SHELF LIFE DETERMINATION

Santos, G.
1
, Silverio, A.
1
, Soriano, R.
1
, Tan, J.
1
, Tequillo, H.
1

University of Santo Tomas, Faculty of Pharmacy
1
Group 8 3F Pharmacy Physical Pharmacy Laboratory

ABSTRACT
The experiment was performed to understand the process of shelf life determination. Acetylsalicylic acid, also
known as aspirin, is one of the most commonly used analgesics worldwide and was used for this experiment. From a
stock solution of salicylic acid, five standard solutions were prepared and their absorbance values were determined
using the UV-VIS spectrophotometer at 296 nm. The best fit line was determined using linear regression. A sample
solution containing aspirin dissolved in water was placed in an oven at 50C. The absorbance readings at 296 nm
were taken after 15, 30, 45, 60 and 90 minutes. From this data, the concentrations of aspirin remaining together with
its logarithmic equivalent were determined. Using linear regression, the best fit line was once again obtained, which
is essential for the computation of shelf-life.
INTRODUCTION
Shelf life is best defined as the time span
over which the quality of a product remains within
specifications. That is, it is the time period over
which the efficacy, safety, and esthetics of a product
can be assured. When the degradation of the essential
components cannot be adequately described by a rate
expression, shelf life cannot be easily estimated or
projected.
Estimation of product shelf-life is done by
two methods estimation from data obtained under
the same conditions as those that the final product is
expected to withstand and the estimation from tests
conducted under accelerated conditions. [4]
Half-life is the amount of time required for a
quantity to fall to half its value as measured at the
beginning of the time period. It is used to describe a
quantity undergoing exponential decay, and is
constant over the lifetime of the decaying quantity.
[8]
Aspirin, also known as acetylsalicylic acid,
is a derivative of salicylic acid. It is the most widely
used medicine in the world. It has the ability to
reduce fever (antipyretic), reduce pain (analgesic),
relieve swelling and pain associated with rheumatism
and arthritis (anti-inflammatory), enhance the
elimination of uric acid (uricosuric), and interfere
with the bodys production of prostaglandins and
their products (thromboxanes), which are associated
with headaches, platelet aggregation and
vasoconstriction. [1]


Aspirin is a part of a group of medications
called nonsteroidal anti-inflammatory drugs
(NSAIDs), but differs from most other NSAIDs in
the mechanism of action. Aspirins ability to suppress
the production of prostaglandins and thromboxanes is
due to its irreversible inactivation of the
cyclooxygenase (COX) enzyme. Aspirin acts as an
acetylating agent where an acetyl group is covalently
attached to a serine residue in the active site of the
COX enzyme. This makes aspirin different from
other NSAIDs (such as diclofenac and ibuprofen),
which are reversible inhibitors. [3]

Aspirin undergoes hydrolysis with the
resultant degradation products being salicylic acid
and acetic acid. [2]

Figure 1. Hydrolysis of Aspirin
Ultraviolet-visible spectrophotometry (UV-
Vis) is the absorption spectroscopy or reflectance
spectroscopy in the ultraviolet-visible spectral region.
This means it uses light in the visible and adjacent
(near-UV and near-infrared) ranges. The absorption
or reflectance in the visible range directly affects the
perceived color of the chemicals involved. Beer-
Lambert law is most often used to determine
concentrations quantitatively. The instrument used is
UV-Vis spectrophotometer. [6]
The Arrhenius equation is a simple but
remarkably accurate formula for the temperature
dependence of reaction rates. The equation was
proposed by Svante Arrhenius in 1889, based on the
work of Dutch chemist Jacobus Henricus vant Hoff
who had noted in 1884 that Vant Hoffs equation for
the temperature dependence of equilibrium constants
suggests such a formula for the rates of both forward
and reverse reactions. Arrhenius provided a physical
justification and interpretation for the formula.
Currently, it is best seen as an empirical relationship
and can be used to model the temperature variation of
diffusion coefficients, population of crystal
vacancies, creep rates, and many other thermally-
induced processes or reactions. [7]
MATERIALS AND METHODS
A. Compounds Tested
Aspirin has the chemical formula C
9
H
8
O
4
and the
IUPAC name 2-(acetyloxy) benzoic acid. It has the
molecular weight of 180.16g/mol. It has a density of
1.40g/cm
3
and a melting point of 136
o
C. [5]

