SIMPLE AND FRACTIONAL DISTILLATION

Adriel T. David, Daryl M. David, Kimberly S. Delos Santos,

Francesca B. Echavez and Ruzelle A. Eroles
Group 3 2F Medical Technology Biochemistry Laboratory

ABSTRACT
There were mainly two things done in this experiment; preparation of buffer solution which in this case was Acetate
buffer, and electrometric and calorimetric determination of pH. Acetate buffer was prepared using 1.43mL of
CH3COOH and 0.1481g of NaOH pellets. Getting a ph of 4.03, with the aid of electric pH meter it was confirmed that
the buffer produced by the group conformed with the desired ph assigned which was ph 4.00. To determine the ph of
the juice, assigned sample to the group and the ph of the water, the buffer solutions were each tested calorimetrically
with the reagents thymol blue, bromophenol blue, bromocresol green, bromocresol purple, phenol red, methyl red,
methyl orange, and phenolphthalein. Observing the changes in each buffer solution, it was found out that the assigned
sample has a ph of 4.00 and water with a ph of 7.50.

INTRODUCTION
One of the more important properties of an
aqueous solution is its concentration of hydrogen
ion. The H+ or H3O + ion has great effect on the
solubility of many inorganic and organic species,
on the nature of complex metallic cations found
in solutions, and on the rates of many chemical
reactions. It is important that we know how to
measure the concentration of hydrogen ion and
understand its effect on solution properties.
For convenience the concentration of H+ ion is
frequently expressed as the pH of the solution
rather than as molarity. The pH of a solution is
defined by the following equation:
pH = -log[H+ ]
where the logarithm is taken to the base 10. If
[H+ ] is 1 x 10-4 moles per liter, the pH of the
solution is, by the equation, 4. If the [H +] is 5 x
10-2 M, the pH is 1.3.
We measure the pH of a solution experimentally
in two ways. In the first of these we use a
chemical called an indicator, which is sensitive to
pH. These substances have colors that change
over a relatively short pH range (about two pH
units) and can, when properly chosen, be used to
determine roughly the pH of a solution. Two very
common indicators are litmus, usually used on
paper, and phenolphthalein, the most common
indicator in acid-base titrations. Litmus changes
from red to blue as the pH of a solution goes
from about 6 to about 8. Phenolphthalein
changes from colorless to red as the pH goes
from 8 to 10. A given indicator is useful for
determining pH only in the region in which it
changes color. Indicators are available for
measurement of pH in all the important ranges of
acidity and basicity. By matching the color of a
suitable indicator in a solution of known pH with
that in an unknown solution, one can determine

the pH of the unknown to within about 0.3 pH

units.
Some solutions, called buffers, are remarkably
resistant to pH changes. Water is not a buffer,
since its pH is very sensitive to addition of any
acidic or basic species. Even bubbling your breath
through a straw into distilled water can change
its pH by at least 1 unit, just due to the small
amount of CO2 in exhaled air. With a good buffer
solution, you could blow your exhaled air into it
for half an hour and not change the pH
appreciably. All living systems contain buffer
solutions, since stability of pH is essential for the
occurrence of many of the biochemical reactions
that go on to maintain the living organism. There
is nothing mysterious about what one needs to
make a buffer. All that is required is a solution
containing a weak acid and its conjugate base.
An example of such a solution is one containing
the weak acid HB, and the B- ion, its conjugate
base, obtained by dissolving the salt NaB in
water. The pH of such a buffer is established by
the relative concentrations of HB and B- in the
solution. In such a solution [H+] can be calculated
by:

[H+] = Ka x

[HB ]

EXPERIMENTAL
A. Compounds tested (or Sample used)
Distilled water, Glacial acetic acid (99.7% w/w),
NaOH pellets, buffers at pHs 2.00, 3.00, 4.00,
5.00, 7.00, 7.50, 8.00, 9.00, 11.00, and 12.00,
and
Acid-base
indicators
(Thymol
blue,
Bromophenol
blue,
Bromocresol
green,
Bromocresol purple, Phenol red, Methy red,
Methyl orange, and Phenolphthalein) were used.

Juice sample was also used in the experiment in

the determination of its pH.

B. Procedure
1. Preparation of buffers
Two hundred fifty milliliters of 0.10M acetate
buffer pH 4.00 was prepared by the group.
Henderson-Hasselbach equation was used to
calculate the amounts of the buffer components
glacial acetic acid and NaOH pellets.
Computations:
Ratio of acid to conjugate base:
pH = pKa + log [A-]/[HA]
[A-]/[HA] = 10(4-4.76)
[A-] = 0.1738
[HA] = 1
Total theoretical moles of buffer:
= 1.1738
Total actual moles of buffer:
= (0.1M)(0.25L) = 0.025n
Let x be:
x + 0.1738x = 0.025n
x = 2.13 x 10-2n of CH3COO- from acid
0.1738x = 3.70 x 10-3n of CH3COO from
conjugate base

2.

