the heated oils. The content of these artificial fatty acids increases with increasing temperatures and duration of heating. The degree of isomerization (DI) of γ-linolenic acid is less than 1% when the oil is deodorized at 200°C for 2 h. Heating at 260°C for 5 h increases the DI up to 74%. Isomerization of γ-linolenic acid resembles that of α-linolenic (cis-9,cis-12,cis-15 18∶3) acid in several aspects: The same kinds and numbers of isomers are formed, and similar degrees of isomerization are reached when the octadecatrienoic acids are heated under identical conditions. It seems that the reactivity of a double-bondvis-à-vis cis-trans isomerization is linked to its relative position, central or external, and not to its absolute position (Δ6, 9, 12 or 15).     

Differential scanning calorimetric analysis of edible oils: Comparison of thermal properties and chemical composition

The thermal profiles of 17 edible oil samples from different plant origins were examined by differential scanning calorimetry (DSC). Two other confirmatory analytical techniques, namely gas-liquid chromatography (GLC) and high-performance liquid chromatography (HPLC), were used to determine fatty acid (FA) and triacylglycerol (TAG) compositions. The FA and TAG compositions were used to complement the DSC data. Iodine value (IV) analysis was carried out to measure the degree of unsaturation in these oil samples. The DSC melting and crystallization curves of the oil samples are reported. The contrasting DSC thermal curves provide a way of distinguishing among these oil samples. Generally, the oil samples with a high degree of saturation (IV<65) showed DSC melting and crystallization profiles at higher temperature regions than the oil samples with high degree of unsaturation (IV>65). Each thermal curve was used to determine three DSC parameters, namely, onset temperature (T _o ), offset temperature (T _f ) and temperature range (difference between T _o and T _f ). Reproducibility of DSC curves was evaluated based on these parameters. Satisfactory reproducibility was achieved for quantitation of these DSC parameters. The results show that T _o of the crystallization curve and T _f of the melting curve differed significantly (P<0.01) in all oil samples. Our observations strengthen the premise that DSC is an efficient and accurate method for characterizing edible oils.     

13C nuclear magnetic resonance spectroscopic analysis of the triacylglycerol composition ofBiota orientalis and carrot seed oil

¹³C nuclear magnetic resonance (NMR) spectroscopic analysis of the whole oil (triacylglycerols) ofBiota orientalis seeds confirms the presence of oleate [18:1(9Z)], linoleate [18:2(9Z, 12Z)], linolenate [18:3((9Z, 12Z, 15Z)], 20:3 (5Z, 11Z, 14Z), 20:4(5Z, 11Z, 14Z, 17Z), and saturated fatty acids in the acyl groups by comparing the observed carbon shifts with previously established shift data for model triacylglycerols. This technique shows that the saturated, 20:3 and 20:4 fatty acids are distributed mainly in the α-acyl positions, whereas oleate, linoleate, and linolenate are randomly acylated to the α- and β-positions of the glycerol “backbone”. Stereospecific hydrolysis of theBiota oil with pancreatic lipase, followed by chromatographic analysis of fatty esters, reveals the presence of trace amounts of 16:0(0.7%), 18:0(0.5%), 20:3 (0.4%), and 20:4 (1.3%) in the β-position of the glycerol “backbone”, which are undetectable by¹³C NMR technique on the whole oil. Semiquantitative assessment of the¹³C NMR signal intensities gives the relative percentages of the fatty acid distribution as: saturated 16:0, 18:0 (12.0% α-acyl), oleate (7.7% α-acyl 8.7% β-acyl), total linoleate and linolenate (31.7% α-acyl; 24.2% βacyl), total 20:3 and 20:4 (15.7% α-acyl). The¹³C NMR spectroscopic analysis of carrot seed oil identifies the presence of saturated (18:0), 18:1(6Z), 18:1(9Z), and 18:2(9Z, 12Z). The saturated fatty acid is found in the α-acyl positions. Semi-quantitative assessment of the signal intensities gives the relative percentages of the fatty acids as: 18:0 (4.5% α-acyl), 18:1(6Z) (49.6% α-acyl; 19.7% β-acyl), oleate (6.5% α-acyl; 8.6% β-acyl) and linoleate (5.2% α-acyl; 6.9% β-acyl).     

Fatty acid (FA) composition and contents of <Emphasis Type="Italic">trans</Emphasis> unsaturated FA in hydrogenated vegetable oils and blended fats from Pakistan

This study presents the FA composition and trans FA (TFA) contents of different hydrogenated vegetable oils and blended fats marketed in Pakistan. Thirty-four vanaspati (vegetable ghee), 11 shortenings, and 11 margarines were analyzed. The contents of saturated FA, cis monounsaturated FA, and cis PUFA were in the following ranges: vanaspati 27.8–49.5, 22.2–27.5, 9.3–13.1%; vegetable shortenings 37.1–55.5, 15.8–36.0, 2.7–7.0%; and margarines 44.2–55.8, 21.7–39.9, 2.9–20.5%, respectively. Results showed significantly higher amounts of TFA in vanaspati samples, from 14.2 to 34.3%. Shortenings contained TFA proportions of 7.3–31.7%. The contents of TFA in hard-type margarines were in the range of 1.6–23.1%, whereas soft margarines contained less than 4.1% TFA.     

