Desloratadine analysis: As a pharmaceutical preparation and after accelerating ageing

Abstract

Desloratadine is a biologically active compound that is not described in the Polish Pharmacopoeia IX, hence, its study is particular important. The aim of this work was to establish a procedure for desloratadine analysis by way of HPTLC in combination with densitometry, so as to be able to ascertain its presence and degree of presence within pharmaceutical preparations. In our work, a mixture of ethyl acetate, n-butanol, ammonia and methanol was used as the mobile phase. Moreover, HPTLC plates precoated with silica gel 60F

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© 2015 Medical University of Lublin. All rights reserved 181
Curr. Issues Pharm. Med. Sci., Vol. 28, No. 3, Pages 181-185
Current Issues in Pharmacy and Medical Sciences
Formerly ANNALES UNIVERSITATIS MARIAE CURIE-SKLODOWSKA, SECTIO DDD, PHARMACIA
journal homepage: hp://www.curipms.umlub.pl/
INTRODUCTION
Antihistamine preparations are commonly sold items
in Poland’s pharmacies. Desloratadine is a substance with
antihistamine activity, and it is a ingredient found within
several very popular recent pharmaceutical preparations.
Polish Farmacopeia IX, however, does not describe any rec-
ommended method of desloratadine analysis [8]. Standard
approaches for desloratadine analysis as described in litera-
ture are liquid chromatography combined with mass spec-
trometry [2,4,9,12,13], spectrometry, spectrouorimetry and
high performance liquid chromatography [1,2,3,7,10,14].
Desloratadine analysis: as a pharmaceutical preparation
and aer accelerating ageing
K B*, M P, M M
1 Department of Analytical Chemistry, Faculty of Pharmacy, Medical University of Silesia in Katowice, Jagiellonska 4, 41-200
Sosnowiec, Poland
2 Institute of Plant Protection – National Research Institute, Pesticide Quality Testing Laboratory, Gliwicka 29, 44-153 Sosnicowice, Poland
* Corresponding author
e-mail: e-mail: bober@sum.edu.pl
tel.: + 48 32364 15 60, fax +48 32364 15 43
The aim of the work was to set out a procedure for des-
loratadine analysis utilizing high performance thin layer
chromatography combined with densitometry, as well to
establish the effect of accelerated ageing upon the contents
of solutions of desloratadine.
MATERIALS AND METHOD
Thin layer chromatography
A standard solution of desloratadine (Sigma-Aldrich,
USA) was prepared in ethanol (POCh, Poland) with con-
centration 1mg/1mL. In addition, particular dilutions of des-
loratadine standard solution were made up for the purpose
of obtaining a standard curve. HPTLC plates pre-coated
ARTICLE INFO ABSTR AC T
Received 08 January 2015
Accepted 15 September 2015 Desloratadine is a biologically active compound that is not described in the Polish
Pharmacopoeia IX, hence, its study is particular important. e aim of this work was
to establish a procedure for desloratadine analysis by way of HPTLC in combination
with densitometry, so as to be able to ascertain its presence and degree of presence
within pharmaceutical preparations. In our work, a mixture of ethyl acetate, n-butanol,
ammonia and methanol was used as the mobile phase. Moreover, HPTLC plates pre-
coated with silica gel 60F254 were also employed. e proposed method was tested and
subsequently validated. Spectrodensitometric analysis was then performed to determine
the optimal wavelength for the quantitative determination (λ=276 nm), and following
this, a quantitative analysis of desloratadine within certain pharmaceutical preparations
was performed. Our research also took into consideration an analysis of the products
of desloratadine decomposition that come about as a result of the accelerated aging of
its solutions. e employed procedure for accelerating the aging of such desloratadine
solutions consisted of heating these at 40°C and then irradiating the solution surfaces
with UV light. e changing color of these solutions aer 2 hours of exposure served
to indicate that degradation had occurred. Of note: as a result of irradiation with UV
light, desloratadine content was seen to decrease with time, declining to almost zero aer
30 hours. However, heating a solution of desloratadine alone did not induce a change
in its content. Solutions of desloratadine that had previously undergone irradiation and
heating were also analyzed to ascertain whether new substances were present. For this
purpose, the GC-MS process was employed. As a result of this procedure, the spectrum of
the solution aer aging showed the presence of several new peaks that displayed retention
several times larger and smaller than the normal desloratadine peak.
