Enantiomeric separation of triacylglycerols containing fatty acids with a ring (cyclofatty acids).

Triacylglycerols (TAGs) containing cyclofatty acids (cycloFAs) from two oilseeds of Sterculia foetida and Hydnocarpus wightiana were analysed using both reversed-phase (RP18) and chiral phase columns. TAGs were identified using high-resolution electrospray ionization mass spectrometry in the positive ion mode. Fifty-five molecular species of TAGs have been identified in sterculic oil, 27 of which contained at least one cyclopropenyl-FA (e.g., malvalic or sterculic acids). The structures of regioisomers and enantiomers were determined for five major TAGs with cyclopropenyl-FAs. One hundred thirty-six TAGs were identified in chaulmoogra oil, 71 of which contained at least one cyclopentenyl-FA (e.g., gorlic, chaulmoogric, and hydnocarpic acids, etc.). Furthermore, in three molecular species, regioisomers and enantiomers were identified using HPLC on a chiral phase column. Eight molecular species of TAGs were prepared through organic synthesis to facilitate the identification of enantiomers. Retention times of fatty acid-containing triacylglycerols with one ring and one double bond are very similar to triacylglycerols with a dienoic fatty acid, but elution times are shorter. For example, dimalvaloylpalmitate elutes earlier than dilinoleylpalmitate. The order of elution of TAGs on the chiral column differs. In TAGs with 2 degrees of unsaturation (ring and double bond, e.g. PStP-StPP-PPSt), the order of elution is symmetric-asymmetric-asymmetric TAGs. TAGs with 4 degrees of unsaturation (one ring and three double bonds or two rings and two double bonds) present a different pattern. When TAGs contain two rings and two double bonds, the order of elution TAGs is asymmetric-symmetric-asymmetric (StStP-StPSt-PStSt); when TAGs contain a ring and 3 double bonds, the elution order is symmetric-asymmetric-asymmetric TAGs (OStO-StOO-OOSt). In species with a higher degree of unsaturation (e.g., 5), the elution order of the TAGs is asymmetric-asymmetric-symmetric (e.g. CCO-OCC-COC).

1-MCP treatment affects peach fruit aroma metabolism as revealed by transcriptomics and metabolite analyses.

1-methylcyclopropene (1-MCP) negatively affects peach aroma but the underlying molecular basis remains elusive. In this study, transcriptomics and metabolite analyses were carried out to investigate the regulatory mechanisms of 1-MCP on peach aroma from different standpoints: fatty acid (FA) metabolism, ethylene signal transduction and lipoxygenase/ß-oxidation pathway during 20 °C storage. Results indicate that 1-MCP significantly postponed the ethylene climacteric peak appearance and reduced ethylene production through down-regulation of related biosynthesis and signal transduction genes including PpaSAMS1/2, PpaACS1/2, PpaACO1 together with PpaETR1/2, PpaERS1, PpaEIN4 and PpaCTR1. Decrease in the levels of FAs and PpaFADs was observed in treated fruit, except oleic acid and PpaFAD4/5, before day 5 of storage. In addition, 1-MCP-treated fruit also possessed higher levels of C6 aldehydes and alcohols and delayed the formation of volatile compounds characteristic of peach-like aroma by upregulation of PpaLOX1/2/3 and PpaHPL1 expression and down-regulation of PpaLOX5 expression. Our findings suggest that inhibition of peach-like volatiles and promotion of green-note volatiles by 1-MCP were associated with ethylene production and modulation of FA levels through transcriptional regulation.

Identification of cyclopropyl fatty acids in walnut (Juglans regia L.) oil.

AIM: Identification of cyclopropyl fatty acids in walnut oil. METHOD: GC/MS method was developed for the determination of eight cyclopropyl fatty acids in walnut (Juglans regia) oil. RESULTS: Monocyclopropane acids: methyl 9-cyclopropyl-nonanoate, 6,7-methylene-, 8,9-methylene-, 9,10-methylene-, 11,12-methylene octadecanoates, and dicyclic acid - methyl 9,10,12,13-dimethylene octadecanoate, tricyclic acid - methyl 9,10,12,13,15,16-trimethylene octadecanoate, and unsaturated - methyl 2-octylcyclopropene-1-octanoate were detected in walnut oil by GC-MS and their mass spectra studied. Four cyclic fatty acids were identified for the fist time in seed oils. CONCLUSIONS: Eight cyclopropyl fatty acids were detected in the Mediterranean nuts for the first time.

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.

Determination of trinexapac in wheat by liquid chromatography-electrospray ionization tandem mass spectrometry.