An efficient method for the purification of arachidonic acid from fungal single-cell oil (ARASCO)

PUFA, such as arachidonic acid (AA), have several pharmaceutical applications. An efficient method was developed to obtain high-purity arachidonic acid (AA) from ARASCO, a single-cell oil from Martek (Columbia, MD). The method comprises three steps. In the first step, AA was enriched from saponified ARASCO oil by low-temperature solvent crystallization using a polar, aprotic solvent, which gave a FA fraction containing 75.7% AA with 97.3% yield. The second step involved enriching AA content via lipase-catalyzed selective esterification of FA with lauryl alcohol. When a mixture of 1 g FA/lauryl alcohol (2∶1 mol/mol), 50 mg Candida rugosa lipase, and 0.33 g water was incubated at 50°C for 24 h with stirring at 400 rpm, the AA content in the unesterified FA fraction was as much as 89.3%, with ca. 90% yield. Finally, a solvent extraction procedure, in which acetonitrile was the extracting solvent, was used to enrich AA from FA fraction dissolved in n-hexane. The best results were obtained when 2 g FA was dissolved in 80 mL hexane and extracted twice, each time with 20 mL acetonitrile at −20°C, by allowing 2 h storage. This step gave a FA fraction containing 95.3% AA with 81.2% yield. By using this three-step process the AA content in the saponified single-cell oil (ARASCO) was increased from 38.8 to 95.3% with a total yield of ca. 71%.     

Proportions of C18∶1n−7 and C18∶1n−9 fatty acids in canola seedcoat surface and internal lipids

Lipids of canola seedcoats (Brassica napus L. andB. rapa L.) were prepared by surface washing and by complete extraction of seed coats with toluene. The major fatty acyl-containing triacylglycerols, wax esters and free fatty acids were separated by thin-layer chromatography prior to transesterification and analysis by gas-liquid chromatography. The proportion of C18∶1n−7 to C18∶1n−9 was higher in the extracted lipids than in the surface-washed lipids for all three classes.     

Production of structured TAG rich in 1,3-dicapryloyl-2-γ-linolenoyl glycerol from borage oil

γ-Linolenic acid (GLA) has the physiological functions of modulating immune and inflammatory responses. We produced structured TAG rich in 1,3-dicapryloyl-2-γ-linolenoyl glycerol (CGC) from GLA-rich oil (GLA45 oil; GLA content, 45.4 wt%), which was prepared by hydrolysis of borage oil with Candida rugosa lipase having weak activity on GLA. A mixture of GLA45 oil/caprylic acid (CA) (1∶2, w/w) was continuously fed into a fixed-bed bioreactor (18×180 mm) packed with 15 g immobilized Rhizopus oryzae lipase at 30°C, and a flow rate of 4 g/h. The acidolysis proceeded efficiently, and a significant decrease of lipase activity was not observed in full-time operation for 1 mon. GLA45 oil contained 10.2 mol% MAG and 27.2 mol% DAG. However, the reaction converted the partial acylglycerols to structured TAG and tricaprylin and produced 44.5 mol% CGC based on the content of total acylglycerols. Not only FFA in the reaction mixture but also part of the tricaprylin and partial acylglycerols were removed by molecular distillation. The distillation resulted in an increase of the CGC content in the purified product to 52.6 mol%. The results showed that CGC-rich structured TAG can efficiently be produced by a two-step process comprising selective hydrolysis of borage oil using C. rugosa lipase (first step) and acidolysis of the resulting GLA-rich oil with CA using immobilized R. oryzae lipase (second step).     

γ-Linolenic and stearidonic acids in Mongolian Boraginaceae

Seeds of nine Central Asian species of Boraginaceae were investigated for the first time for their oil content and for the fatty acid composition of their seed oils by capillary gas chromatography. Levels of γ-linolenic acid ranged from 6.6 to 13.0% and levels of stearidonic acid ranged from 2.4 to 21.4% of total seed fatty acids. The seed oil ofHackelia deflexa exhibited the highest stearidonic acid content (21.4%) that has been found so far in nature. Other high contents of this fatty acid were in threeLappula species (17.2 to 18.1%). Seed oils ofCynoglossum divaricatum andAmblynotus rupestris contain considerable amounts ofcis-11-eicosenoic (5.3 to 5.8%) andcis-13-docosenoic acid (7.0 to 9.7%) besides γ-linolenic (10.2 to 13.0%) and stearidonic acid (2.4 to 6.5%), which distinguish these oils from those of other Boraginaceae genera. This paper was presented as a poster at 10th Minisymposium and Workshop on Plant Lipids, Sept. 3–6, 1995, in Berne, Switzerland.     

