A

Practical Book of Physical Pharmaceutics - |I| Chemlcal Kinetlcs &Stabllty Studles

Experiment No. 09
Afm: Determinationof reaction rate constant first order.
Bequirements:
(a) Chemicals: Methyl acetate, 0.5 N hydrochloric acid, 0.1 N sodium hydroxide,
phenolphthalein indicator and distilled water.
(b) Glassware and apparatus: Beakers, pipettes, burette and its stand, reagent botles,
ound bottom flask, reflux condenser, constant temperature bath, thermometer etc.
Jheory:
The reaction rate is the speed at which a chemical reaction is taking place. Usually
reactions progress at different speeds. Rate law is an equation that relates the rate of
reaction to the concentration of reactants raise to several of power. In working the rate laws,
it is important to realize that the rate of a reaction depenids on concentration, but the rate
constant does not. The constant, k in the rate law is called the rate constant.
H*
CH,COOCH3 + H,0 ’ CH3COOH + CHOH
Methyl acetate Acetic acid Methanol
Initially, 0
At time 't a-x X X

dx
Rate of reaction = (9.1)
dt
= k[CH,COOCH3l. [H,0] (9.2)
At time ' from the start of reaction, the concentration of methyl acetate decreases and
the decreased concentration be 'x moles/litre then at any time 't change in concentration is
(a - x) moles/litre. The specific reaction rate constant 'k is calculated by using following
equation:

k =
2.303 a
t log10 a -x ".. (9.3)
The concentration of methyl acetate varies with progress of reaction (time) and hence it
is first order reaction. The objective of this experiment is to determine the rate
constant of a
reactjon by calculation and from graph.
PrÑcedure:
1. Clean and dry all the glassware.
2. Take 100 mL 0.5N HCÊ in a reagent bottle. Place the bottle in
constant temperature
water bath and note the temperature.
(Note: Temperature of bath should be maintained constant as any variation affects
rate of reaction).
3. Add 5 mL methyl acetate to above mentioned reagent bottle and
note the time at
addition as t = 0 min.
(3.12)
APractical Book of Physlcal Pharmaceutlcs -|| Chemlcal Klnetlcs &Stability Studies

4. Shake the reaction mixture and immediately withdraw a 5 ml sample from mixture
and transfer to a conical flask containing crushed ice or cold water.
(Note: Approximately 10 g crushed ice or 10 mL cold water is required to freeze the
equilibrium).
5. Titrate this mixture against 0.1N sodium hydroxide using phenolphthalein as an
indicator. Let the titre reading be Vo mL. The end point is detected when pink colour
persist for at least one minute).
6. Frequently swirl (or stir) reaction mixture and by withdrawing each time 5ml mixture
from the reagent bottle at times t = 10, 20, 30, 40, 50 min titrate as described in
step 5, above. Let the titre reading be V; m.
6. Reflux remaining reaction mixture on a boiling water bath for about an hour. After
refluxation cool the reaction mixture to water bath temperature.
7. Withdraw 5 mL of refluxed reaction mixture and titrate against 0.1 N sodium
hydroxide solution using phenolphthalein as an indicator. Let the titre reading be
Vao ml
Qbservations:
1. Water bath temperature =...°C.
2. Titre reading at t = 0, (Vo) = ... mL 0.1N NaOH.
3. Infinity reading (V.) ....mL0.1 N NaOH.
N NaOH.
4.nitial concentration of ester = a = V- V, =..... mL 0.1
Observation Table:
X= (V- VT) (a.- x) = log (a - x) k= 2.303
Time (t). Ve
(V- V) (mL)
log10 a- X
(min) (mL) (mL)

10
20
30
40
50 Mean k=... min

NaOH required at time t.

