Chromatographic Analysis of a Commercial Analgesics Active Component

I. Abstract
This study on Chromatographic Analysis of a Commercial Analgesics Active
Component seeks to separate, identify, quantify and analyze the active
component found in Biogesic, which may be useful for public awareness
despite its wide acceptance and availability in the market. Furthermore, this study
applies thin layer chromatography (TLC) technique since it is the simplest,
cheapest yet most effective approach in terms to the studys objectives. The TLC
method implies Rf (Rate of flow) values where it is given by the distance from
origin to center of subtance spot over the distance from origin to solvent front. As
a result, the TLC analysis of Biogesic yields the following Rf values where the
given distance from origin to solvent front measures 59 mm: 1.) acetaminophen
has a Rf value of 0.48; 2.) aspirin has a Rf value of 0.64; 3.) caffeine has a Rf
value of 0.22; 4.) Biogesic sample has a Rf value of 0.48 ; 5.) ibuprofen has a
Rf value of 0.73; and 6.) mefenamic Acid has a Rf value of 0.76. Thus, the study
shows that by having equal Rf values of about 0.48, Biogesics most active
component is acetaminophen (paracetamol), a relatively polar compound having
a molecular formula C8H9NO2(MW= 151.1626). [1]

II. Introduction
Analgesics (also referred to as painkillers) are any member of the group of drugs
used to relieve pain. The relief that analgesics bring in alleviating pain occurs
either by obstructing the pain signals that are to be transmitted by the peripheral
nervous systems receptors to the central nervous system or by interfering with
the interpretation of pain signals transmitted to the brains central nervous
system, without causing any loss of consciousness. However, it must be noted
that analgesics are distinct from anesthetics since they do not reversibly
eliminate sensation. [2]

Analgesics may be purely made of a single component or two (e.g. plain aspirin
or ibuprofen with mefenamic acid), or may be a combination of other several
components (e.g. acetaminophen together with mefenamic acid and ibuprofen).
Basically, components of analgesics are of two kinds, these are the narcotics and
the non-narcotics[3]. The narcotic agents (e.g. Ibuprofen) regularly recognized for
bringing adverse narcotic effects (e.g. drowsiness) can also be subdivided further
into two groups; the opiates (e.g. morphine, codeine and thebaine) which are
alkaloids of opium; and the opioids (e.g. oxycodone and hydrocodone) which is
any synthetic narcotic that resembles the naturally occurring opiates. In contrast
to the narcotic forms, the non-narcotic forms of analgesics are simply opposed to
the adverse effects that narcotic forms bring; acetaminophen, aspirin and
mefenamic acid are common compounds found in most analgesics of this kind.

Moreover, active components of an analgesic can be readily separated and
analyzed, or may be identifed and quantified, by employing easy and reliable
techniques. Perhaps, one of the best methods ever available to be performed in
most laboratories (after the first successful separation of pigments of leaf extracts
through a column packed with a chalk by Russian chemist Mikhail Tsweet) is thin
layer chromatography (TLC), a versatile technique used for the examination and
chemical separation of complex mixtures which is based upon the rates at which
the components of a mixture are carried through a stationary phase by a mobile
phase, also it basically involves the separation of mixtures due to differences in
the distribution coefficient(equilibrium distribution) of sample components
between two different phases, hence the simplicity and rapidity of this technique
allows it to be regularly used to monitor progress of organic reactions and to
check purity of products.

In view of the fact that thin layer chromatography deeply allows one aiming to
identify (Qualitative Analysis) and quantify (Quantitative Analysis), or maybe to
separate and analyze the active component of commercial analgesics, this
enables one to employ and test the efficiency of this technique to Biogesic, a
commonly used analgesic sold availably in the Philippines, without any
prescription required.

III. Data and Results
I. Chromatogram:

Ibuprofen Mefenamic acid
Aspirin
Acetaminophen Biogesic sample
Caffeine

**As viewed under UV light

II. Rf Determination:
Formula:
Distance from origin to center of substance spot
Rf value = Distance from origin to solvent front

**The table below shows corresponding Rf value at which each compound is
relative to the distance from the origin to its blots center:
Given: Distance from origin to solvent = 59 mm.
|Compound |Distance from origin to center of substance spot (in |Rf value |
| |mm) | |
|Acetaminophen |28.5 |0.48 |
|Aspirin |38 |0.64 |
|Caffeine |13 |0.22 |
|Sample (Biogesic) |28.5 |0.48 |
|Ibuprofen |43 |0.73 |
|Mefenamic Acid |45 |0.76 |

IV. Discussion
As the TLC plate was first loaded with the Biogesic sample that we wished to
be separated, it later developed by allowing the eluent to flow through the
adsorbant, in particular; by capillary action. We expected that the sample will
distribute itself either to the moving eluent or will stay in the adsorbents active
site. This is for the assumption that analytes having the most high interaction to
the eluent (mobile phase) will be moved due to attraction (capillary action), while
those analytes having the most high interaction to the adsorbent (mostly polar)
will remain settling at the TLC plate (stationary phase) itself. Also, at any given
rate there will be some sort of a competition and that the distribution rate can
differ from one compound to the other, as some components move faster with
the flowing solvent than those of the other components that might be stuck on the
adsorbant. Remember, the basis for this separation also depends on the
compounds structure (e.g. polarity and shape of the molecule). Hence, we can
readily assume from here that the stationary phase follows a strictly increasing
adsorptive power for polar molecules and that the mobile phase follows a strictly
increasing eluting power which ranks polar molecules from the TLC plates lower
portion up to the non-polar molecules.