Enzymatic fractionation of fatty acids: Enrichment of γ-linolenic acid and docosahexaenoic acid by selective esterification catalyzed by lipases

Immobilized lipase preparations from seedlings of rape (Brassica napus L.) andMucor miehei (lipozyme) used as biocatalysts in esterification and hydrolysis reactions discriminate strongly against γ-linolenic and docosahexaenoic acids/acyl moieties. Utilizing this property, γ-linolenic acid contained in fatty acids of evening primrose oil has been enriched seven to nine-fold, from 9.5 to almost 85% by selective esterification of the other fatty acids with butanol. Similarly, docosahexaenoic acid of cod liver oil has been enriched four to five-fold, from 9.4 to 45% by selective esterification of fatty acids (other than docosahexaenoic acid) with butanol. As long as the reaction is stopped before reaching equilibrium, very little of either γ-linolenic acid or docosahexaenoic acid are converted to butyl esters, which results in high yields of these acids in the unesterified fatty acid fraction.     

Electrochemical synthesis and characterization of poly(2,2’-dithiodianiline) thin films loaded with copper microparticles. Application to the amperometric analysis of γ-aminobutyric acid

Summary Modified electrodes with poly(2,2’-dithiodianiline),PDTDA, were prepared from the corresponding monomer by cyclic voltammetry between 0.0 V and 1.5-1.6 V on gold or stainless steel respectively. The potentiodynamic method proved to bring about good quality and adherent thin films. The wide potential window shown by this modified electrode allowed attempting the insertion of copper in the polymeric matrix using several strategies. The response of this modified electrode copper was checked in the presence of γ-aminobutyric acid (GABA). Copper loaded on PDTDA/Au electrodes showed the highest sensitivity along with a stable and reproducible response for the detection of GABA. The surface morphology and composition analysis by SEM and XPS shows that copper is deposited in the polymeric matrix as uniformly scattered microparticles. The surface of these particles is mainly composed of Cu(I) species.     

A minor source of vernolic,malvalic, and sterculic acids inPithecollobium dulce (syn.Inga dulcis) seed oil

Pithecollobium dulce, Benth (syn.Inga dulcis, Willd) seed oil, belonging to the Leguminosae plant family, contains minor amounts of vernolic acid (12,13-epoxy-octadec-cis-9-enoic acid, 10.0%), malvalic acid [7-(2-octacyclopropen-1-yl)heptanoic acid, 3.2%], and sterculic acid [8-(2-octacyclopropen-1-yl)octanoic acid, 2.0%]. The other normal fatty acids are palmitic (12.1%), stearic (4.2%), behenic (10.6%), oleic (34.1%), and linoleic (23.8%). These fatty acids have been characterized by Fourier transform infrared,¹H nuclear magnetic resonance, mass spectrometry and gas-liquid chromatography techniques and by chemical degradations.     

Positional distribution of 24∶6(n−3) in triacyl-sn-glycerols from flathead flounder liver and flesh

This paper presents the positional distribution of very long-chain fatty acids, 24∶6(n−3), in triacyl-sn-glycerols (TG) of flathead flounder (Hippoglossoides dubius). Each of the liver and flesh TGs was subjected to the stereospecific analysis. The liver TGs contained 24∶6(n−3) at concentrations of 1.5, 1.2 and 1.7 mole % in thesn-1,sn-2 andsn-3 positions, respectively, and the flesh TGs had 9.0, 7.8 and 7.1 mole % in thesn-1,sn-2 andsn-3 positions, respectively. This fatty acid was distributed almost evenly among the three positions of the TGs. No preference for thesn-2 position was observed in contrast to the general tendency for the distribution of longer-chain polyunsaturated fatty acids, such as 22∶6(n−3), 22∶5(n−3) and 20∶5(n−3). There was essentially no difference in the positional distributions of the liver and flesh TGs. The results obtained in this study give new fundamental information to the investigation of very long-chain fatty acids.     

Evaluation of the CP-Sil 88 and SP-2560 GC columns used in the recently approved AOCS official method Ce 1h-05: Determination of cis-, trans-, saturated, monounsaturated, and polyunsaturated fatty acids in vegetable or non-ruminant animal oils and fats by capillary GLC method

The AOCS Official Method Ce 1h-05 was recently approved at the 96th AOCS Annual Meeting (2005) by the Uniform Methods Committee as the official method for determining cis and trans FA in vegetable or non-ruminant fats and oils. A series of experiments was undertaken using a margarine (hydrogenated soybean oil) sample containing approximately 34% total trans FA (28% 18∶1 trans, 6% 18∶2 trans, and 0.2% 18∶3 trans), a low-trans oil (ca. 7% total trans FA), and a proposed system suitability mixture (12∶0, 9c−18∶1, 11c−18;1, 9c,12c,15c−18∶3, 11c−20∶1, and 21∶0) in an effort to evaluate and optimize the separation on the 100-m SP-2560 and CP-Sil 88 flexible fused-silica capillary GC columns recommended for the analysis. Different carrier gases and flow rates were used during the evaluation, which eventually lead to the final conditions to be used for AOCS Official Method Ce 1h-05.