Keywords:
desloratadine,
densitometry,
HPTLC,
GC-MS,
accelerating ageing.
DOI: 10.1515/cipms-2015-0068 Unauthenticated
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Desloratadine analysis: as a pharmaceutical preparation and af ter accelerating ageing
182 Current Issues in Pharmacy and Medical Sciences
Desloratadine analysis: as a pharmaceutical preparation and aer accelerating ageing
Katarzyna Bober, Marlena Płonka, Marek Miszczyk
with silica gel (#1.05548, Merck, Germany) were used for
analysis. The plates were activated rst at a temperature
of 120°C for 30 min. These plates were then developed to
the height of 7.5 cm at room temperature. The glass chro-
matographic chamber was rst saturated with vapour of
mobile phase (a mixture of ethyl acetate – n-butanol – 25%
ammonia – methanol in volume ratio 30:7:6:7) for 30 min.
The desloratadine solution were then spotted using a glass
micro capillary (Camag, Switzerland). The composition of
the eluents was chosen experimentally.
Spectrodensitometric analysis
Spectrodensitometric analysis by way of the use of a
Camag TLC 3 densitometer was undertaken for the purpose
of establishing the maximum wavelength for the quantitative
analysis of desloratadine. This analysis was led in the range
of wavelengths between 200 and 400 nm. The slit dimen-
sion was 8.00×0.40 nm, while the scanning speed was 100
nm/s. Scanning was performed as absorption. The radiation
sources employed in this experiment were deuterium and
wolfram lamps.
Quantitative densitometric analysis
Densitometric scanning was performed by way of using
a Camag TLC 3 densitometer and the winCATS software.
The produced chromatographic plates were scanned using
wavelengths settled previously. Deuterium and wolfram
lamps were used as radiation sources.
Method validation
Method validation for desloratadine analysis was per-
formed according to ICH recommendations [6,11]. The lin-
earity range, limit of detection (LOD), limit of quantication
(LOQ), as well as precision and accuracy were determined
for the proposed method. The linearity range was estab-
lished on the basis of a previously generated standard curve.
All spotted solutions (3 µl spots) were in a range between
3.00 and 0.05 µg/spot. The mobile phase was a mixture of
ethyl acetate – n-butanol – 25% ammonia – methanol in a
volume ratio of 30:7:6:7. The analysis was done in triplicate.
The standard curve was prepared as a relationship between
area of densitometric peak [AU] and desloratadine content
[µg/spot].
Limits of detection (LOD) and quantication (LOQ) was
calculated on the base of data obtained from the calibration
curve.
The accuracy of the analysis was ascertained by calcu-
lating the percentage of the recovery. For this purpose, the
method of standard additions was employed. The analysis
was performed six times. The precision of the method was
determined on the basis of densitometric measurements,
and expressed as the value of the relative standard devia-
tion (RSD [%]).
Analysis of desloratadine in particular pharmaceutical
preparations
Using chromatographic conditions settled previously,
a quantitative analysis was undertaken of desloratadine
content in certain pharmaceutical preparations (Azomyr –
Polpharma, Poland and Aerius – Schering-Plough, Poland).
Tablets were rst ground in a mortar and placed into a ask.
Ethyl alcohol was added and then the ask was shaken for
30 minutes. The solution was subsequently ltered through
Whitmann lter pape, and the volume was made up to
10 mL to give a solution containing 1 mg of desloratadine
per 1 mL of solution. The ltrate was subsequently analysed.
Accelerating ageing
The procedure for accelerating ageing involved exposing
desloratadine solutions to UV (l=254 nm) light and tem-
peratures of 40°C. Desloratadine analysis during this phase
of the experiment was carried out in the chromatographic
conditions used previously for the analysis of standard
solution and for the investigated pharmaceutical preparations
(HPTLC plates (#1.05548, Merck, Germany) and mobile
phase ethyl acetate – n-butanol – 25% ammonia – methanol
at a volume ratio 30:7:6:7). Quantitative densitometric
analysis was led using wavelength l = 276 nm, by way of
using a Camag TLC 3 densitometer and software winCATS.
GC – MS analysis
Initial solutions of desloratadine and solutions previously
exposed to UV light and heating, were analysed by way of
employing GC – MS Gas chromatography (Agilent 7890A
Series, with mass spectrometer detector Agilent 5975C (EI-
electron impact) and Headspace Sampler G1888, as well as
ChemStation instrument control data acquisition and data
evaluation).
