Retinal shows its true colours: photoisomerization action spectra of mobility-selected isomers of the retinal protonated Schiff base.

Retinal is one of Nature's most important and widespread chromophores, exhibiting remarkable versatility in its function and spectral response, depending on its protein environment. Reliable spectroscopic and photochemical data for the isolated retinal molecule are essential for calibrating theoretical approaches that seek to model retinal's behaviour in complex protein environments. However, due to low densities and possible co-existence of multiple isomers, retinal is a challenging target for gas-phase investigations. Here, the photoisomerization behaviour of the trans isomer of the retinal protonated Schiff base (RPSB) is investigated in the gas phase by irradiating mobility-selected RPSB ions with tunable light in a tandem ion mobility spectrometer. trans RPSB ions are converted to single cis isomers and also more compact isomers through irradiation with visible light. The S1← S0 photoisomerization action spectrum of trans RPSB, obtained by monitoring production of cis isomers as a function of wavelength, exhibits a single well-defined peak with a maximum at 618 ± 5 nm. Corresponding action spectra of cis RPSB isomers exhibit broader peaks, conclusively demonstrating an isomeric dependence for the RPSB spectrum in the gas phase.

Photoisomerization action spectrum of retinal protonated Schiff base in the gas phase.

The photophysical behaviour of the isolated retinal protonated n-butylamine Schiff base (RPSB) is investigated in the gas phase using a combination of ion mobility spectrometry and laser spectroscopy. The RPSB cations are introduced by electrospray ionisation into an ion mobility mass spectrometer where they are exposed to tunable laser radiation in the region of the S1 ← S0 transition (420-680 nm range). Four peaks are observed in the arrival time distribution of the RPSB ions. On the basis of predicted collision cross sections with nitrogen gas, the dominant peak is assigned to the all-trans isomer, whereas the subsidiary peaks are assigned to various single, double and triple cis geometric isomers. RPSB ions that absorb laser radiation undergo photoisomerization, leading to a detectable change in their drift speed. By monitoring the photoisomer signal as a function of laser wavelength an action spectrum, extending from 480 to 660 nm with a clear peak at 615 ± 5 nm, is obtained. The photoisomerization action spectrum is related to the absorption spectrum of isolated retinal RPSB molecules and should help benchmark future electronic structure calculations.

Ion Mobility Unlocks the Photofragmentation Mechanism of Retinal Protonated Schiff Base.

Retinal protonated Schiff base (RPSB) is a key molecular component of biological photoreceptors and bacterial photosynthetic structures, where its action involves photoisomerization around bonds in the polyene chain. In a vacuum environment, collisional activation or exposure to visible light causes the RPSB molecule to disintegrate, producing charged molecular fragments with m/z = 248 Da that cannot be formed by simple cleavage of the polyene chain. Photofragments resulting from laser excitation of RPSB at a wavelength of 532 nm are analyzed in an ion mobility mass spectrometer (IMMS) and found to be the protonated Schiff base of ß-ionone. Density functional theory calculations at the M06-2X/cc-pVDZ level support a fragmentation mechanism in which RPSB undergoes an electrocyclization/fragmentation cascade with the production of protonated Schiff base of ß-ionone and toluene.

An ion mobility mass spectrometer for investigating photoisomerization and photodissociation of molecular ions.

An ion mobility mass spectrometry apparatus for investigating the photoisomerization and photodissociation of electrosprayed molecular ions in the gas phase is described. The device consists of a drift tube mobility spectrometer, with access for a laser beam that intercepts the drifting ion packet either coaxially or transversely, followed by a quadrupole mass filter. An ion gate halfway along the drift region allows the instrument to be used as a tandem ion mobility spectrometer, enabling mobility selection of ions prior to irradiation, with the photoisomer ions being separated over the second half of the drift tube. The utility of the device is illustrated with photoisomerization and photodissociation action spectra of carbocyanine molecular cations. The mobility resolution of the device for singly charged ions is typically 80 and it has a mass range of 100-440 Da, with the lower limit determined by the drive frequency for the ion funnels, and the upper limit by the quadrupole mass filter.

Changing the shape of molecular ions: photoisomerization action spectroscopy in the gas phase.

A new approach for studying the photoisomerization of molecular ions in the gas phase is described. Packets of molecular ions are injected into a drift tube filled with helium buffer gas, where they are irradiated with tunable laser light. Photoisomerization changes the ions' cross section for collisions with helium atoms so that they arrive at the ion detector slightly earlier or later than the parent ions. By monitoring the photo-isomer peak as a function of laser wavelength one can record an action spectrum that is related to the ions' absorption spectrum modulated by the photoisomerization probability. The approach is demonstrated using the polymethine dye HITC (1,3,3,1',3',3'-hexamethylindotricarbocyanine). The data show that both trans and cis forms of HITC(+) exist in the gas phase with trans→cis photoisomerization predominating over the 550-710 nm range and cis→trans photoisomerization occurring over the 735-770 nm range. The gas-phase photoisomerization action spectrum is comparable to the absorption spectra of trans HITC and cis HTIC in the condensed phase, but with the absorption peaks shifted to shorter wavelength. The gas-phase photoisomerization action spectrum of the (HITC)2(2+) dication dimer is also reported. (HITC)2(2+) cations photoisomerize over the 550-770 nm range to form more compact structures.