ArticlePDF Available

Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage Forms by UFLC Method

Authors:
  • Pt CELL for Pharmaceutical Industries
Journal of PharmaSciTech
Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage
Forms by UFLC Method
1 2 3
Alaa El Sayed Ahmed Amin *, Mounir Zaky Saad , Mohamed Aly Amin Ahmed
Azelnidipine (AZL), (±)-3-[1-(diphenylmethyl) azetidin-3-yl] 5-
pro pa n -2 - yl 2- a mi no - 6- me t hy l -4 -( 3 -n itr oph en y l) -1, 4-
dihydropyridine-3,5-dicarboxylate as shown in Figure 1, is a new
dihydropyridine derivative with calcium antagonistic activity [1].
the determination of Azelnidipine includes HPLC [2-3], LC-MS method
[4-5], LC-ESI-MS [6-7], HPLC-MS-MS [8], in which two methods for
formulation and remaining for human plasma. Olmesartan medoxomil
(OLM), is (5-methyl-2-oxo-1,3-dioxol-4-yl) methyl 4-(2-hydroxy-
propan-2-yl )-2-propy l-1-{[2'- (2H-tetra zol-5-yl)biphenyl-4-
yl]methyl}-1H-imidazole-5-carboxylate as shown in Figure 1. A
literature survey revealed that Olmesartan is not yet official in any
pharmacopoeia. Several analytical methods have been reported for
the determination of Olmesartan medoxomil in biological fluids,
which includes LC-MS-MS [9], degradation product HPLC [10],
HPTLC [11] and HPLC with dissolution study [12].
Several clinical trials prove that Olmesartan medoxomil and
Azelnidipine gives better therapeutic effect in essential hypertension
rather than in single dosage form [13]. There was only one first
derivative spectrophotometric method reported for simultaneous
analysis [14] and HPLC simultaneous analysis [15].
Materials and Methods
Materials
All chemicals and reagents used were HPLC grade. Pure standards of
azelnidipine, Zhejiang Gaobang Pharmaceutical Co. Ltd., and
olmesartan medoxomil Qilu Tianhe Pharmaceutical Co. Ltd., were
obtained from Chinese. HPLC grade Methanol was purchased from
Romil. Water for chromatography was purchased from Merck,
Germany.
Chromatographic conditions
The analysis of drugs was carried out on a Shimadzu LC-20 XR,
prominence, equipped with an auto sampler (SIL-20AC XR,
Shimadzu, Japan) and PDA detector (SPD- M20A, Japan) was used
for the analysis. The data was recorded using LC-solution software. A
Phenomenex, Prodigy, ODS3, (250mm x 4.6mm, 5μm) column was
used for the analysis. A NSXX sonics ultrasonic bath (NS-A-12-7H,
Germany) was used for degassing of the mobile phase.
In this UFLC method separation was carried out using a mobile phase
Abstract
A simple, precise, sensitive and rapid reversed phase Ultra-Fast liquid chromatography method was developed for simultaneous determination of
azelnidipine and olmesartan medoxomil in Pharmaceutical Dosage form with greater precision and accuracy has been developed and validated. The
chromatographic separation was achieved by using Phenomenex, Prodigy, ODS3, 5 µm, 100 Å, (250 x 4.6 mm) analytical column with a mobile
phase consisting of methanol and water at the ratio of (85:15% v/v). The chromatographic condition was set at a flow rate of 1.5 ml/min, column
oven temperature 25°C and detector wavelength of 255 nm using a photodiode array detector. An injection volume of 10 µl was used for
2
azelnidipine and olmesartan medoxomil. The calibration curve of azelnidipine was linear with correlation coefficient (r ) = 0.9999; over a
concentration range of 1.0 - 60.0 µg/ml for; with a retention time of 6.80 min. While the calibration curve of olmesartan medoxomil was linear with
2
correlation coefficient (r ) = 0.9998; over a concentration range of 1.0 - 60.0 µg/ml for; with a retention time of 1.72 min. The recovery level of
azelnidipine and olmesartan medoxomil was 99.62% and 100.12%; respectively. The validated UFLC method was successfully applied to the
analysis of azelnidipine and olmesartan medoxomil in pharmaceutical dosage form.
Keywords: UFLC, azelnidipine, olmesartan medoxomil, method validation, pharmaceutical dosage forms.
