RESEARCH IN RETROSPECT:Some Biograffiti of a Journeyman Chemist.

[Figure: see text] blacksquare, square, filled Abstract Small effusive leaks in vacuum systems, as in Knudsen cells and classical molecular beam machines, are responsible for many contributions to science in the twentieth century. Beginning in the 1950s, big convective leaks have turned out to be even more powerful and versatile investigative tools. Forming supersonic free jets, they have greatly expanded molecular beam methods, become the cornerstone of cluster science and technology, rewritten the book on molecular spectroscopy, and are adding new dimension to mass spectrometry. This account is the story of one man's experiences as a bystander and participant in these developments.

Ultrasonically assisted electrospray ionization for LC/MS determination of nucleosides from a transfer RNA digest.

The use of an ultrasonic nebulizer to assist electrospray ionization mass spectrometry (ESI-MS) has been described and demonstrated with the analysis of a transfer RNA digest by microcolumn LC. The restricted range of mobile-phase compositions amenable to the electrospray process has traditionally placed a severe limitation on the types of LC applications that can be used with ESI-MS. For this reason, an ultrasonic nebulizer configured for LC has been developed that can generate the fine dispersion of liquid required for ESI-MS from any type of mobile phase. In the case presented here, a transfer RNA was enzymatically digested into its substituent nucleosides, which were then analyzed by microcolumn LC. The required mobile-phase gradient (beginning at 5% methanol) falls outside the solvent range that can be used with conventional electrospray. The ultrasonic nebulizer, however, resolves this problem. The fundamental behavior of the four most common nucleosides (cytidine, adenosine, guanosine, uridine) was studied, and conclusions concerning the effects of solution chemistry were drawn. Specifically, signal from the H+ adducts of these species seems to be strongly dependent on the pKa value. Also, effects from several source operating variables were examined. These included capillary exit voltage, drying and focusing gases, and nebulizer frequency. Performance was found to be consistent over a wide range (0-100% methanol) of mobile-phase compositions. The limit of detection for adenosine injected onto a microcolumn was found to be 100 amol. Finally, nucleosides from as little as 150 fmol of RNA (amount prior to digestion) could be detected.

Ion formation from charged droplets: Roles of geometry, energy, and time.

The formation of ions from the charged droplets produced in the several spray ionization techniques is viewed as an activated rate process involving field-assisted desorption, in accord with the ideas first set forth by Iribame and Thomson. The novel features of the present treatment are particularly relevant to the unique ability of electrospray ionization to transform large molecules in solution to free ions in the gas phase, with extensive multiple charging. These new features stem mainly from the realization that the spacing of charges on a desorbed ion must relate to the spacing of charges on the surface of the droplet whence it came. The consequences of this "rule" can account for the existence of maxima and minima in the number of charges on the ions of a particular species as well as the nature of the distribution of ions among the intervening charge states. They also explain the dependence of charge state on the configuration in solution of the parent molecule of the desorbed ion. In addition, they provide insight into the sequence in time at which ions in the various charge states leave an evaporating droplet.

An ion-storage time-of-flight mass spectrometer for analysis of electrospray ions.

A preliminary design and implementation of a novel approach to electrospray-mass spectrometry are described. Based on a time-of-flight mass analysis, the instrument provides several important advantages for on-line mass analysis: 1, simplicity, ease of use and low manufacturing cost; 2, rapid scan speed, yielding quasi-instantaneous full mass scans at repetition rates up to several kHz; 3, soft ionization and accurate mass determination of extremely large analyte molecules; 4, high sensitivity.

Electrospray ionization for mass spectrometry of large biomolecules.

Electrospray ionization has recently emerged as a powerful technique for producing intact ions in vacuo from large and complex species in solution. To an extent greater than has previously been possible with the more familiar "soft" ionization methods, this technique makes the power and elegance of mass spectrometric analysis applicable to the large and fragile polar molecules that play such vital roles in biological systems. The distinguishing features of electrospray spectra for large molecules are coherent sequences of peaks whose component ions are multiply charged, the ions of each peak differing by one charge from those of adjacent neighbors in the sequence. Spectra have been obtained for biopolymers including oligonucleotides and proteins, the latter having molecular weights up to 130,000, with as yet no evidence of an upper limit.

Angular distribution of fluorescence from liquids and monodispersed spheres by evanescent wave excitation.

Experimental data on the angular distribution of fluorescence from thick liquid dye layers excited by evanescent waves are found to agree well with Fresnel theory and with an effective thickness formulation. Qualitative agreement of theory with fluorescence data obtained from monodispersed spherical particles having diameters comparable to the wavelength of the incident evanescent radiation and impregnated with dye molecules is also attained. From our results, the optimum incident and observation angles and polarizations to enhance the contrast and SNR for inelastic ATR reemission spectroscopy applied to studying micron or submicron layers of particulates can be predicted.

Calibration of mass spectrometers for van der Waals dimers.

A stream tube from a small supersonic free jet in vacuo is passed successively through the ion source of a quadrupole mass spectrometer and into an enclosed ionization gauge which indicates total mass flux. By simultaneous measurements of dimer and monomer ion currents as well as total mass flux we are able to arrive at absolute values of the relative concentration of neutral monomer and dimer in the free jet. Results obtained with argon, carbon dioxide, and oxygen indicate substantial fragmentation of dimers in the ion source. Consequently, populations of neutral dimers in supersonic free jets of the kind widely used as beam sources may be much larger than previous studies have indicated.

Molecular Beams with Energies above One Electron Volt.

By using as sources supersonic jets of hydrogen or helium containing small concentrations of heavier molecules we have been able to obtain molecular beams with kinetic energies of the heavy molecules well into the range above I electron volt. A variety of molecules have been successfully accelerated. Intensities of 10(16) to 10(17) heavy molecules per steradian-second have been achieved at these high energies.