Eser Ayanoglu

Minor and trace sterols in marine invertebrates: VI. Occurrence and possible origins of sterols possessing unusually short hydrocarbon side chains☆

Abstract Sterols with biosynthetically unusually short side chains (fewer than eight carbon atoms expected for primary squalene cyclization products) have been identified in the extracts of numerous marine invertebrates. The structures of the short side chain and conventional side chain sterols have been determined for various species of Porifera and Coelenterata. Sterol structures were determined by comparison of their mass spectra and gas chromatographic retention times with those of authentic or synthetic samples. Evidence is presented supporting the natural occurrence of these compounds in the tissues of the marine invertebrates as opposed to formation by degradative processes during sample handling or laboratory work-up. The short side chain sterols were found to possess predominantly the androst-5-en-3β-ol nucleus with C-17 alkyl side chains ranging from zero to six carbon atoms. Concentrations of short side chain sterols range from trace levels to over 5% of the sterol mixture in various species. The possible origins of these short side chain sterols are evaluated in the light of current knowledge of sterol function, biosynthesis, dealkylation, microbial degradation, and autoxidation. Known sterol autoxidations are reviewed, and possible singlet oxygen and free radical mechanisms of sterol side chain autoxidation (at physiological temperatures) which may lead to sterols with shortened hydrocarbon side chain are suggested. The possible autoxidative generation of short side chain sterols from known marine sterols by the suggested mechanisms is evaluated through application of the REACT computer program. Predicted short side chains are tabulated for each parent marine sterol side chain and then compared with the compositions of the actual sterols found in the marine extracts examined. The possible natural environmental or in vivo autoxidative formation of the short side chain marine sterols is supported by these evaluations.

Phospholipid studies of marine organisms: III. New phospholipid fatty acids fromPetrosia ficiformis

The fatty acyl components of the phospholipids from the spongePetrosia ficiformis consisted predominantly of branched, especially iso and anteiso acids. The two major components of the complex mixture are the hitherto unknown Z,Z-25-methyl-5,9-hexacosadienoic and Z,Z-24-methyl-5,9-hexacosadienoic acids. Other unknown acids are: 7,13,16-docosatrienoic acid, 15-methyldocosanoic acid, 15-methyltricosanoic acid and 24-methyl-5,9-pentacosadienoic acid. Short branched-chain fatty acids, presumably of bacterial origin, are considered to be the possible bioprecursors of these novel phospholipid constituents. The major phospholipids were PE, PC, PG, PS and PI. The distribution of fatty acids among the phospholipid classes was also studied.

Phospholipid studies of marine organisms: V1 new α-methoxy acids fromHigginsia tethyoides

The phospholipids of the demospongeHigginsia tethyoides are shown to have at least 16 long-chain α-methoxy acids, which represent a new class of fatty acids. Among them are the saturated α-methoxy acids containing 19–24 carbon atoms. The monounsaturated compounds are 2-OMe-Δ17-24:1, 2-OMe-Δ18-25:1, 2-OMeΔ19-26:1 and 2-OMe-Δ21-28:1. The major diunsatured ones were shown to be 2-OMe-Δ5, 19-26:2 and 2-OMe-Δ7, 21-28:2. Small amounts of 2-OMe-23∶1, 2-OMe-26∶3; 2-OMe-27∶1 and 2-OMe-28∶3 were also encountered. Structures of the minor monounsaturated compounds were tentatively assigned as 2-methoxy-16-tricosenoic acid and 2-methoxy-20-heptacosenoic acids. The double bonds of the fatty acids show all-cis configuration. Circular dichroism measurements indicate an R-configuration for the α-methoxy acids. The major component of the total phospholipid acid mixture is 5,9,23-triacontatrienoic acid. Possible biosynthetic routes to these unusual phospholipid acids are discussed. The major phospholipids were phosphatidylethanolamine, phosphatidylglycerol and phosphatidylserine. The distribution of fatty acids among the phospholipids was also investigated.

Phospholipid studies of marine organisms: New branched fatty acids fromStrongylophora durissima

The phospholipids of the spongeStrongylophora durissima were analyzed. The major phospholipids present were phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylglycerol (PG) and phosphatidylinositol (PI). The major fatty acid components of the phospholipids consisted of short chain (C14−C19) and very long chain (C25−C30) “Demospongic” acids. Three novel branched Δ5 monounsaturated acids,Z-19-methyl-5-pentacosenoic,Z-19-methyl-5-hexacosenoic andZ-19-methyl-5-heptacosenoic acids were encountered in the sponge. The 3-saturated counterparts of these compounds, 19-methylpentacosanoic, 19-methylhexacosanoic and 19-methylheptacosanoic acids, as well as 19-methylpentacosanoic and 20-methyloctacosanoic acids also are hitherto undescribed acids present in the sponge. Trace amounts of 2 very long chain acids also were detected and their structures tentatively assigned as 19,21-dimethylheptacosanoic and 20,22-dimethyloctacosanoic acids. The distribution of these fatty acids according to phospholipid head groups also was described.