Calorimetric determination of pH
a. Preparation of color standards using
the buffer solutions
Ten test tubes were labeled with pHs 2.00,
3.00, 4.00, 5.00, 7.00, 7.50, 8.00, 9.00,
11.00, and 12.00. One milliliter of a buffer of a
certain pH was pipetted into the test tube and
four milliliters of distilled water were added.
Eight sets of ten test tubes were prepared and
corresponded to a specific acid-base indicator.
Two drops of Thymol blue, Bromophenol blue,
Bromocresol
green,
Bromocresol
purple,
Phenol red, Methyl red, Methyl orange, and
Phenolpthalein were added respectively in
each of the set. Colors of the indicators were
observed and noted.
a. Determination of the pH of samples
Two sets of eight test tubes were prepared
and filled with 5ml of each sample. The first
set was filled with distilled water and the
second set was fruit juice. Two drops of the
eight acid-base indicators were used in each
set and each test tube of their specific
indicator were compared to the color
standards prepared using the buffer solutions.
The pHs of the samples were obtained by
observing where the color is nearest to.

Volume of glacial acetic acid:

99.7% = 99.7g/100g
Moles = 99.7g/(60g/n) = 1.66n
Volume = 100g/(1.05g/mL) = 95.24mL =
0.09524L
M = 1.66n/0.09524 = 17.43M
Volume CH3COOH = 0.025n/(17.43n/L) = 1.43
x 10-3 L = 1.43mL
Mass of NaOH:
m = 3.70 x 10-3n x (40g/n) = 0.1481g
1.43 mL of glacial acetic acid was measured
using serological pipette and 0.1481g of NaOH
pellets was measured using analytical balance.
These two compounds were mixed and
transferred together into a beaker. After
completely
dissolving
NaOH
pellets,
the
compounds were transferred to a 250.00 mL
volumetric flask. The rinsing of the beaker were
put into the volumetric flask which was then filled
with water up to its mark. The flask was covered
by parafilm and was shaken. After checking the
pH with the pH strip, the buffer solution was then
transferred into a clear reagent bottle which was
properly labeled.

Figure 1. Setup of buffer solutions

RESULTS AND DISCUSSION

With a pH of 4.00 assigned, the group was able
to produce a buffer solution with that met its
requirements. To further confirm the pH of the
solution, pH strip test and pH meter test was
applied. The pH strip test was found to be
positive and met the pH 4.00 requirement, while
the pH meter test also yielded a result of pH
4.03.

Using the different pH buffer solutions that

each group produced, we tested the efficiency of
the indicators which gave the following results:

Acid Base
Indicator

2.00

3.00

4.00

5.00

Yellow

Yellow

Yellow

Yellow

Table 2. PH range of different indicators

Acid Base Indicator

Thymol
Bromophenol
Blue

Yellow

Yellow

Light
Green

Violet

Bromophenol
Green

Light
Yellow

Light
Yellow

Light
Yellow

Light
Yellow

Bromocresol
Purple

Yellow

Yellow

Yellow

Yellow

Phenol Red

Yellow

Yellow

Yellow

Yellow

Methyl Red

Pink

Pink

Pink

Pink

Methyl
Orange

Orange

Light
Pink

Yellow

Yellow

Pehnolphthal
ein

Colorless

Colorless

Colorless

Colorless

Table 1. Acid-Base indicators

The diluted buffer solution for each pH was

introduced to the indicators which produced
different colors. These colors act as a guide for us
to identify the indicator used and the pH of our
substance.
The table tells us the different capacity of the
indicators when reacted to a series of varying pH
of buffer solutions. At a certain pH an indicator
changes its color, which helps us identify the
acidity or basicity of a certain substance. For this
experiment, we are trying to find range in which
an indicator changes its color, called endpoint.
Using the data that we got, we found the
endpoint for each indicator:

pH Range

Thymol

8.00 9.00

Bromophenol Blue

3.00 5.00

Bromophenol Green

5.00 7.50

Bromocresol Purple

5.00 7.50

Phenol Red

7.00 8.00

Methyl Red

5.00 7.50

Methyl Orange

2.00 4.00

Pehnolphthalein

7.50 9.00

Using the pH range of our indicators we can

now identify the pH of our sample liquid, by
comparing the results.
Table 3. Analysis of distilled water and sample
liquid

Acid Base
Indicator

Sample
Distilled
Water

Juice
Sample

Yellow

Light Yellow

Blue Violet

Light Green

Light Blue

Light Yellow

Light Yellow
Green

Yellow

Phenol Red

Yellow

Yellow

Methyl Red

Light Orange

Light Pink

Thymol
Bromophenol
Blue
Bromophenol
Green
Bromocresol
Purple

Methyl Orange

Orange

Baby Pink

Pehnolphthalein

Colorless

Colorless

pH

pH 7.50

pH 4.00

Based on the results, we can now conclude that

the pH of our distilled water is 7.50 and the pH of
our sample liquid (juice) is 4.00.

REFERENCES
[1] pH Measurements - Buffers and Their
Properties. Dr. Cielito DeRamos King. Retrieved
from http://webhost.bridgew.edu/c2king/CHEM
142/Lab/7_Buffers%20and%20Properties.pdf
(02/16/15)