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Monitoring of Bisoprolol Fumarate Stability Under Different Stress Conditions
Monitoring of Bisoprolol Fumarate Stability Under Different Stress Conditions
1* 2 2
Irena Kasagiü-Vujanoviü , Biljana Janþiü Stojanoviü , Darko Ivanoviü
*
Corresponding author: kasagic.irena@gmail.com
1
University of Banja Luka–Faculty of Medicine, Department of Drug Analysis, Banja Luka, Bosnia and Herzegovina
2
University of Belgrade–Faculty of Pharmacy, Department of Drug Analysis, Belgrade, Serbia
Abstract. Stability studies of drugs by stress study is a very than in an accelerated stability study. The exact conditions
important process which is done by treating the study drug of performing forced degradation studies are chosen based
with different stress agents, with the aim to define the critical on physical and chemical characteristics of the drug [4]. In
factors affecting the stability of the drug, to accurately define this paper, forced degradation studies conducted on
the storage conditions of the drug, as well as to identify the bisporolol fumarate (BF) is presented. Degree of degrada-
resulting degradation products. In this paper, stress studies of tion was followed by previously validated Hydrophilic In-
bisoprolol fumarate were performed, in order to examine teraction Liquid Chromatography (HILIC) with UV detec-
what are the stress agents that affect its stability.For the anal-
tion [5]. In order to confirm structure of main degradation
ysis previously optimized and validated HILIC method was
product UPLC/MS/MS method was used. Literature survey
usedIt was demonstrated that oxidative stress agent has the
shown that there are no papers dealing with monitoring of
largest effect on the degradation of bisoprolol fumarate, and
BF degradation by HILIC method. This is the first time that
then the acid and base stress agent successively. Water, as a
neutral medium, and light had no significant effect on the degradation of BF was followed by HILIC which is espe-
stability of bisoprolol fumarate. During degradation under cially useful when more polar compounds appear in degra-
acid conditions impurity A was formed and it was confirmed dation process. Some previously published papers related to
with UPLC/MS/MS method. In order to more clearly define degradation of BF are given in the papers described in the
the processes of degradation, kinetic studies of degradation of references 6–9. No data on this subject.
bisoprolol fumarate have been carried out, in order to deter-
mine the order of the reaction rate of degradation and degra-
dation half-time, which provided clearer definition of the Materials and Methods
mechanism of degradation. Chromatographic system. The analysis was done on the
chromatographic system of Agilent Technologies HP1200,
Keywords: bisoprolol fumarate, stress study, HILIC, LC- consisting of HP1200 binary pump, HP1200 UV/VIS
MS/MS, kinetic studies (DAD) detector and ChemStation Software on Windows XP
for data processing.
H
OH
O H
N CH3
HOOC
CH3
CH3
,
O
O CH3 COOH
bisoprolol fumarate
bisoprolol fumarate, impurities of bisoprolol fumarate A, K For MS/MS analysis solution of BF was prepaired in
-1
and L (Sigma-Aldrich, Germany) were used. concentration of 100 ng mL and for impurities in concen-
-1
Solutions. The stock solution of BF (c = 1 mg mL ) was
prepared in acetonitrile. Working solutions of BF with con-
-1
centration of 100 ȝg mL was prepared by diluting stock
solution with an appropriate stress agent. As stress agents
were used: 0.01 M and 0.1 M sodium-hydroxide, 0.1M and
0.01M hydrochloric acid, and hydrogen-peroxide in concen-
tration of 3%, 15% and 30%. Also, testing was carried out
under neutral conditions, wherein distilled water was used
as solvent. For degradation performance under the light
influence, during the day natural daylight in combination
with artificial white light was used or only artificial white
light during night, with 12 lamps-set. The strength of one
lamp was 18W (F74-765 daylight – 1,200 lumens,
Tungsram, Hungary). Immediately, following the addition
of stress agents, analysis were carried out and chromato-
grams were developed out of samples. Then, degradation of
all BF samples, was followed after 1h, 24h, 48h and 72h,
without stopping the degradation reaction. All assays were
performed at room temperature (~25°C) analysis in the acid
medium (0.01M HCl), basic medium (0.01M NaOH) and
water were conducted at elevated temperature (50°C).
