Characterizing biological macromolecules with attenuated total reflectance-Fourier transform infrared spectroscopy provides hands-on spectroscopy experiences for undergraduates.

Infrared (IR) spectra of biologically derived materials display distinct absorption bands correlating to individual macromolecules: protein, polysaccharide, lipid, and nucleic acids. A series of experiments aimed at teaching qualitative bioorganic spectroscopy using attenuated total reflectance (ATR) Fourier transform infrared (IR) with biological polymers as samples is proposed. Labs targeting 1st and 4th year undergraduate students at St. Francis College are being developed. During 2014 âž” 2017, an integrated biology/chemistry exercise featuring elementary spectroscopy as an addition to an existing lab on light microscopy was administered to three sections of a 1st year general biology course. Students were taught the concept of a spectral fingerprint and to identify carbohydrate and protein based materials by looking for key vibrational bands. The success of that effort as determined by the results of an assessment quiz became the motivation for developing an advanced 4th year exercise involving four macromolecules. In a trial lab (Spring 2019) students gathered reference spectra from materials homogeneous in a single biopolymer followed by spectra of whole tissues which they were expected to fully characterize. Assessment data suggest that 1st year students benefited most from the experience. A detailed discussion of reference and sample spectra (as obtained by students) and relevant bond vibrations along with suggestions for instructors are presented.

Solvent effects on the resonance Raman and hyper-Raman spectra and first hyperpolarizability of N,N-dipropyl-p-nitroaniline.

Linear absorption spectra, resonance Raman spectra and excitation profiles, and two-photon-resonant hyper-Rayleigh and hyper-Raman scattering hyperpolarizability profiles are reported for the push-pull chromophore N,N-dipropyl-p-nitroaniline in seven solvents spanning a wide range of polarities. The absorption spectral maximum red shifts by about 2700 cm(-1), and the symmetric -NO2 stretch shifts to lower frequencies by about 11 cm(-1) from hexane to acetonitrile, indicative of significant solvent effects on both the ground and excited electronic states. The intensity patterns in the resonance Raman and hyper-Raman spectra are similar and show only a small solvent dependence except in acetonitrile, where both the Raman and hyper-Raman intensities are considerably reduced. Quantitative modeling of all four spectroscopic observables in all seven solvents reveals that the origin of this effect is an increased solvent-induced homogeneous broadening in acetonitrile. The linear absorption oscillator strength is nearly solvent-independent, and the peak resonant hyperpolarizability, beta(-2omega;omega,omega), varies by only about 15% across the wide range of solvents examined. These results suggest that the resonant two-photon absorption cross sections in this chromophore should exhibit only a weak solvent dependence.

Characterization of small unilamellar vesicles using solvatochromic pi* indicators and particle sizing.

A suite of small unilamellar vesicles (SUVs) composed of mixtures of phospholipids and cholesterol (CH) or the synthetic surfactant dihexadecyl phosphate (DHP) and cholesterol was investigated using a homologous series of solvatochromic pi* indicators coupled with size exclusion methods and photon correlation spectroscopy (PCS). The solvatochromic method, which is based on the measurement of solvent-dependent shifts in lambda(max) from UV-Vis spectra of solubilized indicators, was used to quantify the dipolarity and polarizability (pi*) of probe solvation environments. The partitioning of the series of individual di-n-alkyl-p-nitroaniline (DNAP) pi* indicators in PG(24)PC(46)Chol(30) and DHP(70)Chol(30) SUVs was examined as a function of the head group structure as well as the method of dye-vesicle preparation. Solubilization of the larger more hydrophobic probes in the bilayer portion of PG(24)PC(46)Chol(30) SUVs was aided through physical entrapment. Such physical methods were not needed for the smaller indicators or for the range of indicators in the DHP(70)Chol(30) dispersions. Extrusion and size-exclusion chromatographic methodologies for the preparation of physically entrapped dopants in SUVs of fixed size range demonstrated that the larger (longer alkyl chain) dopants in the series resided in PG(24)PC(46)Chol(30) liposomes with a wider range of sizes, while the smaller more polar solutes tended to be entrapped in smaller vesicles with a narrower size range.