ISSN: 2231 3788 (Print)
2321 4376 (Online)
Research Article
1Department of Analytical Chemistry, Benha University, Arab Republic of Egypt 13511
2Department of Chemistry, Zagazig University, Arab Republic of Egypt 44519
3Mepaco Pharmaceutical Company, Sheraton Heliopolis Cairo 11361
*Correspondence: asamin2005@hotmail.com (Tel. + 002/01205225223)
Introduction
Figure 1: Chemical structure for azelnidipine and olmesartan
medoxomil
The recommended dosing of Azelnidipine is 16 mg per day. A
literature survey revealed that Azelnidipine is not yet official in any
pharmacopoeia. Very few analytical methods have been reported for
Volume 6, Issue 2, 2016; Journal of PharmaSciTech
http://www.pharmascitech.in 69
Amin et al, Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage Forms by UFLC Method
consisting of HPLC grade methanol and water at the ratio of (85:15%
v/v). The mobile phase was filtered by using a 0.45 µm nylon
membrane filter. The column was maintained at a temperature of 25ºC
with column oven (CTO-20AC) and the flow rate was 1.5 ml/min.
Analysis was performed with injection volume of 10 μl using PDA
detection at 255 nm. The run time was set for 8.0 min. The optimized
chromatographic condition is shown in Table 1.
Preparation of stock and working standard solution
A 10 mg of azelnidipine and A 10 mg of olmesartan medoxomil
working standard were weighed and transferred into a 100 ml
volumetric flask. 85 ml of the methanol was added and shake on
vortex for 2 min; then was sonicated for 10 minutes. Working
standard solutions were prepared and further diluted in methanol to
contain a mixture of azelnidipine and olmesartan medoxomil in over
the linearity range from 1.0 - 60.0 µg/ml and 1.0 - 60.0 µg/ml
respectively.
Method validation
The present method of analysis was validated according to the
recommendations of ICH- 1996 and USP-30 for the parameters like
specificity, system suitability, accuracy, linearity and precision.
Specificity
It provides an indication of the selectivity and specificity of the
procedure. The method is to be selective, if the main peak is well
resoluted from any other peak by resolution of minimum 2. This could
be done injecting placebo and compare it with that of standard and
placebo spiked with standard and sample, then peak purity was
ascertained by use of PDA.
System suitability
System suitability was performed by injecting six replicates of
standard solution at 100% of the test condition at a 100% level to
verify the precision of the chromatographic system. The purposed
UFLC method permits the determination of azelnidipine and
olmesartan medoxomil in sample drug have different retention times.
System suitability data are given in Table 2.
Table 1: Optimized chromatographic conditions
Parameters Conditions
Stationary Phase
Mobile Phase
Flow Rate (ml/min)
Run Time (min)
Column Temperature ( )
Injection Volume (µl)
Detection Wavelength (nm)
Retention Time of Azelnidipine (min)
Retention Time of Olmesartan (min)
ºC
Prodigy, ODS3, 250 x 4.6 mm, 5 µm
Methanol and Water (85:15 v/v)
1.5
8.0
Ambient (25 )
10
255nm
6.80
1.72
ºC
Table 2: System suitability parameters for azelnidipine and olmesartan
medoxomil
S. No. Parameters Azelnidipine Olmesartan
1
2
3
Tailing factor
Retention Time
Theoretical plates
1.08
6.80
7044
0.86
1.72
839
Linearity
Is defined by the correlation coefficient, which should be found NLT
0.99, using peak area responses, Linearity for single point
standardization should extend to at least 20% beyond the
specification range and include the target Conc. This was performed
by preparing 7 different concentrations (2.5%, 5%, 25%, 50%, 100%,
125% and 150%), and then making 3 replicates of each concentration.
The linear working range was determined from the constructed
standard calibration curve.
Intraday Precision
This study was conducted by performing multiple analyses on a
suitable number of portions of a homogeneous sample. This was
performed by assaying multiple aliquots with the same
concentration. The analytical precision of the method was
determined by the relative standard deviation.
Inter-day Reproducibility (Method Ruggedness)
The degree of reproducibility determined by analysis of samples from
homogeneous lot of materials, under different but typical test
conditions The method is to be rugged, at any item if the pooled %RSD
of the total number of replicates that have been made in this item is
within the acceptance criteria, 3 replicates of a single sample of
powder material are used for each determination. First day: 3
replicates, on a second day: 3 replicates, then on third day: 3
replicates of freshly prepared test from the same sample are
analyzed, under the same conditions.
Accuracy
Accuracy was evaluated by spiking standard with sample solution.
The measurements are made at a concentration of standard mix,
which is found to be the target concentration, and at suitable intervals
around this point. The test samples was spiked with known quantities
of standard azelnidipine and olmesartan medoxomil using three
determinations over three concentrations level covering the specified
range. Relative recoveries of standard azelnidipine and olmesartan
medoxomil used in the standards were evaluated by comparing their
peak area with those obtained from the calibration curve equation.