V = Volume (mL) of 0.1N
constant.
k = Reaction rate
concentration of methyl acetate.
X = Change in
Concentration of methyl acetate remaining at time (t).
a -x =
Calcutations: substitution : Calculate values of k. by substituting titre
1. Determination of k by in equation (9.3) of reaction rate constant for first
readings at different time interval
order reaction.
(3.13)
A Practlcal Book of Physical Pharmacoutlcs - || Chemlcal Kinetlcs & Stablllty Studies

2.303
k = log1o a-X
t

2. Determination of kfrom graph: Find values of 'k' from slope of the graph as given
below. In logarithmic form, the rate constant equation for first order is written as
-k
log(a - x) = t+ log a ... (9.4)
2.303
Plot the graph of log (a - x) vs. time, as
shown in Fig. 9.1 for first order reaction. (a
lòg
k):
Draw the best fit line, find slope and
calculate value of 'k' by using equation Slope = 2.303
k
Slope 2.303
k = -2.303 xslope min
Time (min.).
Fig. 9.1: Plot of log (a - x) Against Time
Precautions:
1. Maintain temperature of reaction mixture constant during reaction.
2. Shake reaction mixture after every withdrawal.
3. Use dry pipette to withdraw reaction mixture samples.
4. Withdraw samples at right time intervaland use ice or cold water to arrest reaction.
5. Withdraw samples accurately.
6.adge the end point accurately.
Result:
The reaction rate constant (k) was found to be min (by substitution) and
min from graph. Since the value of 'k' remains practically constant, the
hydrolysis of the
methyl acetate in the presence of acid is a first order reaction.
REVIEW QUESTIONS
 APractlcal Book of Physical Pharmaceutics -I|
Chemlcal Kinetics &Stablity Studles

Experiment No. 10
Aid Determtnation of reaction rate constant second order (when [a] = (b]).
Requirements:
(a) Chemicals: 0.04 Methyl acetate, 0.04 Msodium hydroxide, 0.01 Nsodium hydroxide,
0.01 N acetic acid, phenolphthalein indicator and distilled water.
(b) Glassware and apparatus: Beaker, pipette, burette, stand, reagent bottles, round
bottom flask, retlux condenser, constant temperature bath, thermometer etc.
Theory:
When the rate of reaction depends on concentration of two reactants it is called as
second order reaction. In this type of reaction rate of reaction is directly proportional to
product of concentration of the two reactants.
For example, alkaline hydrolysis of methyl acetate and hydrolysis of chlorobutanol. Let us
discuss the alkaline hydrolysis of methyl acetate
CH,CÓOCzHs + Nat OH CH,COONa + CzHsOH
Ethyl acetate Sodium hydroxide Sodium acetate Ethanol
Initially, b
At timet a -x a -x

(:: a= b)
- dx
Rate of reaction = dt
= k(CH;COOC,Hs).(NaOH)
= k(a - x) (a - x)
= k(a - x) (10.1)
sodium
At time t from the stat of reaction, the concentration of ethyl acetate and
is same
hydroxide decreases from ato (a - x). Since the concentration of both the reactants
then at any time
at the beginning (a = b) and the decreased concentration be 'x' moles/litre
from start of reaction change in concentration is (a - x) moles/litre. Integrating equation
(10.1) we get,
X
k = ... (10.2)
at (a - x)
progress of reaction
The concentrations of ethyl acetate and sodium hydroxide vary with
(time) andhence it is a second order reaction.
Procedure:
1. Clean and dry allthe glassware.
stoppered reagent bottle and 50 mL of
2. Place 50 mL of 0.04 M ethyl acetate in one
same type. Keep both the bottles in
0.04M sodium hydroxide in another bottle oftemperature.
Constant temperature water bath and note the
(3.15)
 Chemical Kinetlcs & Stabllity Studies
A Practical Book of Physical Pharmaceutics - Il
3. Transfer sodium hydroxide from second bottle to first bottle containing10ethyl acetate.
mL reaction
Quickly shake it well and note time t = 0. Immediately after, withdraw acetic acid
mixture and transfer it into conical flask, already containing 25 mL 0.01N
and 10 g ice or 10 mL cold water.
excess acetic acid
4. Titrate this mixture against 0.01N sodium hydroxide to neutralize
using phenolphthalein as an indicator. (Titre reading = Vo).
transfer into different
5. Withdraw 10 mL reaction mixture from bottle each time and
titrate
conical flasks at t =3, 6, 9, 12, 15, 20, 25 and 30 min from start of reactiòn and
separately as described in step 3 above. Let the titre readings are V
6. Determine t reading after 1 hour from the start of reaction.
7. Withdraw 10 mL of reaction mixture and transfer to conical flask containing 25 mL
0.01 Nacetic acid and 10 g ice or 10 mL cold water.
8. Titrate this mixture against 0.01N sodium hydroxide using phenolphthalein as an
indicator. Let the titre reading be Voor
(Note: This reaction is very fast even at room temperature. Therefore, V reading may
be taken after completion of one hour from the start of the reaction without
refluxing)
Qbservations:
1. Water bath temperature =...... °C.
2. Titre reading at t = 0, (V) =....mL 0.01N NaOH.
3. Infinity reading (V) =..... mL 0.01N NaOH.
4. Ahitialconcentration of ester (a = b) = (Vo- V) =... mL:0.01N NaOH.
Qbservation Table:
Time (t) X = (V- V) (a - x) =. 1
(min) (mL) (mL) (Vo-V) (mL) (a-x)