In view of the concept behind TLC, one can say that the order of blots on the
chromatograms mobile phase, in accordance to their respective adsorbents, is
based according to how much the analyte is attracted to the eluent or simply
based accordingly to its increasing eluting power. Notice, the non-polar
compounds are situated at the TLC plates upper portion and that as other
compounds increases in polarity, it gets closer to the origin. Hence, we can
readily assume from here that the blots follow an increasing polarity opposite to
its order on the chromatogram. For instance, the chromatogram (as shown on
results) shows that the blot for caffeine, as situated closer to the origin, is more
distant from that of Acetaminophen or Biogesic sample used, thus one can say
from here that caffeine is more polar than Acetaminophen or the Biogesic
sample used, and that same assumption is true as we compare Acetaminophen
and Biogesic sample to Aspirin and so on. Consequently, the chromatogram
shows the following according to its increasing polarity (non-polar to polar) :
Mefenamic acid < Ibuprofen < Aspirin < Acetaminophen and Biogesic sample <
Caffeine. Furthermore, same assumption can be drawn from the computed Rf
values (as shown on the table), where the compound having the highest Rf value
is the most non-polar and the lowest as the most polar.

Nevertheless, one of the most important and critical aim of this study is the
identification of the most active component in the Biogesic sample used. It was
found out that both acetaminophen and Biogesic yield equal Rf values of about
0.48. Therefore, we can readily assume that the active component found in
Biogesic is acetaminophen (paracetamol).
V. Experimental
Analgesic sample was prepared. Biogesic tablet (500 g) was triturated with the
aid of mortar and pestle. The triturated Biogesic sample was mixed together
with methanol-toluene (2 mL, 1:1 mole ratio) in a test tube (30 mm). The solution
was allowed for decantation. Solid particles were settled well at the bottom of the
test tube (30mm). The clear liquid layer from the solution was transferred to a vial
with the aid of a dropper. The vial containing the organic solvent was placed
aside for the meantime. A pre-coated TLC plastic sheet (5x10 cm) with
fluorescent indicator was prepared. Six light pencil marks were made with 1-cm
apart and 1-cm from the bottom edge of the TLC sheet. Solvent front was made
by drawing a straight line lightly at 1-cm below the opposite end of the bottom
portion. The pre-coated TLC plastic sheet (5x10 cm) with fluorescent indicator
was applied with six different solvent samples to their respective location where
each sample had an assigned dot mark. In applying the samples to the TLC
plastic sheet with fluorescent indicator, one end of the capillary tube was dipped
in the sample solution and was allowed to enter by capillary action. Capillary
action was achieved by using an index finger to block the top-end of the capillary
tube, lightly and quickly to the spot where acetaminophen was assigned. Using
new capillary tubes, the same process was repeated for the spots where aspirin,
caffeine, sample, ibuprofen and mefenamic acid were assigned respectively.
Ethylacetate-methanol-acetic acid (25:1:1) was transferred from the vial to the
beaker. The TLC plate was carefully placed inside the beaker containing the
organic solvent. It was made sure that the height of the solvent did not reach the
spots of the TLC plate. The beaker was covered with an evaporating dish. The
solvent inside the beaker was allowed to move up until it reached the solvent
front mark on the TLC. The TLC plate was removed and air dried. The developed
TLC plate was placed inside the Ultraviolet chromatography box. The short wave
UV lamp was turned on and the spots were quickly traced by a pencil. The center
of each blot on the chromatogram was marked. The Rf value of each spot was
calculated and noted.
VI. Conclusion
The study was able to view thin layer chromatography as a versatile technique
for the examination and chemical separation of complex mixtures. Also, it truly
suits and allows one aiming to separate, identify, quantify and analyze the active
component found in commercial analgesics. Having Biogesic as a sample
used, it was found out that it has an equal Rf value with acetaminophen
(paracetamol) of about 0.48. Therefore, it can be assumed that the active
component found in Biogesic is acetaminophen (paracetamol).

VII. References
Skoog, D.A. & West, D.M. (2000). Analytical Chemistry: An Introduction. Quebec,
Canada: CE Publishing Company
[1]National Center for Biotechnology Information, U.S. National Library of
Medicine (December 1, 2010). PubMed Health. Retrieved from February 5, 2011,
from http://www.ncbi.nlm.nih.gov/pubmedhealth/PMH0000521/
[2]Grant, K. (2008, December 2). Medication Guide: Analgesic Articles. Retrieved
from February 5, 2011, from http://ratguide.com/meds/analgesics/
[3]Scott, J. (2009, June 29). Types of Analgesic Drugs. Retrieved from February
5, 2011, from http://www.articlesbase.com/medicine-articles/types-of-analgesic-
drugs-1000902.html