CHEM-301

Laboratory

Fall 2021

Experiment 4:
Thermochromic
Transformation Equilibrium &
Thermodynamic Functions
Objectives

– Spectroscopy fundamentals
– UV-Vis Spectroscopy
– Solution preparation
– Rhodamine B molecule and its properties in solutions
– Kinetic study with UV-Vis spectrometer and temperature
probe
– Determination of change in absorbance with variation of
temperature
– Calculation of ΔHo, ΔSo, and ΔGo of the rhodamine B
lactone-zwitterion equilibrium
Spectroscopy

– Interaction of Radiation with a sample

– The study of molecular or atomic structure of a
substance by observation of its interaction with
electromagnetic radiation
– QUANTITATIVELY - For determining the amount
of material in a sample
– QUALITATIVELY – For identifying the chemical
structure of a sample
UV-Vis spectroscopy

– Absorption of a photon can increase molecular

energy in a variety of ways.
– In UV-Visible spectroscopy, the energy of the
photon corresponds with electronic energy
transitions of molecules and ions.
– UV-Visible spectroscopy is a type of electronic
absorption spectroscopy.
 Absorbance

– When matter absorbs ultraviolet radiation, the

electrons present in it undergo excitation.
– This causes them to jump from a ground state to an
excited state.
– The difference in the energies of the ground state
and the excited state of the electron is always equal
to the amount of ultraviolet radiation or visible
radiation absorbed by it.
– Absorbance is a very fast transition, on the order of
Jablonski diagram
10-15 seconds.
 UV-Vis spectrophotometers

– Substances can be liquids or

solids and measurements are
made with instruments called
SPECTROPHOTOMETERS
– Particular wavelength of
radiation passed through a
sample which can ABSORB
some of the radiation, Some of
the light will be absorbed and
UV Radiation – Wavelength range 220 - 380nm remaining part will be
VISIBLE Radiation – Wavelength range 380 - 780nm transmitted to the detector.
Transmittance & absorbance

– TRANSMITTED light intensity (It ) will less than the

INCIDENT light intensity (I0).
– The amount of light transmitted with respect to the
incident light is called TRANSMITTANCE (T)

𝐼𝑡
𝑇=
𝐼0

ABSORBANCE A = - log10 T
ABSORBANCE SPECTRUM

– By plotting Absorbance vs wavelength an ABSORBANCE

SPECTRUM is generated.
 THE LAWS OF SPECTROPHOTOMETRY
BEER – LAMBERT LAW
– There are two very important basic
laws and a third one which is a
combination of the two.
– LAMBERTS LAW – ABSORBANCE (A)
proportional to the PATHLENGTH (b)
of the absorbing medium.
– BEERS LAW - ABSORBANCE (A)
proportional to the
CONCENTRATION (c) of the sample.
BEER- LAMBERT LAW - ABSORBANCE
(A) proportional to b x c
𝑨 = 𝜺λ 𝒃 𝒄
The constant ελ is called the MOLAR
ABSORPTIVITY (extinction) COEFFICIENT
ελ and b are constant ABSORBANCE vs
CONCENTRATION should have linear
relationship (calibration curve)
 Rhodamine-B
Lactone-Zwitterion equilibrium

Rhodamine B (RB) is a xanthene dye that exhibits lactone– zwitterion equilibrium

(colorless) (highly colored)

Zwitterion
K= Conjugated double bonds
Lactone
absorb visible light
RB Lactone-Zwitterion equilibrium
RB has a unique property where it can change color reversibly when
subjected to a temperature change.
Effect of temperature on
equilibrium

Increase in T: Decrease in T:

– Ethanol loses its proton – Ethanol exhibits H-bond

– Interaction between donor/acceptor behavior
ethanol and rhodamine-B – Favors ethanol-
is inhibited rhodamine-B interaction
– Solvent can not stabilize – Ethanol stabilizes
charges carbonyl group
– Shifts equilibrium toward – Shifts equilibrium to
lactone zwitterion

Color intensity increases Color intensity increases

Calculation of
Thermodynamic Values

Gibbs-Helmholtz Equation

∆ G° = −R × T × ln K

Clasius-Clapeyron equation

∆S° ∆H° 1
ln K = − ×
R R T
Gibbs Free Energy change

The free energy change (DG) is a measure of the

spontaneity of a process and of the useful energy
available from it.

