Chapter 17: Fundamentals of Spectrophotometry

absorbance

- (A) common logarithm of the reciprocal of the transmittance of a pure solvent
- also called optical density
- directly proportional to the concentration

absorption spectrum

- graph showing how (A) varies with wavelength
- absorption has higher energy than fluorescence and phosphorescence

Beer's Law

- the heart of spectrophotometry of applied analytical chemistry
- The mathematical relationship between concentration and absorbance
- fails in concentrated solutions

chemiluminescence

- emission of light from a chemical reaction
- ex light from a firefly or light stick

chromophore

the part of the molecule responsible for light absorption

cuvet

- a cell that contains the liquid sample for visable end ultraviolet spectroscopy
- has flat, fused-silica (SiO?) faces or glass
- 1.000 cm path length

derivatization

the chemical alteration of an analyte so that it can be detected or separated easily from other species

electromagnetic spectrum

All of the frequencies or wavelengths of electromagnetic radiation

electronic transition

- an electron from one molecular orbital moves to another orbital, with a constant increase or decrease in the energy of the molecule
- usually involve vibrational and rotational transitions
** electronic absorption bands are usually broad because many di

emission spectrum

- graph of emission intensity vs emission wavelength
- constant ?ex and variable ?em
- measure emitted radiation rather than fraction of incident radiation striking the detector

excitation spectrum

- measure by varying the excitation wavelength and measuring emitted light at one particular wavelength (?cm)
- graph of emission intensity vs excitation wavelength; look slike absorbtion spectrum
- constant ?em and variable ?ex

excited state

when a molecule absorbs a photon and the energy of the molecules increases

fluorescence

- emission of a photon during a transition between states with the same spin quantum numbers
- radiational transition S?? S?
- relatively rare
- lifetime: 10?? to 10??s

frequency

(1/second) number of complete oscillations that a wave makes each second

ground state

the lowest energy state of a molecule

hertz

one oscillation per second

irradiance

- (P) the energy/time unit area in the light beam
- watts per square meter (W/m�)
- also called intensity or radiant power

luminescence

- emission of light from an excited molecule
- generally more sensitive than absorption - sensitive enough to observe single molecules

masking

- when reagens like neocuprine or thiourea form strong complexes that prevent Cu�? from reacting with ferrozine

molar absorptivity

- characteristic of a substance that tells how much light is absorbed in a particular wavelength
- also called extinction coefficient
- units: M?� cm?�

molecular orbital

describe the distribution of electrons in a molecule

monochromatic light

- consists of 1 color (one wavelength)
- the better the ______________, the narrower the range of wavelengths in the emerging beam

monochromator

- what light is passed through to select one wavelength
- ex. prism, grating or filter

phosphorescence

- emission of a photon during transition between states with different spin quantum numbers
- radiational transition T?? S?
- has lower energy than fluorescence; energy comes at longer wavelengths
- relatively rare
- lifetime: 10?? to 10�s; involves chang

photon

a particle of electromagnetic radiation having zero mass and carrying a quantum of energy

reagent blank

- contains all reagents but with analyte replaced by distilled water
- important in most spectrophotometric analyses
- must be subtracted from the absorbance of samples and standards before doing any calculations

refractive index

(n), a measure of the light-bending ability of a medium
- for visible wavelengths, n>1
- when light moves between media with different refractive indexes, frequency remains constant but wavelength changes

rotational transition

changes in the rotational states

self-absorbtion

- absorption of excitation or emission energy by analyte molecules in the solution
- also called the inner filter effect

singlet state

- the state in which the spins are opposed
- S?= lowest energy state
- not split by a magnetic field

spectrophotometry

- any technique that uses light to measure chemical concentrations

transmittance

- (T) the fraction of the original light that passes through the sample
- ranges from 0-1

triplet state

- if the spins are parallel
- T?= lowest energy state; T?< S?
- split into 3 slightly different energy levels in a magnetic field

vibrational transition

- change in the vibrational state
- usually involve simultaneous rotational transistions

wavelength

(?) crest to crest distance between waves

wavenumber

(v=1/?)
The reciprocal of wavelength; the number of waves in a unit length or distance per cycle

nonbonding orbital

(n) highest energy occupied orbital
- ex. formaldehyde: mostly O

antibonding orbital

(pi*) lowest energy unoccupied orbital
- produce repulsion, rather than attraction for the C & O atoms

vibrational relaxation

- usually the first process after absorbtion
- relaxation to the lowest vibrational level of S1

radiationless transition

- vibrational energy is transferred to other molecules through collisions, not by emission of a photon
- net effect: convert part of energy of absorbed photon into heat throughout medium

internal conversion

- when a molecule enters a highly excited vibrational level of So, having the same energy as S1
- radiationless transition between states with the same spin quantum numbers (S?? S?)

intersystem crossing

- radiationless transition between states with different spin quantum numbers (T?? S?)

vibrational energy

- observed in non-polar or less polar solvents
- broadened beyond recognition in polar solvents

solution phase spectra

- broadened absorption spectrum
- absorbing molecules are surrounded by solvent molecules with a variety of orientations that create slightly different energy levels for different absorbing molecules

simple gas phase molucules

not in close contact with neightbos, limited # of energy levels
- have extrememly sharp absorptions

essentials of a luminescence experiment

- sample is irradiated at 1 wavelength
- emission is observed over a range of wavelengths
- excitation monochromator selects teh excitation wavelength
- emission monochromator selects one wavelength at a time to obserce