Pharmaceutical Analysis 2 (PA2) Tutorial 7
Topic 7: Stability Testing
1. For EACH of the following scenario, identify the type of degradation that may occur.
Describe the degradation mechanism that you have identified.
➢ Look at structure of molecule & identify functional groups
➢ Take into consideration the formulation and environmental conditions
a) Procainamide oral solution stored at 40 °C
Chemical structure of procainamide
- Presence of amide in drug molecule → hence undergo hydrolysis
o Water in the formulation will act as a nucleophile which will attack
the electropositive carbon atom
o At higher temp of 40°C → will accelerate hydrolysis
b) An antiseptic mouthwash containing methyl salicylate in hydro-alcoholic vehicle.
Chemical structure of methyl salicylate
- Presence of ester in drug molecule → undergoes hydrolysis
o Water & alcohol in formulation will act as a nucleophile which will
attack the electropositive carbon atom
- Presence of phenol → also undergoes oxidation
o Oxygen will extract the electropositive atoms
c) Epinephrine injection
Chemical structure of epinephrine
- Presence of phenol → also undergoes oxidation
o Oxygen will extract the electropositive atoms
- Presence of chiral carbon → susceptible to racemization
Pharmaceutical Analysis 2 (PA2) Tutorial 7
2. Student A claimed that valproic acid and amphotericin B are susceptible to photolysis
when exposed to ultraviolet light. Explain if Student A is correct.
Chemical structure of valproic acid
Chemical structure of amphotericin B
- Only amophotericin B will undergo photolysis as it contains functional attached to
heavily conjugated system → alternate single & double bonds
3. Lidocaine and procaine are used as a local anesthetic. Explain why lidocaine has a longer
duration of anesthesia.
Chemical structure of lidocaine Chemical structure of procaine
- Procaine has a faster rate of degradation as the ester fg in procaine is more
susceptible to hydrolysis compared to amide fg in lidocaine
- Electronegativity: oxygen (O) = ~3.44, nitrogen (N) = ~3.04
- The carbon (C) in ester is more electropositive compared to the C in amide → C in
ester is bonded to 2 O atoms while C in amide is bonded to 1 O and 1 N atom →
hence ~6.66 vs ~6.48
- More electronegative atoms bonded to C atom = more electropositive the C atom
- More electropositive the C atom = more prone to hydrolysis
4. Drug A is currently available in the market in a tablet formulation. Student A was tasked
to develop a stable extemporaneous liquid formulation containing Drug A. Two different
liquid formulations were prepared at different pH values. The two formulations were
stored at 25 °C for one week. Drug A is predicted to undergo acid catalyzed hydrolysis in
solution and the reaction is predicted to follow the first order kinetic. The degradation
profiles of Drug A in the two formulations are shown below.
Pharmaceutical Analysis 2 (PA2) Tutorial 7
Degradation profile of drug A in formulation
1 (pH 7.4)
Ln Concentration of Drug A (mg/mL)
1.8
y = -0.0026x + 1.636
1.6 R² = 0.9437
1.4
1.2
1
0.8
0.6
0.4
0.2
0
0 20 40 60 80 100 120 140
Time (hour)
Degradation profile of drug A in formulation
2 (pH 3)
1.8
Ln Concentration of Drug A (mg/mL)
1.6
1.4
1.2 y = -0.013x + 1.5661
R² = 0.9964
1
0.8
0.6
0.4
0.2
0
0 20 40 60 80 100 120 140
Time (hour)
(a) Identify the more stable extemporaneous formulation. Explain your answer.
- Formulation 1 (pH 7.4) is more stable.
- It has a rate constant of 0.0026 → smaller than rate constant for F2 which is 0.013.
(b) Determine the shelf life (t 90 ) of Formulation 1 and 2 at 25 °C.
- Kf1 = 0.0026
- Kf2 = 0.013
- T90, f1 = 0.105/0.0026 = 40.4 hours
- T90, f2 = 8.08 hours
(c) The student suggests that the two formulations should be stored in the refrigerator (4 °C)
to enhance the stability of the extemporaneous formulations. Using the Arrhenius plot
below, determine the Ea for the degradation reaction AND estimate the shelf life (t 90 ) of
Formulation 1 at 4 °C.
[Note: Ideal gas constant: 1.987 caloriesK-1mole-1; absolute temperature for 4 °C and 25 °C are
277.15 K and 298.15 K respectively]
Pharmaceutical Analysis 2 (PA2) Tutorial 7
Arrhenius plot for Drug A degradation
6
5 y = -7142.9x + 24.5
R² = 0.9475
ln k /hour 4
0
0.0025 0.003 0.0035
1/T (K-1)
M = Ea / R
7142.9 = Ea / R
Ea = 14192.9 = 14200 calories/mole
T90, f1, 4°C = 0.105 / kf1, 4°C
kf1, 25°C = 0.0026, T25 = 298.15
kf1, 4°C = ??? , T4 = 277.15
let k1 be rate constant at 4°C (T1 = 277.15K), k2 be rate constant at 25°C (T2 = 298.15K)
ln (0.0026 / k1) = 1.815
0.0026 / k1 = e1.815
k1 = 0.0036 / e1.815 = 4.23379 x 104
hence, T90 = 0.105 / 4.23379 x 104 = 248 hrs