RESULTS AND DISCUSSION
The values characterizing the employed method of deslo-
ratadine analysis are presented in Table 1. Data are sourced
from the obtained calibration curve.
Table 1. Data characterizing the calibration curve and the method
of desloratadine analysis by use of high performance thin layer
chromatography.
linearity range [µg/spot] 3÷0.4026
intercept 2312.2400 (±79.5967)
slope 1773.0400 (±125.1260)
standard deviation s207.3230
determination coefcient R2 [%] 99.06
LOD [µg/spot] 0.2959
LOQ [µg/spot] 0.8966
accuracy (% of recovery) 100.21 (Aerius)
96.57 (Azomyr)
precision RSD [%] 2.70 (Aerius)
2.62 (Azomyr)
Figure 1 presents the ascertained spectrodensitogram
of desloratadine, whilst Figure 2 is the densitogram of the
desloratadine standard solution established by way of using
the wavelength previously settled upon.
Densitograms of desloratadine in the analyzed drugs
items were seen to be identical to the densitogram for a
standard solution of desloratadine. RF value for all analysis
was 0.53.
As a result of accelerating ageing, a change of colour of
solution (from colourless to yellowish) was observed after
2 hours of heating and exposure to UV light. This evidences
the degradation process affecting the investigated compound.
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Katarzy na Bober, Marlena Płonka, Mar ek Miszczyk
Vol. 28, No. 3, Pages 181-185 183
Due to exposure to UV light, the contents of the biologically
active compound (desloratadine) was established as decreas-
ing with time. After 30 hours, the aforementioned declined
reached to almost zero (0.035 mg/spot), whilst the initial
contents was 3 mg/spot. Heating the desloratadine solution
alone did not induce a change in content of this compound.
Indeed, after 30 hours of heating, the contents of deslorata-
dine was still about 2.940 mg/spot. The following gures
present the densitograms of desloratadine at the initial point
of analysis (Fig.2), after 30 hours of exposure to UV light
(Fig.3) and after 30 hours of heating (Fig.4). The subsequent
gure (Fig.5) is of the desloratadine content of investigated
solutions after exposure to UV light and heating.
Figure 1. Spectrodensitogram of desloratadine
Figure 2. Densitogram of desloratadine, l=276 nm
Figure 3. Densitogram of desloratadine aer 30h of exposure to
UV light, l=254 nm
Figure 3 reveals the existence of new densitogram peaks.
This indicates the presence of products that had come about
due to exposure to UV light. Only one clear peak is seen
in Figure 4. This that of desloratadine (the other peak seen
(RF=1) is in actuality, the face of the mobile phase). Figure
5 clearly indicates changes in desloratadine content as a
result of exposure to UV light.
Figure 4. Densitogram of desloratadine aer 30h of heating to a
temperature of 40°C
Figure 5. Changes in content of desloratadine following exposure
to UV light and heating
Figures 6 and 7 are comparisons of desloratadine
spectrum GC – MS in initial solution and in solution after
30 hours of exposure to UV light, λ=254 nm (Fig.6), as well
as after 30 hours of heating at a temperature of 40°C (Fig.7).
In the initial solution, we found some loratadine contami-
nation. This was identied as impurity E (Imp E) [5]. Note:
the presence of this chemical is not evident when analysis
is done by using TLC with densitometry. In our work, other
compounds were created as a result of heating, but analysis
revealed that loratadine content had decreased. GC – MS
analysis conrmed the results of the HPTLC – densitom-
etry analysis. Furthermore, it was evident that heating did
not have inuence on desloratadine content. However, GC
– MS analysis also conrmed that desloratadine content
signicantly decreases as a result of exposure to UV light.
The obtained spectrum indicates as well, that as a result of
exposure to UV light, many new compounds were created.
These are products of the degradation of desloratadine and
products of the degradation of the impurities present in the
initial solution.
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Desloratadine analysis: as a pharmaceutical preparation and af ter accelerating ageing
184 Current Issues in Pharmacy and Medical Sciences
Figure 6. Spectrum GC – MS for a standard solution of desloratadine and for a solution aer 30 hours of exposure to UV light
Figure 7. Spectrum GC-MS for a standard solution of desloratadine and for a solution aer 30 hours of heating
ACKNOWLEGMENT
This research was nanced by the Medical University of
Silesia as part of statutory research project KNW-1-013/N/3/0
and KNW-1-006/N/4/0.
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