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MCAT Prep > Analytic Techniques > Flashcards

Analytic Techniques Flashcards

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MCAT flashcards
1
Q

Extraction in protein purification

A

have to lyse cell membranes to remove proteins

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2
Q

What do you treat protein mixtures with during extraction?

A

protease inhibitors

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3
Q

Where are proteins in centrifugation?

A

lighter proteins stay suspended in the supernatant

heavier cell debris and such moves to the bottom

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4
Q

How can you use salt to separate proteins?

A

initially the salt will increase solubility of the protein by breaking apart side chain reactions

then, when enough salt is added it will compete with the protein for solvent to be broken down and the protein will precipitate out

use the optimal amount of salt that the protein precipitates out

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5
Q

Thin layer chromatography

A

use polar silica as stationary phase

separate compounds on polarity

use Rf ratio

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6
Q

What type of molecules move the furthest in thin layer chromatography?

A

nonpolar substances

interact poorly with the polar stationary phase

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7
Q

Rf

A

ratio of how far a substance traveled in TLC

has to be between 0-1

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8
Q

Iodine in TLC

A

helps us visualize the compounds

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9
Q

High performance liquid chromatography

A

uses a polar stationary phase

polar materials have a longer retention time

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10
Q

Reverse phase HPLC

A

uses a nonpolar stationary phase

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11
Q

Gas chromatography

A

mobile phase is gas and stationary phase is liquid

compounds with low boiling points evaporate first

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12
Q

Size exclusion chromatography

A

captures small molecules in gel

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13
Q

Anion-exchange chromatography

A

has positive beads to attract anions

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14
Q

Cation-exchange chromatography

A

has negative beads to attract cations

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15
Q

Salt buffer in ion-exchange chromatography

A

releases attracted molecules

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16
Q

Affinity chromatography

A

ligand binds to a specific receptor in the column

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17
Q

Electrophoresis anode and cathode

A

anode is positive

cathode is negative

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18
Q

SDS limits

A

does not break down disulfide bridges

need a reducing agent to break down disulfide bridges

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19
Q

What travels the furthest in SDS page?

A

smaller molecules

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20
Q

Isoelectric focusing

A

a type of electrophoresis where molecules will end up at pH that equals there pI

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21
Q

Immunoassays

A

rely on antibodies for specificity/identification

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22
Q

Example of an immunoassays

A

Western blotting

23
Q

ELISA

A

antigens marked with fluorescent labels bind the molecule of interest can use spectroscopy to see how many molecules binded

24
Q

Beer-Lambert Law

A

A = ecl

25
Q

At what wavelength do proteins have max absorbance? Why?

A

in the UV spectrum

as aromatic/conjugated pi bonds

26
Q

Extraction

A

separate liquid mixtures into organic and aqueous phases

27
Q

Organic phase in extraction

A

non-polar

28
Q

Aqueous phase in extraction

A

polar

29
Q

Where are uncharged acid and bases found in extraction?

A

in the non-polar organic phase

30
Q

Distillation

A

separates liquids by their boiling point

31
Q

Fractional distillation

A

can separate liquids with similar boiling points

32
Q

Vacuum distillation

A

lowers atmospheric pressure to allow high boiling points to evaporate

33
Q

Recrystallization

A

recrystallize a solid precipitate to filter out impurities that may have been in the original precipitate

34
Q

Nucleation

A

initiates the recrystallization process

35
Q

Spectroscopy

A

analyzes molecules based on interaction with electromagnetic radiation

36
Q

How does IR radiation work?

A

makes polar bonds bend and stretch

37
Q

Where is O-H found in IR?

A

a broad peak at 3100-3500

38
Q

Where is C-H found in IR?

A

a stretch at 3000

39
Q

Where is C=O found in IR?

A

a sharp peak at 1650-1780

40
Q

UV-Vis Spectroscopy

A

looks at movement of electrons

good for determining conjugated pi systems

41
Q

How can you find the color of a compound from UV-VIs spectroscopy?

A

colors that you don’t absorb give the color of the conjugated system

42
Q

What is UV-Vis spectroscopy good at measuring?

A

conjugated pi systems / aromatic compounds

43
Q

Where are less shielded protons found in NMR?

A

downfield to the right

44
Q

What makes a proton less shielded in NMR?

A

the presence of electronegative species that remove ED

45
Q

How many number of signals are found in NMR?

A

number of sets of equivalent hydrogens / carbons

46
Q

Where is the position of signals in NMR?

A

look at shielding to shift upstream or downstream

47
Q

What does the size of the signal in NMR tell us?

A

more protons means more area/larger size

48
Q

How is splitting in NMR determined?

A

n+1, where n is the number of adjacent hydrogens

49
Q

Is there splitting in 13C NMR?

A

no

50
Q

Mass spectroscopy steps

A

1) Ionize by shooting electrons at molecule
2) Accelerate in E-field
3) Curve in B-field to tell us the mass

51
Q

Parent ion in mass spectroscopy

A

Generated by the loss of only on electron

Is the peak the furthest to the right on graph

52
Q

What can the parent ion tell us in mass spectroscopy?

A

the molecular weight of the molecule

53
Q

Parent ion abbreviation

A

M+

MCAT Prep (22 decks)

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