Charge derivatization of peptides for analysis by mass spectrometry.

The analysis of peptide derivatives by fast atom bombardment, liquid secondary-ionization mass spectrometry, plasma desorption, electrospray ionization, and matrix-assisted laser desorption/ionization is reviewed. The fragmentation patterns of peptides and of charge-derivatized peptides are compared, and the proposed fragment ion structures are summarized. A variety of derivatization approaches and the distinguishing features of mass spectra produced from these derivatives are described. The most promising derivatization approaches are evaluated, and the strengths and limitations of these approaches are discussed.

A picomole-scale method for charge derivatization of peptides for sequence analysis by mass spectrometry.

A highly activated ester containing a fixed positive charge, S-pentafluorophenyl [tris(2,4,6-trimethoxyphenyl)phosphonium]acetate bromide (TMPP-AcSC6F5 bromide), has been synthesized as a reagent for N-terminal modification of peptides. Stable in aqueous acetonitrile solution during extended storage, TMPP-AcSC6F5 bromide reacts with unprotected peptides through p-(dimethylamino)pyridine (DMAP)-promoted amidation in aqueous acetonitrile (15 min, ambient temperature) to form N-TMPP-Ac derivatives of peptides. These peptide derivatives are readily amenable to analysis by fast atom bombardment (FAB) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Greater than 90% conversion has been observed in transforming low-nanomole quantities of analyte using molar ratios of 1:5:10 (peptide/reagent/ DMAP). For reactions at the picomole level a slightly modified stoichiometry, with molar ratios of 1:10:500, is employed. Owing to the high reaction efficiency and the tolerance to moderate excess reagent and base during analysis by FAB- and MALDI-MS, the reaction mixture containing the modified peptides can be analyzed directly in most cases, without sample cleanup. Examples of the preparation and analysis of a variety of N-TMPP-acetyl-peptides (TMPP-Ac-peptides) ranging from hexamers to 15-mers are given. Collisionally activated dissociation tandem mass spectrometry of TMPP-Ac-derivatives showed dominant a-type ions, accompanied by d- and c-type ions in some cases, allowing sequence determination to be made in a straightforward manner.

Investigation of the effects of solution composition and container material type on the loss of 11-nor-delta 9-THC-9-carboxylic acid.

The loss of 11-nor-delta 9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH) from solution was studied using fluorescence polarization immunoassay (FPIA) technology and x-ray photoelectron spectroscopy (XPS). Several materials (glass, silylated glass, high density polyethylene, polypropylene, polystyrene, polymethylmethacrylate, Teflon, and Kynar) were studied along with three solvents (water, urine, and Abbott cannabinoids diluent). THC-COOH losses ranging from 0 to 9.7 ng/cm2 and concentration reductions to 46% of starting values were measured. XPS indicated the presence of fluorine-labeled THC-COOH at materials surfaces. A half-life of 10 min was calculated for THC-COOH loss from urine stored in high density polyethylene at room temperature. Sample handling losses during pipetting were determined and ranged from 1.1 to 7.9 ng per aliquot. The effects of sample volume and sample handling on the THC-COOH concentrations of controls were also investigated.

Ab initio calculations on (Z)-5-decenyl acetate, a component of the pheromone complex of Agrotis segetum (Lepidoptera: Noctuidae) and electrophysiological studies with chain elongated analogues.

Conformational analyses of (Z)-5-decenylacetate, a sex pheromone component of the turnip moth, Agrotis segetum, and double unsaturated pheromone analogues 4 and 5 have been performed by ab initio calculations using Gaussian 92. Two minima were found for a cisoid and a transoid conformer, differing for 0.03 kcal/mol only. Conformational energies of diene analogues (5Z,7E)-5,7-decadienyl acetate (4) and (3E,5Z)-3,5,-decadienyl acetate (5) were determined for conformers required to mimic spatial relationships of the cisoid conformation of the natural pheromone 2. Finally, single sensillum recording studies were carried out with chain elongated C11- to C16-pheromone analogues 6.

The diffusion of L(+)-ascorbic acid across DPPC vesicle membranes.

The transmembrane diffusion of L(+)-ascorbic acid has been studied by means of 1H-n.m.r. spectroscopy using small unilamellar DPPC vesicles as a model system. It is shown that the electro-neutral form of L(+)-ascorbic acid diffuses faster across the membranes than the anionic form by about two orders of magnitude. The diffusion is influenced by a molecular interaction between L(+)-ascorbate and the membrane surface and depends also on the fluidity of the membrane. The calculated permeation coefficients of neutral L(+)-ascorbic acid are between 6 x 10(-10) and 3 x 10(-8) cm s-1 (35-52 degrees C).