3
6
9
12
15
20
25
30

Mean k =.. L/mole.min

V, = Volume (mL) of 0.01N NaOH required at time t.
k = Specific reaction rate constant
X = Change in concentration of methyl acetate.
(a-x) = Concentration of remaining methyl acetate at time V.
(3.16)
 A
Practical Book of Physical Pharmaceutics -||
Chemical Kinetlcs &Stabillty Studies
Calculetions:
1, Determination of k by
readings at different timesubstitution
: Calculate values of k by
in equation of reaction rate substituting titre
reaction (a =b). constant for second order

2.
k
Determination of k from graph:
(10.3)
Rearranging equation (10.3), we get
k = 1
X

a(a - x)t ... (10.4)

Slope = k ... (10.5)
X
Plot the graph of
oT ala - x)vVs.
VS. time,
time, as shown in Fig. 10.1 for second order reaction.

x)ala

Slope = k

Time (min.) -
X
Figure 10.1: Plot of a(ala x)
- against Time
Draw the best fit line, find slope and calculate value of 'k' by using equation (10.5).
Precautions:
1. Maintain temperature of reaction mixture constant during reaction.
2. Shake reaction mixture after every withdrawal.
3. .Use dry pipette to withdraw reaction mixture samples.
4. Withdraw samples at right time interval and use ice or cold water to arrest reaction.
5. Withdraw samples accurately.
6. Judge the end point accurately.
Result:
.... /mole.min by substitution and
The reaction rate constant (k) was found to be hydrolysis of
L/mole.min by graph. Since the value of 'k' remains practically constant, alkaline
order reaction.
ethyl acetate (when a = b) is asecond
REVIEW QUESTIONS
A Practical Book of Physical Pharmaceutics -|| Micrometrics

Experíment No. 02
Aim:Determination of particle size, particle size distribution using microscopicmethod.
Requlrements:
(a) Chemicals: Any powder sample or diluted suspension.
(b) Glassware and apparatus: Microscope, eye-piece micrometer scale, standard stage
micrometer scale, glass slide, cover slips etc.
Theory:
The need for particle size control in the manufacture of pharmaceuticals is becoming
increasingly apparent for the solubility profiling. Precise particle size control helps in the
development of novel drug delivery platforms. Thus, the need for highly reproducible particle
size assessment techniques has grown significantly in the past decade. Microscopy method is
suitable for counting and characterizing particles of 1um and greater. With the increased
be detected and
resolving power of the microscope particles smaller than 1 um can
characterized. Microscopy method is useful for characterizing particles that are not spherical.
A microscope with sufficient magnification allows adequate characterization of the
smallest particles to be classified in the sample under examination. All the elements of the
recommendations of
optical system should be aligned and focused in accordance with the
Before use, it must be
the equipment manufacturer. Critical axial alignment is essential.
protected from vibration.
ensured that the microscope is stable and is placed on a surface
sufficiently high to allow adequate
The magnification and numerical aperture must be magnification should be determined
of the images of the particles. The actual
resolution
calibrated stage micrometer to calibrate an ocular micrometer. If the magnification is
using a
of the particle is at least 10 ocular divisions, errors can be minimized.
such that the image
Each objective should be calibrated separately.
micrometer and the ocular scale should be
To calibrate the ocular scale, the stage
order to enable a precise determination of the distance between ocular stage
aligned in necessary to characterize materials of
different magnifications may be
divisions. The use of greater
particle size in a sample. Count all the particles having a maximum dimension
varying particles define the size by diameter and for
size limit. For spherical
than the prescribed Standard.
particles, by the definitions of the different types of diameter stated in the
irregular
particles the homogeneity of the powder must be checked using
In case of irregularly shaped
descriptions of shape are acicular, columnar,
appropriate magnification. Commonly used
equant, flake, lath, plate etc. high, particularly in view of FDA trends
measurements is
The interest in particle size descriptions of particle size distributions in
thorough
toward recommending more drug product claim is based in a tightly controlled
submissions in which the emphasis of a
particle size.
(1.17)
 APractjeál Book of Physlcal Pharmaceutics - I| Micrometrics

