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Giana Mae G.

Nuncio June 11, 2018

Dane Mykel M. Quiambao
Samuel Heinrich L. Soliven
Experiment 1: UV-Vis Spectrophotometry of Ibuprofen

I. To use ultraviolet (UV) to visible light as a stimulus to ibuprofen complexed with
cobalt and measure the extent of its absorption by the analyte.
II. To estimate the concentration of ibuprofen from commercially available samples
using the Beer-Lambert Law.


In using UV-Vis spectrophotometry as a way to determine the concentration of

ibuprofen, the dosage as to which one should in-take can easily be controlled. Ibuprofen is a
nonsteroidal anti-inflammatory drug that is sold over-the-counter for the management of mild
to moderate pain, fever and inflammation. It works by blocking cyclooxygenase, the enzyme
which makes prostaglandins. Ibuprofen is sold widespread over-the-counter in 100mg and
200mg tablets for adults, while 50mg and 100mg chewable tablets, and 100mg/5mL and
40mg/mL suspensions are sold for children. Since it is used widely for the treatment of
dysmenorrhea, osteoarthritis, rheumatoid arthritis, and juvenile idiopathic arthritis, assuring
that generic brands and brands such as Advil and Motrin provide the amount of Ibuprofen
sold as claimed (1).

Overview of the Analytic Method

A. Theoretical Background
UV-Vis spectrophotometry is widely used for the quantitative determination
of a large number of inorganic, organic, and biological species (2). All organic
compounds are capable of absorbing electromagnetic radiation, because all contain
valence electrons that can be excited to higher energy levels. In this case, ibuprofen
will be the compound subjected to the UV to visible light.
Transmittance, the ratio between transmitted radiant power and incident
radiant power, and absorbance, the common logarithm of the reciprocal of
transmittance, is not normally determined inside the laboratory, but instead inside a
transparent container or a cell. After the spectrophotometer has found the absorbance
of a certain concentration of the sample, one could make use of a linear equation
relating concentration and absorbance which is rooted upon the Beer-Lambert Law:
A = εlc (Equation 1)
such that ​A refers to absorbance, ε is the molar absorptivity which is a measure of
how well a chemical species absorbs a given wavelength of light, ​l is the path length
of the cell holder which is usually 1 cm, and ​c​ is the concentration of the sample (2).
Despite this, the Beer-Lambert Law is limited in the sense that it can only
describe the absorption behavior of dilute solutions, because of solute-solvent,
solute-solute interactions or hydrogen bonding can affect the absorptivity of the
analyte (2). One case in which this occurs is when a particle affects the charge
distribution of its neighboring particles which changes the analyte’s ability to absorb
at a given wavelength.
B. Basic Instrumentation

Instruments for measuring the absorption of ultraviolet, visible, and

near-infrared radiation (NIR) are made up of one or more sources, wavelength
selectors, sample containers, radiation transducers, signal processors and readout
Generally, a spectrophotometer has a light source which is either from a
deuterium or halogen lamp. Deuterium lamps are used for the ultraviolet region and
halogen lamps are used for visible and near-infrared regions. A monochromator is
then used to separate a spectral line of interest with a specific wavelength from others
emitted by the lamp. If a spectrophotometer has that isolated spectral line passing
through by itself, it is called a single-beam spectrophotometer. If that spectral line is
split into two before entering the sample compartment, it is called a double-beam
spectrophotometer. The sample compartment contains cell holders such that their path
lengths are 1 cm most of the time and the light passes through here such that the
incident power is enters through a detector. (2)

Reagents Physical and Chemical Safety Precautions

Ibuprofen ● White, crystalline solid ● Very hazardous in case of

(C​13​H​18​O​2​) ● MW: 206.29 g/mol ingestion, of inhalation.
(3) ● Insoluble in cold water ● Hazardous in case of skin
● Slightly hazardous in case
of eye contact permeation.

Distilled ● Colorless, odorless liquid ● Non-corrosive, non-irritant,

Water (H​2​O) ● MW: 18.02 g/mol non-sensitizer and
(pH=8.5) ● BP: 100​o​C non-permeator by skin.
(4) ● Non-hazardous in case of
● Non-hazardous in case of
● Non-irritant, non-sensitizer
and non-corrosive for lungs.
● Non-corrosive to the eyes.

