Practical class

Class 1.
Titration of Sodium Hydroxide with Hydrochloric acid
(Acid – Base Titration)
1. Theory
Sodium hydroxide react with carbon dioxide and absorbs water from the air, therefore it is
nonstandard solution; standardization of NaOH using 2nd standard solution of HCl, to
calculate concentration of NaOH, the equations reaction described by:

The end of the reaction is determined by a color indicator (usually phenolphthalein) as the
indicator changes its color from pink to colorless or vice versa when there is a minor
residual of either HCl or NaOH.
2. Apparatus
25 mL glass burette Wash bottle
Analytical balance, weighing paper Dropper bottle
25 mL conical flask. Medical gloves
50 mL beaker. Pasteur pipette
10 mL volumetric pipette Graduated pipette
50 mL volumetric flask Fume hood
3. Chemicals
Purified water
Sodium hydroxide powder
Phenolphthalein powder
Absolute ethanol
Concentrated hydrochloric acid
4. Procedure
- Prepare 50 mL of approximately accurately 0.1 M HCl solution from
concentrated HCl in 50 mL volumetric flask
- Prepare 50 mL of around 0.1 M NaOH in water in 50 mL volumetric flask
- Prepare indicator solution in dropper bottle
- Titrate NaOH by HCl
+ Fill in the burette with 0.1N HCl solution
+ In conical flask, accurately pipette 10 mL of 0.1 M NaOH and then add 1 – 2 drops
of indicator solution. Notice the color of the solution.
+ Titrate the solution with 0.1N HCl until the color of indicator just disappears.
+ Repeat the experiment thrice.
- Calculate the real concentration of NaOH.
 Class 2
Determination of absorption maxima and effect of solvents on absorption
maxima of organic compounds. Assay of paracetamol by UV method.
1. Theory
Ultraviolet–visible spectroscopy or ultraviolet–visible spectrophotometry (UV–Vis or
UV/Vis) refers to absorption spectroscopy or reflectance spectroscopy in part of the
ultraviolet and the full, adjacent visible regions of the electromagnetic spectrum.
Absorption spectroscopy deals with the spectroscopic techniques that measure the
absorption of radiation, as a function of frequency or wavelength, due to its interaction
with a sample. The sample absorbs energy, i.e., photons, from the radiating field. The
intensity of the absorption varies as a function of frequency, and this variation is the
absorption spectrum.
Absorption spectroscopy is employed as an analytical chemistry tool to determine the
presence of a particular substance in a sample and, in many cases, to quantify the
amount of the material present. Infrared and ultraviolet–visible spectroscopies are
particularly common in analytical applications. Absorption spectroscopy is also
employed in studies of molecular and atomic physics, astronomical spectroscopy and
remote sensing.
There are a wide range of experimental approaches for measuring absorption spectra.
The most common arrangement is to direct a generated beam of radiation at a sample
and detect the intensity of the radiation that passes through it. The transmitted energy
can be used to calculate the absorption. The source, sample arrangement and detection
technique vary significantly depending on the frequency range and the purpose of the
experiment.
Lambda max or absorption maxima (λmax)
Lambda max refers to the wavelength along the absorption spectrum where a substance
has its strongest photon absorption. Simply, the wavelength at which a substance
displays maximum absorption is called as lambda max (figure 1). Different compounds
may have very different absorption maxima and absorbances. Intensely absorbing
compounds must be examined in dilute solution (absorbance value less than 1), so that
significant light energy is received by the detector, and this requires the use of
completely transparent (non-absorbing) solvents. The most commonly used solvents
are water, ethanol, hexane and cyclohexane. Solvents having double or triple bonds, or
heavy atoms (e.g. S, Br & I) are generally avoided.
Figure 1: Typical example of unknown sample depicting absorption maxima at 279.50
nm
The Beer–Lambert
The Beer–Lambert law states that the absorbance of a solution is directly proportional
to the concentration of the absorbing species in the solution and the path length. Thus,
for a fixed path length, UV/Vis spectroscopy can be used to determine the concentration
of the absorber in a solution.
Choice of solvents
Every solvent is supposed to exhibit UV-vis absorbance cut-off wavelength. The
solvent cut-off is the wavelength below which the solvent itself absorbs all of the light.
So when choosing a solvent student has to be careful of its absorbance cut-off. If the
solvent is showing cut-off near the absorption maxima of the substance under
examination, another solvent is to be chosen.
Solvent UV Absorbance Cut-off (nm)
Water 180
Ethanol 205
Toluene 285
Dimethyl formamide 267
Acetone 329
Benzene 278
2. Apparatus
250 and 100 mL glass beakers
100 and 25 mL volumetric flasks
Whatmann filter paper
Filter funnel
Pestle and mortar
1 cm cuvette
Measuring cylinder
UV spectrometer
Analytical balance
Weighing paper
3. Chemicals
Paracetamol, Distilled water, Ethanol, 0.1M NaOH, 0.1M HCl, paracetamol tablets
4. Procedure
4.1. Prepare 0.1M NaOH and 0.1M HCl in 250 mL glass beakers.
4.2. Determine the absorption maxima of paracetamol
- Stock solution in different solvents (ethanol, 0.1M NaOH, 0.1M HCl,
water): Weigh accurately about 10 mg of Paracetamol and dissolve in sufficient
quantity of solvent (2/3 volume), in 100mL of volumetric flask, shake well to dissolve
completely and makeup the volume up to mark to prepare 0.1 mg/mL of stock solution.
Filter if necessary.
- Working solution: Pipette out 1 mL of solution from stock solution and add to
10 mL volumetric flask and make up the volume with fresh solvent to prepare 0.01
mg/mL of solution.
- Scan the solution in UV visible spectrophotometer to obtain the absorption
maxima.
4.3. Assay of paracetamol
- Establish calibration curve of paracetamol:
+ Working standard series: Using 25 mL volumetric flask, from the stock solution of
0.1 mg/mL paracetamol in 0.1M NaOH, dilute as following:
Concentration of Volume of stock Volume of distilled
paracetamol solution (mL) water (mL)
(mg/mL)
0.02 5 q.s to 25
0.04 10 q.s to 25
0.06 15 q.s to 25
0.08 20 q.s to 25
+ Measure the absorbance of 0.02, 0.04, 0.06, 0.08, 0.10 mg/mL at 257 nm using the 1
cm cuvette and establish calibration curve using 0.1M sodium hydroxide as blank.
- Weigh and powder 20 tablets. Weigh accurately a quantity of the powder
containing about 0.15 g of Paracetamol, add 50 ml of 0.1 M sodium hydroxide, dilute
with 100 ml of water, shake for 15 minutes and add sufficient water to produce 200.0
ml. Mix, filter and dilute 10.0 ml of the filtrate to 100.0 ml with water. To 10.0 ml of
the resulting solution add 10 ml of 0.1 M sodium hydroxide, dilute to 100.0 ml with
water and mix. Measure the absorbance of the resulting solution at the maximum at
about 257 nm.
- Calculate the content of paracetamol in each tablet.