Procedure:
1. Clean the stage, the light reflectingmirror, the eye-piece of the microscope, standard
stage micrometer scale as wellas slide, properly.
2. Adjust light for maximum intensity and mount standard stage micrometer scale on
the stage. Fit eye-piece scale in eyepiece of microscope for its calibration.
3. Calibrate eyepiece scale using standard stage micrometer scale as descritbed under
calculations and find relation of one division of eyepiece scale with respect to
micrometer unit.
4. Remove standard stage micrometer scale from stage of microscope.
5. Sprinkle small amount of the given powder on aclean slide. (Note: If emulsion or
suspension is used, just mount adrop of sample on the slide and cover it with cover
slip.)
6. Place the slide on the stage of microscope and rotate it for better resolution.
7. Observe and count minimum of 300-500 particles for their size.
Observations:
1. One division on standard stage micrometer scale = 10 um
2. Number of divisions of standard stage micrometer scale coinciding () =...
3. Number of divisions of eyepiece scale coinciding () =
4. One division of eyepiece scale = um
Observations Table:
Mean of No. of Per cent
Size range
um
size range particles particles nd nd? nd? nd.
(d) um (n) (%)
0-5 2.5
5-10 7.5
10-15 12.5
15-20 17.5
20-25 22.5
25-30 27.5
30-35 32.5
35-40 37.5
40-45 42.5
45-50 47.5
50-55 52.5
55-60 57.5

}n =
Znd =nd' =nd' =Znd=
(1.18)
A Practical Book of Physical Pharmaceutlcs - I Micrometrics

Calculations:
1. Calibration of eyepiece scale :
Standard stage micrometer is used to calibrate eye-piece scale placed in eye-piece of
microscope. It is aglass slide on which linear scale of 1.0 mm length is engraved. The
1mm is divided into 100 equal parts and one division of standard stage micrometer
is equal to 10 mm or 0.01) Fig. 2.1. Eye-piece scale is a small disc of glass. Alinear
scale of 10 mm length is engraved on glass disc at the centre, which is divided into
total ' 00 equal parts with each part equal to 0.1 mm.
Eyepiece micrometer
10 20 30 40 50 60 70 80 90 100

Stage micrometer

Fig. 2.1: Calibration of Eyepiece Scale

One division on stage micrometer scale = 10um
If Xh division on stage micrometer = yh division on eyepiece scale (2.1)
Xx 10
Then,1 division on eyepiece scale = ... (2.2)
Y
2 Arithmetic mean diameter (din). It is calculated by the equation
End .. (2.3)
din
Surface length mean diameter (d,)
ds = E nd . . (2.4)

solume surface mean diameter (d,)

End .. (2.5)
dys = E nd
Weight-moment mean diameter (dwm)
End . .(2.6)
dwm = E nd
6Surface number mean diameter (dsn)
(E nd 2 ... (2.7)
dsn = En
The volume number mean diameter (dyn)
(E nd3 .... (2.8)
dyn = Xn
shown in Fig. 2.2.
Plot the mean particle size vs. frequency distribution curve as
.19:
A Practical Book of Physlcal Pharmaceutics- I Micrometrics
40

% f30
Frequency

20

10

0 10 20 30 40 50 60 70 80 90 100
Particle size (um)
Fig. 2.2: Particle Size Distribution Curve
Precautions:
1. Use a good microscope.
2. Observe under combination of 10x eyepiece and 45x objectives.
3. Calibrate eyepiece scale properly.
4. Dilute suspension to avoid lumps or aggregation.
5. Avoid damage to standard stage micrometer.
6.Avoid entrapment of air bubble if cover slip is used.
Re_et:
1. The arithmetic mean diameter or length number mean diameter (d) =....Am
2. The volume surface nmean diameter (dvs) ....um
3. The weight-moment mean diameter is (dwm) = ...um
4. The surface length mean diameter (ds) = .... um
5. The surface number mean diameter (dsr) = ... pm
6. The volume number mean diameter (dvn = ... um
| REVIEW QUESTIONS