Jonathan M. Curtis

CHARACTERIZATION OF BIOLOGICALLY INACTIVE SPIROLIDES E AND F : IDENTIFICATION OF THE SPIROLIDE PHARMACOPHORE

Abstract Two new spirolide derivatives, E and F, have been isolated in low yield from shellfish extracts. Absence of activity in the mouse bioassay of these derivatives, and of the secondary amine reduction product of spirolide B, identifies the spirolide pharmacophore as the cyclic imine moiety.

Two new water-soluble dsp toxin derivatives from the dinoflagellate prorocentrum maculosum: possible storage and excretion products

Two novel water-soluble sulfated DSP toxin derivatives 4 and 5, are reported from the benthic dinoflagellate Prorocentrum maculosum. Their occurrence supports the idea that all DSP toxin-producing Prorocentrum species biosynthesize such compounds as a means of toxin storage, and perhaps as a means of eventually exporting them from the cell.

Sulfoquinovosyl diacylglycerols from the alga Heterosigma carterae

An extract of the chloromonad Heterosigma carterae (Raphidophyceae), cultivated in natural seawater, contained a complex mixture of sulfoquinovosyl diacylglycerols. Palmitoyl (16:0), three isomers of hexadecenoyl (16:1 cis Δ9, Δ11, Δ13), and eicosapentenoyl (20:5) were found to be the main fatty acyl substituents. Exact double-bond sites were determined by mass spectrometry analysis of the corresponding nicotinyl derivatives. Four major sulfoquinovosyl diacylglycerol components were partially purified and identified as 1–4 by interpretation of their nuclear magnetic resonance and mass spectral data. In addition, complete analysis of the H. carterae sulfoquinovosyl diacylglycerols was performed using high-performance liquid chromatography combined with electrospray tandem mass spectrometry.

Preparation and certification of solutions of perdeuterated polycyclic aromatic compounds intended for use as surrogate internal standards

Two standard solutions of deuterated polycyclic aromatic compounds (PACs) have been prepared for use as surrogate internal standards. Solution DPAC-1 contains 21 deuterated PACs, and is intended for use with mass spectrometric (MS) detection. Most of the difficulties in certifying concentrations in DPAC-1 arose from the fact that none of the individual compounds was 100% deuterated, so that effects of mass spectrometric fragmentation are convoluted with those of isotopic distributions. The best methods are discussed for using such internal standards so as to minimize these problems, together with those arising from kinetic isotope effects. Solution DPAC-2 contains 6 deuterated PACs, and is primarily intended for use with reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection (FLD, dual programmed wavelength mode), in which the signals for analyte and internal standard are separated chromatographically rather than via the detector. Full details of the preparation of these solutions are described. In addition, examples of their use in the analysis of a certified coal-tar extract (NIST SRM 1597) are described briefly. In one example a novel HPLC-MS technique was employed, and in the other the HPLC-FLD technique was used.

Effects of ionization mode on charge-site-remote and related fragmentation reactions of long-chain quaternary ammonium ions.

Comparison of collisionally activated fragment spectra of long-chain quaternary ammonium ions, formed by liquid-assisted secondary ion mass spectrometry (LSIMS) and electrospray ionization (ESI), shows the latter are dominated by radical cations while the former yield mainly even-electron charge-site-remote (CSR) fragments, similar to the report for different precursors by Cheng et al., J. Am. Soc. Mass Spectrom.1998, 9, 840. Here, mixed-site fragmentation products (formal loss of a radical directly bonded to the nitrogen plus a radical derived from the long chain) are of comparable importance for both ionization techniques. These observations are difficult to understand if the CSR ions are formed by a concerted rearrangement-elimination reaction, since precollision internal energies of the ESI ions are much lower than those of the ions from LSIMS. Alternatively, if one discards the concerted mechanism for high-energy CA, and assumes that the even-electron fragments are predominantly formed via homolytic bond cleavage, the colder radical cations from ESI survive to the detector while the more energized counterparts from LSIMS preferentially lose a hydrogen atom to yield the CSR ions, as proposed by Wysocki and Ross (Int. J. Mass Spectrom. Ion Processes1991, 104, 179). The present work also attempts to reconcile discrepancies involving critical energies and known structures for neutral fragments.

Hoffmanniolide: a novel macrolide from Prorocentrum hoffmannianum

Abstract Hoffmaniolide, a novel macrolide, was isolated and identified from the marine dinoflagellate Prorocentrum hoffmannianum .