-1
The stock solutions of impurities (c = 0.1 mg mL ) was
prepared in acetonitrile, and their working solutions of 10
-1
ȝg mL concentration, were prepared by diluting with the
mobile phase.
-1
tration of 10 ng mL . All solutions were prepaired in sol-
vent consisted of 20:80 V/V methanol:water.
Table 1. The degree of degradation of BF after the stress study in a defined time interval
0.1M
6.89 8.21 11.0 14.98 24.54
NaOH
0.01M
3.01 3.39 3.42 3.86 4.20
HCl
3%
11.17 27.23 33.0 49.30 61.59
H2 O2
15%
49.19 50.44 68.93 75.01 85.11
H2 O2
30%
72.87 100.0 – – –
H2 O2
o o o
Figure 3. Chromatogram of BF after treatment with 0.01M HCl on 25 C, 50 C and 70 C
-1 -1
constant was 0.1328 mM h , while reaction half-time was
59.17h. BF oxidative degradation in 15% H2O2 is the first-
order reaction (Table 6, Fig.12). The rate constant value was
-1
0.0197h , while reaction half-time was 35.18h. From the
data obtained from degradation kinetics studies under the
action of hydrogen-peroxide, it can be seen that of all tested
stress conditions, this degradation process is the most ex-
pressed.
c (concentration)
time [h] ln (c) 1/c
[mM]
0.000 0.123 –2.094 8.118
0.300 0.123 –2.099 8.157
1.000 0.122 –2.106 8.217
24.000 0.119 –2.130 8.412
48.000 0.113 –2.184 8.879
72.000 0.101 –2.290 9.871
slope –0.0003 –0.0024 0.0217
intercept 0.1234 –2.0914 8.0838
r –0.9747 –0.9695 0.9633
Figure 9. The graphical determination of the reaction order of BF degradation in 0.1M NaOH
Figure 10. The graphical determination of the order of reaction degradation BF in 0.01M HCl
c (concentration)
time [h] ln (c) 1/c
[mM]
0.000 0.127 –2.063 7.871
0.300 0.098 –2.322 10.200
1.000 0.096 –2.345 10.434
24.000 0.090 –2.413 11.161
48.000 0.067 –2.706 14,.966
72.000 0.052 –2.965 19.398
slope –0.0008 –0.0100 0.1328
intercept 0.1074 –2.2263 9.1229
r 0.9099 0.9491 0.9637
Figure 11. The graphical determination of the reaction order of BF degradation in 3% H2O2
Figure 12. The graphical determination of the order of reaction degradation BF in 15% H2O2
Figure 13. The graphical determination of the reaction order of BF degradation under the influence of light
Concentration change with time, provides detailed de- scription of the reaction rate, but it is desirable to have a simple
measure of the reaction rate, and that is exactly the reaction half-time. The faster the reaction, the shorter the reaction half-
time, so it can be concluded that BF oxidative degradation is faster reaction in comparison with acid and alkaline degradation,
as well as in comparison with photoly- sis and degradation in water. This analysis confirmed that the most pronounced BF
degradation is with oxidative deg- radation, and degradation under the influence of light is very small.
Conclusion
Description of BF degradation under all tested conditions represents a significant source of data, which can be very useful
to analyze the impact of external factors on the quali- ty and effectiveness of this drug which is on the market mainly in the
form of tablets. In order to provide the patient safe, quality and effective medicine, it was concluded that the critical factors
that influence the stability must be re- duced to the lowest possible level. Based on detailed analy- sis, it has proven that the
oxidation and acid hydrolysis were one of the main degradation processes of BF, and that the temperature is very important
factor that accelerates its degradation. This paper also showed that the chemical ki- netics application for prediction of
pharmaceutical products
stability is of great importance and that we can obtain sig- 6. Kasagiü Vujanoviü I, Janþiü Stojanoviü B, Ivanoviü D.
nificant and reliable data on the stability by defining the Studije forsirane degradacije amlodipin-besilata i
degradation rate of active substance. For stability determi- bisoprolol-fumarata primjenom teþne hromatografije
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degradation can be predicted with great certainty. Tablets Formulated with Dicalcium Phosphate. Drug
Dev Ind Pharm. 2008;393-409
8. Induri M, Raju B, Prasad R. Validated and stability
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