B. Procedure
1. Preparation of Standard Solutions
20 mg of salicylic acid was weighed accurately and
was transferred into a 100mL volumetric flask. The
volume was filled with distilled water. (Stock
solution) 20mL from stock solution was transferred
to a 50mL volumetric flask and was diluted to
volume. It was labeled as Solution A. Another 20mL
from the stock solution to a 100mL volumetric flask
and, again, was diluted to volume. It was labeled as
Solution B. Next, 20mL from Solution A was
transferred to a 50mL volumetric flask and was
diluted to volume. It was labeled as Solution C. 20mL
from Solution A was transferred to a 100mL
volumetric flask and was diluted to volume. It was
labeled Solution D. 20 mL from Solution C was
transferred to a 50mL volumetric flask and was
diluted. It was labeled as Solution E.
2. Preparation of the Standard Absorbance
Concentration Curve
Absorbance reading of the prepared standard
solutions were determined at 296nm using the UV-
vis spectrophotometer. The absorbance values were
plotted against the known Salicylic acid
concentration. Linear regression was used to obtain
the best fit line.
3. Preparation of Sample Solutions
40mg of Aspirin was weighed and transferred into a
500mL volumetric flask. Volume was filled using
distilled water. Enough samples were taken and
subjected to absorbance reading at 296nm. 100mL of
the solution was transferred into 4 separate beakers
and was placed in ovens set at 50C.
4. Preparation of Sample Solutions
Enough samples were gotten at 15, 30, 45, 60, 90,
and 120 minutes. The absorbance reading for each
sample was taken at 296nm. The concentration of
Aspirin remaining was determined based on the
standard absorbance-concentration curve. Log C of
the Aspirin vs. the remaining time was plotted. Then,
the degradation rate constant (k) of Aspirin was
determined for each temperature setting. Log k vs.
1/T was plotted. Using the Arrhenius equation, the
degradation rate constant at 25C was calculated.
RESULTS AND DISCUSSION

The experiment was conducted with a barometric
pressure of 74.7cmHg and a room temperature of
25.6
o
C.

Upon calculation of the concentration of each
solution prepared, the following values were obtained
(Table 1). Absorbance readings of each of the
prepared solution were done at 296nm using the UV-
VIS spectrophotometer and results can be seen
below.
Computation for the concentration of each solution
follows the formula C
1
V
1
=C
2
V
2
as seen below.
Stock Solution = 0.20mg/mL
Solution A
C
1
V
1
=C
2
V
2
0.20mg/mL (20mL) =C
2
(50mL)
C
2
=0.08mg/mL
Solution B
C
1
V
1
=C
2
V
2
0.20mg/mL (20mL) =C
2
(100mL)
C
2
=0.04mg/mL
Solution C
C
1
V
1
=C
2
V
2
0.08mg/mL (20mL) =C
2
(50mL)
C
2
=0.032mg/mL
Solution D
C
1
V
1
=C
2
V
2
0.08mg/mL (20mL) =C
2
(100mL)
C
2
=0.016mg/mL
Solution E
C
1
V
1
=C
2
V
2
0.032mg/mL (20mL) =C
2
(50mL)
C
2
=0.013mg/mL
The a and b values were obtained after performing
linear regression in a calculator using the stat
function.

In graph form, here are the results.