Cobalt ● Pink to red solid ● Very hazardous in case of

Chloride ● MW: 237.93 g/mol eye contact
Hexahydrate ● BP: 1920​o​C ● Very hazardous in case of
(CoCl​2​*6H​2​O) skin irritation
(5) ● Very hazardous in case of
inhalation and ingestion

Cyclohexane ● Colorless, chloroform-like ● Very hazardous in case of

(C​6​H​12​) odor liquid eye contact
(6) ● MW: 84.16 g/mol ● Very hazardous in case of
● BP: 1920​o​C skin irritation
● Density: 779 kg/m​3 ● Very hazardous in case of
● Insoluble in cold water. inhalation and ingestion

Benzene ● Clear liquid ● Very hazardous in case of

(C​6​H​6​) ● MW: 78.11 g/mol eye contact
(7) ● BP: 80.1​o​C ● Very hazardous in case of
● Density: 876 kg/m​3 skin irritation
● Very slightly soluble in ● Very hazardous in case of
cold water. inhalation and ingestion

Ammonium ● Colorless, ammonia-like ● Very hazardous in case of

hydroxide odor liquid eye contact
(NH​4​OH) ● MW: 35.05 g/mol ● Very hazardous in case of
(8) skin irritation
● Very hazardous in case of
inhalation and ingestion

Materials and Equipment

1 Mortar and pestle 4 100 mL volumetric flask 1 iron stand
1 funnel 1 separatory funnel with cap 1 iron ring
7 50 mL beakers 2 Stirring rods 2 Medicine droppers
500ml beaker (waste) 1 Wash bottle 2 Spatulas
4 100 mL reagent bottles 10 50 mL reagent bottles 2 10 mL pipet
1 5 mL pipet 3 aspirators UV-Vis Spectrophotometer
Kimwipes 2 Cuvettes

Methodology​ (9)
I. Calibration Curve
A. Make 100 mL 0.2 M stock cobalt chloride solution.
B. For standard solutions, dissolve 10 mg pure ibuprofen in 100 mL water
(pH=8.5) as stock.
C. Prepare five standard solutions by mixing 10 mL pure ibuprofen stock in 10
ml varying concentrations of cobalt chloride (0.01, 0.02, 0.04, 0.08, 0.1).
D. Shake standard solutions for approximately 5 minutes at room temperature.
E. Make 100 ml stock solution of benzene and cyclohexane in 2:3 ratio. Extract
each by 4 mL organic phase mixture from 2:3 benzene-cyclohexane mixture.
Place the mixture in a separatory funnel, shaken and vent frequently. The top
layer or the organic layer is then separated out of the mixture.
F. Measure in UV-Vis at 603 nm.
II. Sample Preparation and Analysis
A. Crush 10 ibuprofen tablets using a mortar and pestle.
B. Dissolve a quantity of this equal to one tablet in water (pH=8.5) and add one
drop NH​4​OH solution.
C. Transfer in 100 mL volumetric flask and fill to mark with water.
D. Make 5 10mL aliquots and add 0.01, 0.02, 0.04, 0.08, 0.1M of cobalt chloride
by standard addition.
E. Extract each by 4 mL organic phase mixture from 2:3 benzene-cyclohexane
F. Measure in UV-Vis at 603 nm and compare to calibration curve.
III. Waste Disposal
A. Dispose used organic solvents in Non-Halogenated Waste container.
B. Dispose all other waste accordingly.

(1) Ogbru O. Ibuprofen [Internet]. [Place Unknown]:; [Date Unknown]
[updated 2017 Nov 1; cited 2018 Jun 10]. Available from:
(2) Skoog DA, Holler FJ, Crouch SR. Principles of Instrumental Analysis. 6th Ed.
Boston, MA, USA: Cengage Learning; 2007.
(3) Ibuprofen MSDS [Internet]. US:; 2005 Oct 9 [updated 2013 May 21;
cited 2018 Jun 9]. Available from:
(4) Water MSDS [Internet]. US:; 2005 Oct 10 [updated 2013 May 21;
cited 2018 Jun 9]. Available from:
(5) Benzene MSDS [Internet]. US:; 2005 Oct 10 [updated 2013 May 21;
cited 2018 Jun 11]. Available from: ​
(6) Cyclohexane MSDS ​[Internet]. US:; 2005 Oct 10 [updated 2013 May
21; cited 2018 Jun 11]. Available from:
(7) Cobalt Chloride MSDS ​[Internet]. US:; 2005 Oct 9 [updated 2013
May 21; cited 2018 Jun 11]. Available from:
(8) Ammonium hydroxide MSDS ​[Internet]. US:; 2005 Oct 9 [updated
2013 May 21; cited 2018 Jun 11]. Available
(9) Jasim HH, Abed NK. Determination of Ibuprpfen in Aqueaus Solutions and
Pharmacetical Preparations by UV-VIS Spectrophotometric. Journal of Al-Nahrain
University; 2015 Jun. 18 (2):1-9. Available from:

Signature 6/11/2018