INTRODUCTION:
Imagine yourself at the
opening of an art exhibit at a museum: a special showing that you need
to have a ticket for. You happen to find yourself in a group with
art critics, tourists, and "normal" folks like yourself. As is
often
the case, the paintings are hung along a corridor, and uniformed guards
keep the flow of human traffic going "one-way". At the specified time,
the velvet rope is dropped, and you and your fellow museum goers enter
the corridor.
The tourists have less interest in the actual art than in being able to say they'd been to the museum when they get back home. And you can hear mutterings of "My 5 year old can draw better" and "Let's go see the monuments next". Unsurprisingly the tourists all arrive at the end of the exhibit quickly, in a tight little group.
Meanwhile, you and the rest of the general population are proceeding at a moderate pace, occasionally stopping to savor a particular oeuvre. You arrive at the end of the exhibit decidedly later than the tourists (who are probably back on their bus).
The critics however are
getting
rowdy. There is disagreement over the artist's brushstrokes, use
of pigment, and career developement. "He was a has-been by this
period".
"Who are you calling a 'has-been', you hack?" A fight breaks
out.
More guards are summoned. As you leave the building, you can
still
hear the rumpus. Needless to say, the critics one by one straggle
to the end of the exhibit last (doubtless greeted by a waiting police
van.)
Yes, this is still the CHM 245 web site, and what you just read was an analogy for chromatography, a technique of separating components of a mixture based on their relative affinities for a solid medium.
The people represent a
mixture
containing three components. Each component has a different
affinity
for the "stationary phase". In my story above that would be the
paintings
on the "solid support" (the walls). In liquid, or thin layer
chromatography,
the solid is silica or alumina (essentially sand or clay). In
gas/liquid
chromatography, the stationary phase might be a wax or thick oil that
coats
the solids support, perhaps crushed brick. All of that is packed
into a coiled copper tube called "the column".
The security guards are
the "mobile phase", keeping the flow of patrons going in one
direction.
In gas chromatography, this is the flow of inert
gas
through the instrument. In thin layer, or paper chromatography,
it's
capillary action, causing a liquid to rise up the stationary
phase.
In column chromatography, gravity keeps the liquid flowing down the
column.
The essence then of a
chromatographic
separation is the partitioning of a component between the mobile and
stationary
phases. If the sample likes to stick to the solid phase, it will
take a long time to reach the end of the column (or for PC and TLC, the
"spot" won't move very far). Hopefully, we have chosen our phases
so that the components of the mixture have different affinities - i.e.,
they don't all stick at the same location and not move, nor do they all
fly off to the end of the column at the same speed.
QUANTITATIVE ANALYSIS:
Gas chromatography
offers
a unique opportunity to determine the relative amounts of each mixture
component, while keeping sample size down to microgram amounts.
Other
techniques allow quantitation, but much more sample is required.
The output of the
GC is typically a graphed series of peaks. The relative areas
of the peaks are proportional to the relative amounts of the compounds
in the mixture. The area can be obtained by:
1) Triangulation - the
peak
is approximated by a triangle. The height and base are measured,
and the area is calculated.
2) Cutting and
weighing.
The original is photocopied several times, cut out with scissors, then
weighed on a balance. The relative weights are proportional to
the
areas.
3) Counting boxes.
If the output is on graph paper, the area can be obtained very
accurately
by counting the boxes under the curve.
4) An Integrating
Recorder can be used. This piece of equipment digitizes the
voltage
frome the GC, and calculates the relative areas of each peak, and
prints
the results. Yes, we have one at Alexandria.
INTERPRETATION OF RESULTS:
TWO PEAKS IN THE GC: The integrating recorder will give you percentages for each peak in your output. Ideally, only two peaks will appear for your fractions, one for ethyl acetate and one for butyl acetate. For now, let's assume that is the case. Because the thermal conductivity detector response increases for compounds with larger molecular weights and/or increasing polarity, a correction factor must be applied (Mf in your Gilbert book). Notice that the higher molecular weight butyl acetate has a lower correction factor than ethyl acetate. After application of the correction factors though, the resulting percentages need to be renormalized to 100%
Example: Let's
assume
that the recorder claims that the mixture is 50% ethyl and 50%
butyl.
Apllication of the correction factors gives (50)(0.89) = 44.5% for
ethyl,
and (50)(0.74) = 37% for butyl. Clearly a mixture of only ethyl
and
butyl acetates can't be 44.5% one and 37% the other. What's the
other
18.5 % ?!?
So we normalize the values
to add to 100%:
44.5/(44.5 + 37) = 54.6% and 37/(44.5 + 37) = 45.4%
Notice that proper application of the correction factors should lead to a lowering of the percentage of butyl acetate.
EXTRA PEAKS IN THE
GC:
Extra peaks might be due to air, acetone, water, or other
impurities.
In this case, because the refractive index analysis assumes only two
components
in the mixture, we must identify and normalize the two peaks due to our
acetates first, before applying the correction factors.
We
have to normalize again after the correction factors too, just like
before.
We need to do this because
the percentages derived from refractometry must add up to
100%.
Impurities are not (and cannot be) considered. For multiple
peaks,
no valid comparison can be made between refractometry and
chromatography
without the normalize-correct-normalize process.