MIXED-SITE VS. CHARGE-SITE-REMOTE FRAGMENTATION REACTIONS OF LONG-CHAIN QUATERNARY AMMONIUM IONS

Abstract The mixed-site fragmentation reactions discovered by Tuinman, Cook and Magid (J. Am. Soc. Mass Spectrom., 1 (1989) 85), as reaction channels competitive with charge-site-remote fragmentations, have been further investigated. Two complementary deuterium-labelled tetra-alkylammonium ions, (CH3)3N+C14D29 and (CD3)3N+C14H29, were investigated as precursor ions for MS/MS/MS experiments using a sector/time-of-flight hybrid instrument employing orthogonal acceleration into the time-of-flight analyzer. By comparing the second-generation fragment ion spectra originating from the two precursors, it was possible unambiguously to assign atomic compositions and also to make plausible structural assignments. The fragment spectra thus obtained for the first-generation product ions of the charge-site-remote reactions are consistent with previous conclusions concerning their structures. The first-generation product ions from the mixed-site fragmentations, for example (CH3)2N+(CD2)m, are competitive with their charge-site-remote counterparts for m = 4, 5 and 6 in a manner consistent with the anticipated stabilities of cyclic transition states. However, interpretation of the second-generation fragment spectra of the (CH3)2N+(CD2)m ions and their deuterium-labelled complements was best accomplished in terms of a postulated diradical ion intermediate.

Charge-remote fragmentation characteristics of functionalized alkanes in high-energy collision- induced dissociation

In this study we report on high-energy, collision-induced dissociation processes leading to charge-remote fragmentations, using three alkyl cations, namely n-hexadecylpyridinium, n-hexadecyltriphenylphosphonium and n-hexadecyltriethylammonium, each with and without (2)H(2)-labelling at the C(9) position of the hexadecyl chain. The characteristic patterns corresponding to the formal elimination of alkane elements were observed, and the (2)H(2)-labelling at C(9) clearly affected only one charge-remote fragment ion of the homologous series. However, in addition to the expected fragment ion containing only one deuterium atom, a significant ion retaining two deuterium atoms was observed. MS/MS/MS experiments demonstrated clearly that the latter ion showed partial deuteration around the charge site, the level of deuteration depending on the structure of the original precursor cation. These results can be interpreted in terms of two novel, distinct mechanisms, one of which involves an excited state in an aromatic ring. Mixed-site fragmentation (MSF) ions were also observed from the phosphonium and ammonium ion precursors. We believe that the observation of the MSF process occurring at an sp(2)-hybridized center in the phosphonium series has not been reported previously. It thus becomes apparent that high-energy collisions leading to charge-remote reactions in fact lead to a broad range of pathways. Copyright 2000 John Wiley & Sons, Ltd.

TETREUTREPTIA POMQUETENSIS (EUGLENOPHYTA), A PSYCHROPHILIC SPECIES: GROWTH AND FATTY ACID COMPOSITION

Tetreutreptia pomquetensis McLachlan, Fritz et Seguel, a quadriflagellated euglenoid isolated from a shallow‐water embayment of the southeastern Gulf of St. Lawrence, is a common winter species where habitats are ice covered for 4 to 5 months. Growth in culture, which was similar to that of many euryhaline species, was limited at normal seawater salinities to temperatures between ∼0° and 7° C, and the alga could not be maintained at 10° C; thus, it is a strict psychrophile. The maximum growth rate (μ≅ 0.35·days−1) was around 5° C and, given its size (∼100 μm length), high productivities are indicated. The major fatty acids (FAs) identified were 18:4ω3, 16:4ω3, 20:5ω3, and 16:0. The overall FA spectrum does not align closely to other euglenoids but may reflect more the low growth temperature. The presence of 18:5ω3 is unusual, as this FA had been known previously only from dinoflagellates and haptophytes. The two most abundant sulfoquinovosyl diacylglycerols (SQDGs) had pairings of [18:4/16:0] and [18:4/18:4] and together constituted 80% of the total SQDGs. The latter pairing is unique, not having been previously recognized. The FA data support the hypothesis that the degree of unsaturation is indicative of low temperatures and membrane integrity.

Oxidative transformation of a naturally occurring okadaic acid diol ester by the diatom Thalassiosira weissflogii

Abstract The diarrhetic shellfish poisoning (DSP) toxin okadaic acid ( 1 ) is found in extracts of the dinoflagellate Prorocentrum lima together with a suite of diol esters such as 2 . When 2 was added to the culture medium of the centric diatom Thalassiosira weissflogii , it was transformed into a range of products within three days. Three of these products, 3 , 4 and 5 , were isolated from the medium and identified as oxidation products 2 , suggesting that the diatom produces DSP metabolites of greater polarity as a means of detoxification. In addition, another diol ester product 6 was identified as a minor impurity of 2 .