DGsys = DHsys - T DSsys

❖ DG < 0 for a spontaneous process

❖ DG > 0 for a nonspontaneous process
❖ DG = 0 for a process at equilibrium
Experiment

– Calibrate the UV-Vis spectrophotometer

– Prepare 50 ml 8x10-6M Rhodamine B solution and record
absorbance spectrum
– Determine maximum absoorption wavelength
– Place one of the solution in cuvette in ice bath and the other
to the hot water bath
– Place hot solution cuvette into the instrument and dip
temperature probe. Record absorbance change at maximum
absorption wavelength through cooling
– Repeat the same measurement with cold cuvette through
heating
Determination of maximum absorption
wavelength

Maximum UV-Vis Spectrum of Rhodamine B in Ethanol

0,35
absorption ?
wavelength = ? nm 0,3

0,25

0,2
Absorbance

0,15

0,1

0,05

0
400 450 500 550 600 650 700 750 800

-0,05
Wavelength (nm)
 Tracking changes in absorbance
at maximum absorbance
wavelength with temperature

Absorbance vs Temperature graph

0,7

0,6
Absorbance

0,5

0,4
y = -0,0638x + 1,8374
0,3 R² = 0,9908

0,2
17 18 19 20 21 22 23 24 25
Temperature (°C)
Calculation of the absorbance of
100% Z conformation by Beer’s Law

Calculate the absorbance of 100% Z solution from Beer's law

A=Ɛbc
Where:
A= maximum absorption at a certain wavelength
Ɛ= molar absorptivity (for pure Z it is 13.0 x 104 L/mol .cm)
b = the path the light travels in the sample (1 cm)
C= concentration of the sample (8x10-6M)
 Density of Ethanol at Various Temperatures

Calculation of Temperature
5 °C
Density (in g/cm3)
0.80300

fraction of zwitterion 10 °C 0.79900

and lactone
15 °C 0.79500
16 °C 0.79283
17 °C 0.79198
18 °C 0.79114
The Z fraction at each temperature can be 19 °C 0.79029
calculated using following equation 20 °C 0.78945
21 °C 0.78860
𝐴(𝑇 1) × ρ(𝑇𝑟𝑜𝑜𝑚 )
Z = 22 °C 0.78775

𝐴(100%) ×ρ(𝑇1 ) 23 °C 0.78691

24 °C 0.78606
Where ρ is the density of the alcohol used as 25 °C 0.78522
solvent (ethanol) at room temperature and 30 °C 0.77700

temperature T 35 °C 0.77200
40 °C 0.76800
45 °C 0.76300

Calculate the fraction of lactone 50 °C 0.75800

55 °C 0.75300
[L] = 1 - [Z] 60 °C 0.75100
Calculation of K and ΔG° for
each temperature

Zwitterion
K=
Lactone

Calculate the standard Gibbs Energy change (ΔGo) for each

temperature
∆ G° = −R × T × ln K

R: gas constant (8.3145 J/mol.K) and

T: Temperature (K)
ln K and 1/T relation

∆ G°= -R × T × lnK
∆ G°= ΔHo- TΔSo equation.
- R x T x ln K = ΔHo- TΔSo
If you solve it for ln K;
∆S° ∆H° 1
ln K = − ×
R R T
Assume, ΔSo and ΔHo are approximately constant
over the temperature range examined, a plot of ln K
vs 1/T should yield a straight line
 Calculation of ΔHo and ΔSo

ln K vs 1/T graph
∆S°
ln K = − 0,8
R
∆H° 1 0,6 y = 21743x - 74,024
× R² = 0,9863
R T 0,4

Calculate ΔHo and ΔSo 0,2

values from slope and 0

ln K

0,00336 0,00337 0,00338 0,00339 0,00340 0,00341 0,00342 0,00343 0,00344

intercept of the graph -0,2

-0,4
∆𝐻 𝑜
𝑆𝑙𝑜𝑝𝑒 = −
𝑅 -0,6
∆𝑆 𝑜 -0,8
𝐼𝑛𝑡𝑒𝑟𝑐𝑒𝑝𝑡 =
𝑅 -1
1/T (1/K)