Double-bond location in long-chain polyunsaturated fatty acids by chemical lonization-mass spectrometry

For determination of the double-bond position in polyunsaturated C24–30 fatty acids from marine organisms, methoxy derivatives were prepared. Diagnostic mass spectral fragment as well as molecular ion intensities were obtained by adjusting the ion source optics in the presence of ammonia at a lower source pressure than used conventionally. A lower detection limit was observed compared to conventional methane chemical ionization, which is a more favorable condition for capillary gas chromatography. Analysis of fatty acids from the spongeCalyx niceaensis showed the double-bond position of 8 unsaturated fatty acids, including two new ones. In addition, structural proof is provided for the presence of a new cyclopropane-containing fatty acid: 19,20-methylene-hexacosanoic acid.

Phospholipid studies of marine organisms 4. (2R,21Z)-2-methoxy-21-octacosenoic acid, the first naturally occurring α-methoxy acid from a phospholipid

Abstract A novel fatty acid, (2 R ,21 Z )-2-methoxy-21-octacosenoic acid, was shown to be present in the phospholipids of the sponge Higginsia tethyoides .

Phospholipid studies of marine organisms 9. New brominateddemospongic acids from the phospholipids of two petrosia species

Abstract Two Novel long chain fatty acids, (5Z, 9Z)-6-bromo-25-methyl-5,9-hexacosadienoic and (5Z, 9Z)-6-bromo-24-methyl-5,9-hexacosadienoic acids were found in the phospholipids of Petrosia ficiformis and P. hebes . Their structures were elucidated with the help of CI-EI7MS and a homogeneous hydrogenation catalyst.

Mass spectral behavior and HPLC of some unusual molecular phospholipid species

The molecular species of the major phospholipids from the marine sponges Parasperella psila and Microciona prolifera were studied using chemical hydrolysis, enzymatic degradation and capillary gas chromatography (GC), high performance liquid chromatography (HPLC), desorption chemical ionization (DCI), fast atom bombardment (FAB) combined with collisionally activated decomposition (CAD) mass spectrometry. Two new solvent systems were developed for the isolation of these species from the sponges. Our investigations indicated the existence of unusual symmetrical phospholipids as major components. 1,2-Di-(5Z,9Z)-5,9-hexacosadienoyl-sn-glycero-3-phosph oethanolamine was found in both organisms, while 1,2-di(5Z,9Z,19Z)-5,9,19-hexacosatrienoyl-sn-gly cero-3-phosphoethanolamine was present in M. prolifera, 1,2-Di-(4Z,7Z,10Z,13Z,16Z,19Z)-4,7,1 0,13,16,19-docosahexaenoyl-sn-glycero-3- phosphocholine was the major molecular species in the PC fraction of M. prolifera.

New natural 2-acetoxy fatty acids using chemical ionization and electron impact mass spectrometry

The phospholipids of the spongePolymastia gleneni contain saturated long chain (C22–30)-acetoxy fatty acids. Their structures were assigned based on chromatographic and spectrometric data as well as comparison with a synthetic sample. The use of capillary gas chromatography combined with chemical ionization and electron impact mass spectrometry was instrumental in the eludication of structures, since only a very small amount of crude lipids was available.

High performance liquid chromatography and fast atom bombardment mass spectrometry of unusual branched and unsaturated phospholipid molecular species

The unusual symmetrical molecular species 1,2-di-3,7,11,15-tetramethylhexadecanoyl-sn-glycero-3-phosphoglyce rol, 1,2-di-5,8,11,14-docosatetraenoyl-sn-glycero-3-phosphocholine, 1,2-di-5,9,19-octacosatrienoyl-sn-glycero-3-phosphoethanolamine, and 1,2-di-5,9,23-triacontatrienoyl-sn-glycero-3-phosphoethanolamine were isolated from the marine sponges Axinella verrucosa, Higginsia tethyoides, Tethya aurantia and Aplysina fistularis by HPLC and studied by fast atom bombardment (FAB) mass spectrometry. In addition to molecular weights, branching and double bonds were located in the fatty acyl chains of the intact phospholipid molecules, using FAB either in a positive or negative mode. Some mass spectral results were obtained on enriched phospholipid fractions rather than pure molecular species using MS/MS.