Stability of Analytical solution
The stability of analytical solutions was established by injecting the
Volume 6, Issue 2, 2016; Journal of PharmaSciTech
http://www.pharmascitech.in 70
http://www.pharmascitech.in Volume 6, Issue 2, 2016; Journal of PharmaSciTech
solution and sample solution that were injected at periodic intervals
found to be the specified limit. The values are presented in the Table 3
and Table 4.
standard solution and sample solution at different time intervals up to
24 h (0, 12, and 24 h) by keeping the auto sampler temperature at
room temperature (25ºC). The % differences of peak area of standard
Table 4: Stability of standard and sample solution of olmesartan medoxomil
Time
Interval (h)
Standard Sample
Standard
Peak area
% Difference Sample
Peak area
% Difference
0
12
24
1277723
1277526
1277397
-
0.02
0.03
1277136
1276921
1276858
-
0.02
0.02
Table 3: Stability of standard and sample solution of azelnidipine
Time
Interval (h)
Standard Sample
Standard
Peak area
% Difference Sample
Peak area
% Difference
0
12
24
841726
841683
841662
-
0.01
0.01
841423
841375
841303
-
0.01
0.01
Results and Discussion
The proposed UFLC method required fewer reagents and materials,
and it is simple and less time consuming. This method could be used
in quality control test in pharmaceutical industries. The
chromatogram of azelnidipine and olmesartan medoxomil was shown
in Figure 2. There was clear resolution between azelnidipine and
olmesartan medoxomil with retention time of 6.80 and 1.72 minutes;
respectively.
Figure 2: A typical chromatogram for azelnidipine and olmesartan
medoxomil standard drug
Specificity
Generally, the specificity of a method is its suitability for the analysis
of a compound in the presence of potential impurities. Placebo,
standards, and sample test solutions were all injected at the same
wavelength of 255 nm to demonstrate the specificity of the optimized
method. A comparison of the retention times of azelnidipine and
olmesartan medoxomil in sample solutions and in the standard
solutions were exactly the same. Figures 2, 3 and 4 showed that there
were no interferences at the retention times for azelnidipine and
olmesartan medoxomil due to the placebo.
Figure 3: A typical chromatogram for azelnidipine and
olmesartan medoxomil sample drug
Figure 4: UFLC chromatogram of placebo
Therefore, the proposed method is suitable for the quantification of
the active ingredients in tablet formulation.
Amin et al, Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage Forms by UFLC Method
71
http://www.pharmascitech.in Volume 6, Issue 2, 2016; Journal of PharmaSciTech
Linearity
The response for the detector was determined to be linear over the
range of 1.0-60.0 µg/ml (1.0, 2.0, 10.0, 20.0, 40.0, 50.0 and 60.0) for
azelnidipine as shown in Figure 5 and data are shown in Table 5. The
response for the detector was determined to be linear over the range
Figure 5: Calibration curve of azelnidipine
Table 5: Statistical data of calibration curves of azelnidipine
S. No. % test
Concentration
Concentration
(µg/ml)
Average
Peak area
1
2
3
4
5
6
7
2.5
5
25
50
100
125
150
1
2
10
20
40
50
60
23094
44592
210639
421299
841858
1049211
1253045
Regression co-efficient = 0.9999
of 1.0-60.0 µg/ml (1.0, 2.0, 10.0, 20.0, 40.0, 50.0 and 60.0) for
olmesartan medoxomil as shown in Figure 6 and data are shown in
Table 6.
Figure 6: Calibration curve of olmesartan medoxomil.
Each reading was average of three determinations. They were
represented by the linear regression equation.
2
Y = 20894.15891x + 2872.55984, r = 0.9999
Azelnidipine
2
Y = 31481.83273x + 3849.89660, r = 0.9998
Olmesartan medoxomil
Slopes and intercepts were obtained by using regression equation
(Y = mx + c) and least square treatment of the results used to
confirm linearity of the method developed.
Table 6: Statistical data of calibration curves of olmesartan medoxomil
S. No. % test
Concentration
Concentration
(µg/ml)
Average
Peak area
1
2
3
4
5
6
7
2.5
5
25
50
100
125
150
1
2
10
20
40
50
60
37007
71212
317174
618424
1278919
1578673
1886715
Regression co-efficient = 0.9998
Each of the concentrations was injected in triplicate to get
reproducible response. Calibration curves were constructed by
plotting peak area versus concentration.