A quantitative and confirmatory method for the analysis of trinexapac (free acid metabolite of trinexapac-ethyl) in wheat is described. Residues were extracted from wheat with acetonitrile in aqueous phosphate buffer (pH 7) overnight. The extract was directly injected into the HPLC system. Chromatographic separation was achieved on an octadecylsilica column, and detection was performed by negative ion electrospray ionization tandem mass spectrometry. The precursor ion of trinexapac [M - H](-) at m/z 223 was subjected to collisional fragmentation with argon to yield two intense diagnostic product ions at m/z 135 and 179, respectively. Accuracy and specificity for routine analysis of trinexapac were demonstrated. The validated concentration range was 10-200 microg/kg based on a 0.10 g/mL wheat sample extract. Recoveries were within the range of 71-94%, with associated relative standard deviations better than 10%. The limit of detection for trinexapac in wheat was estimated at 5 microg/kg. The method has been applied to a survey of 100 samples of wheat. In 46% of the samples analyzed, a quantifiable amount of trinexapac was detected, ranging from 10 to 110 microg/kg. It has been demonstrated that analyses of trinexapac accurately reflect the total amount of residues of the plant growth regulator, trinexapac-ethyl, in the wheat samples following field application. No residues of the parent compound, trinexapac-ethyl, in wheat were detected.

[Studies on chemical constituents of the essential oil from Angelica formosana].

Essential oil from Angelica formosana Boiss was obtained by steam distillation. The chemical components of the oil were examined by means of capillary gas chromatography and gas chromatography-mass spectrometry. 23 of the 62 separated constituents were identified. The content of each component identified was determined by area normalization method. Of these, the content of 11 compounds was higher than 1%.

Variation of cyclopropenoid fatty acids in cottonseed lipids.

The proportions of the cyclopropenoid fatty acids (CPA) esters, malvalate and sterculate, varied little in lipids from individual cottonseeds. Coefficients of variation were 10% and 20% for seeds from a lock and 13 varieties, respectively. Within the seed, variations in CPA concentrations were very large. Cyclopropenoid fatty acid concentration in the lipids decreased from 28% in the root tip to 2% in the top of the axis, and to 0.02% in the portion of the cotyledons nearest to the hull. The axial portion was only ca. 5% of the kernel, yet it contained 75% of the CPA. Distribution of dihydrosterculic acid, the precursor of CPA, was similar to that of CPA. High concentrations of CPA were found in immature seeds, root tip and radicle of germinated seeds, and root tips of cotton plants.

Effect of cultural conditions on the fatty acid composition of Thiobacillus novellus.

The fatty acid content of Thiobacillus novellus was determined under various cultural conditions. Four fatty acids, C(16:0), C(18:0), C(18:1), and a C(19) cyclopropane acid (C(19:cyc)), generally accounted for 90 to 99% of the total acids. Phosphate concentration, temperature, culture agitation, and the presence of branch-chain precursors had no significant effect on cellular fatty acids. Autotrophically grown cells contained more saturated C(16) and C(18) acids than did heterotrophic ones, and the sum of the percentages of the C(18:1) and the C(19:cyc) acids was consistently higher in the heterotrophs. When the inorganic medium was supplemented with biotin, autotrophic cells produced more C(19:cyc) and much less C(18:1) than did autotrophs in unsupplemented medium. Heterotrophic cells grown with biotin also showed a marked reduction of the unsaturated acid and an increase in the cyclopropane acid, except when glutamatecitrate medium was employed, in which case the opposite effect was noted. Two different biotin-supplemented media yielded cells with 75 to 77% of the total fatty acids being the C(19) cyclopropane acid, one of the highest reported values for this class of acid.

Comparison of the fatty acids of proteolytic type B and nonproteolytic types E and F of Clostridium botulinum.

Lipids were extracted from vegetative cells and spores of Clostridium botulinum. The total lipids extracted averaged approximately 3.8% of the dry weight of vegetative cells and 2.5% of the dry weight of spores of types 61E, "F," and 115B. The fatty acids were analyzed in the form of their methyl esters by gas-liquid chromatography. Infrared spectroscopy, mercuric acetate fractionation, and silver nitratethin layer chromatography served as complementary means of analysis. The total fatty acids included straight chain, saturated, unsaturated, and cyclopropane acids. Hexadecanoic and tetradecanoic acids were the predominant acids in both the spores and vegetative cells. Together, they comprised over 50% of the total fatty acids. Unsaturated acids were the second major group. These were primarily 7,8-tetradecenoic, 9,10-hexadecenoic, 7,8-hexadecenoic, 11,12-octadecenoic, and 9,10-octadecenoic acids. Nonproteolytic types 61E and "F" possessed an 18-carbon diunsaturate, which was not found in the vegetative cells or spores of proteolytic type 115B. A mechanism for the synthesis of unsaturated and cyclopropane acids was proposed.