Absorption Spec Flashcards
What is a chromophore
A group of bonded atoms (that are part of a molecule) and act as a unit for excitations to that they have a characteristic spectrum
What are UV visible chromophores
Give examples of
Aromatic of conjugated systems that loose their energy difference into the uv range
Ex benzene, indole, phenol, amide, aromatic bases of nucleic acids
What are infrared chromophores
They have vibrations that involve the stretching or bending of bonds
What are examples of chromophores in both uv and IR
Benzene (UV)
Amide 1 vibration (IR)
Slide 3 max absorbances
Write down on sheet
How to find amount of amides in a protien
AA- 1-asn-gln
Why is the extinction coeffienct on the y axis of the absorption graph at log scale
To see the high extinction coefficients at low wavelengths
What is speacial about the number of absorption peaks for a single chromophore
Many diff absorption peaks at diff wavelength
What is an example of why we need buffer when measuring absorbance
At ph 12 tyrosine gets deprotonated and turns to tyrosinate
This changes it’s absorbance from the ph 7 tyrosine, which is why we need a buffer to keep a certain ph
Does the uv spectrum of trp change at high ph
No
Under diff ph conditions, why would amides in the peptide back bone absorb diff
Because the polypeptide shape is diff at diff ph
Can form helix, random coil, or beta strands
For lysine at What pH is each formed
helix,
random coil,
beta strands
helix, pH 10.8 low temp (25 deg)
random coil, 6 (25 deg)
beta strands 10.8, high temp (52 deg)
Why would lysine be random coil at ph 6?
It’s pka is 10.8 so at ph 6 it’s protonated
Carrying a charge make it change conformation
The spectrum of a molecule is ____
The sum of the spectra for all chromophores present
Like in a graph it’s show a lot of peaks because that’s all the chormophores
Where do nucleic acids absorb
Higher energy transitions
Why do we not measure spectra below 190nm
260nm
Idk
Because O2 absorbs there and we don’t want that measurement