Graph 1. Standard concentration absorbance curve

The concentration of aspirin solution was obtained by
computation. The absorbance reading was 1.167.
40mg/500mL = 0.08mg/mL

In order to obtain the concentration of salicylic acid
remaining as time passes at a temperature of 50
o
C,
the linear regression equation was utilized. The
tabulated results can be seen in Table 2.

y = bx + a
Where y = absorbance of sample
b = best fit line from standard solutions
a = a value from standard solutions
x = concentration of salicylic acid remaining
15 mins
y = bx + a
0.986 = 14.32x + 1.06
x = -5.447X10
-3
mg
30 mins
y = bx + a
1.002 = 14.32x + 1.06
x = -4.3297X10
-3
mg
45 mins
y = bx + a
1.022 = 14.32x + 1.06
x = -2.933X10
-3
mg
60 mins
y = bx + a
1.025 = 14.32x + 1.06
x = -2.723X10
-3
mg
90 mins
y = bx + a
1.030 = 14.32x + 1.06
x = -2.374X10
-3
mg


The concentration of acetylsalicylic acid remaining
as time passes at 50
o
C is taken from the computed
concentration of aspirin and the computed
concentration of salicylic acid, using the formula:
conc. of ASA = original conc. ASA SA remaining

15 mins
conc. of ASA = original conc. ASA SA remaining
x = 0.08 - (-5.447X10
-3
)
x = 0.085447 mg
30 mins
conc. of ASA = original conc. ASA SA remaining
x = 0.08 (-4.3297X10
-3
)
x = 0.0843297 mg
45 mins
conc. of ASA = original conc. ASA SA remaining
x = 0.08 (-2.933X10
-3
)
x = 0.082933 mg
60 mins
conc. of ASA = original conc. ASA SA remaining
x = 0.08 (-2.723X10
-3
)
x = 0.082723 mg
90 mins
conc. of ASA = original conc. ASA SA remaining
x = 0.08 (-2.374X10
-3
)
x = 0.082374 mg

The logarithmic equivalent of each value is taken in
order to plot the data in a graph. Using the formula:
x = log (conc. of ASA)

15 mins
x = log (conc. of ASA)
x = log (0.085447)
x = -1.0683
30 mins
x = log (conc. of ASA)
x = log (0.0843297)
x = -1.0740
45 mins
x = log (conc. of ASA)
x = log (0.082933)
x = -1.0813
60 mins
x = log (conc. of ASA)
x = log (0.082723)
x = -1.0824
90 mins
x = log (conc. of ASA)
x = log (0.082374)
x = -1.0842


The a and b values were obtained after performing
linear regression in a calculator using the stat
function.


Graph 2. Log concentration of aspirin vs. time graph

Using Arrhenius equation, the degradation rate
constant at 25
o
C was found as well as the half-life of
the drug and its shelf life.

Where k2 = degradation rate constant at T2
k1 = degradation rate constant at T1
Ea = activation energy
R = gas constant
T1 = first temperature
T2 = second temperature

The degradation rate constant at 25
o
C is
3.35x10
-5
/min.

For the computation of half-life, the following
formula is used:

The half-life of the drug is at 14 days, 8 hours and 47
minutes.

In the computation of shelf-life, the following
formula is used:

The shelf life of the drug is at 2 days, 4 hours and 14
minutes.




CONCLUSION
Shelf-life is the time required for 10% of the drug to
degrade and 90% of the drug to remain intact. The
computed value for aspirin at 25
o
C is 2 days, 4 hours
and 14 minutes. Half-life is the time required for 50%
of the drug to degrade and 50% of the drug to remain
intact. The computed value for aspirin at 25
o
C is 14
days, 8 hours and 47 minutes.

REFERENCES
[1] Bayquen, A., et. al. (2009). Synthesis of
Acetylsalicylic Acid (Aspirin). In Laboratory
Manual in Organic Chemistry (p. 141). Quezon City,
Philippines: C & E Publishing.
[2] https://pharmahub.org/resources/535/download/
ASA_Freshman_Lab_Handout.pdf
[3] http://en.wikipedia.org/wiki/Aspirin
[4] Yoshioka, S., et. Al (2000). Stability of Drugs and
Dosage Form. Springer Science and Business Media.
[5] http://en.wikipedia.org/wiki/Aspirin
[6] http://en.wikipedia.org/wiki/Ultraviolet%E2%
80%93visible_spectroscopy
[7] http://en.wikipedia.org/wiki/Arrhenius_equation
[8] http://en.wikipedia.org/wiki/Half-life