Table 7: Results of accuracy for azelnidipine and olmesartan medoxomil
Level (%) Amount of
drug
spiked (mg)
Azelnidipine Olmesartan
Found (mg) Recovery (%)
(n=3)
Amount of
drug
spiked (mg)
Found (mg) Recovery (%)
(n=3)
50
100
150
2.96
5.92
8.88
2.94
5.90
8.87
Average Recovery
SD
% RSD
99.29
99.66
99.91
99.62
0.312
0.313
2.88
5.76
8.64
2.89
5.77
8.63
Average Recovery
SD
% RSD
100.35
100.17
99.85
100.12
0.253
0.253
Accuracy
Accuracy was calculated by addition of standard drugs to
preanalyzed sample at 3 different concentration levels (50%, 100%
and 150%) and computing percentage recoveries. Standard limit of %
recovery study is 98 - 102 % as per ICH guideline. From the studies it
was concluded that % recovery study of azelnidipine and olmesartan
medoxomil complies with standard limit of ICH guideline. Results of
accuracy were proven by the Table 7 and % RSD is 0.313 and 0.253 of
Amin et al, Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage Forms by UFLC Method
72
http://www.pharmascitech.in Volume 6, Issue 2, 2016; Journal of PharmaSciTech
azelnidipine and olmesartan medoxomil respectively which is within
the acceptable limit (less than 2.0).
Inter-day Precision
Solution containing 40 μg/ml and 40 μg/ml of azelnidipine and
olmesartan medoxomil was prepared from their respective standard
stock solution. Analysis was replicated for 3 different days. The result
of inter-day precision studies was shown in Table 8.
Table 8: Inter -day precision data of azelnidipine and olmesartan medoxomil
Assay ( % labeled amount)
Azelnidipine Olmesartan
Sample
ID
(Day 1) (Day 2) (Day 3) (Day 1) (Day 2) (Day 3)
Sample-1
Sample-2
Sample-3
Sample-4
Sample-5
Sample-6
Average
SD
% RSD
99.23
99.22
98.62
100.02
98.99
99.22
99.22
0.459
0.463
99.44
99.36
100.11
100.15
99.97
99.45
99.75
0.368
0.369
98.88
99.14
99.61
99.55
98.77
99.31
99.21
0.344
0.347
99.12
99.18
99.88
98.45
98.96
99.33
99.15
0.468
0.472
99.62
99.23
100.06
99.65
98.98
99.45
99.50
0.373
0.375
98.98
99.13
99.53
99.41
98.74
99.42
99.20
0.305
0.308
Inter-day Reproducibility (Method Ruggedness)
Three replicates of a single sample of powder material are used for
each determination. First day: 3 replicates, on a second day: 3
replicates, then on third day: 3 replicates of freshly prepared test from
the same sample are analyzed, under the same conditions. The result
of inter-day reproducibility studies was shown in Table 9.
Table 9: Inter-day reproducibility data of azelnidipine and olmesartan medoxomil
Assay ( % labeled amount)
Azelnidipine Olmesartan
Sample
ID
(Day 1) (Day 2) (Day 3) (Day 1) (Day 2) (Day 3)
Sample-1
Sample-2
Sample-3
Average
SD
% RSD
99.44
99.26
99.58
99.43
0.160
0.161
99.56
98.36
99.26
99.06
0.624
0.630
100.12
99.56
99.72
99.80
0.288
0.289
98.86
99.46
98.60
98.97
0.441
0.446
98.89
98.78
98.58
98.75
0.157
0.159
99.56
99.49
98.66
99.24
0.501
0.504
Quantification limit
The limit of detection (LOD) and limit of quantification (LOQ) of the
developed method was determined by injecting progressively low
concentrations of the standard solutions using the developed
methods. The LOD is the lowest concentration of the analyte that can
be detected with signal to noise ratio (3:1) and LOQ is the lowest
concentration that can be quantified with acceptable precision and
accuracy with signal to noise ratio (10:1). The LOD of azelnidipine and
olmesartan medoxomil found to be 0.167 µg/ml and 0.170 µg/ml
respectively. The LOQ of azelnidipine and olmesartan medoxomil
found to be 0.50 µg/ml and 0.51 µg/ml respectively.
Stability of analytical solution
In this study, the mobile phases, the standard solutions, and the
sample solution were subjected to long term (24 h) stability studies.
The stability of these solutions was studied by performing the
experiment and looking for changes in separation, retention, and
asymmetry of the peaks which were then compared with the pattern
of the chromatogram of freshly prepared solutions
System suitability
The system suitability was determined by injecting six replicates of
the standard solutions and analyzing each active ingredient for its
peak area, peak tailing factor, resolution, number of theoretical plates,
and capacity factor. The values obtained demonstrated the suitability
of the system for the analysis of the above drug combinations System
suitability parameters might be fall within ±2% standard deviation
range during routine performance of the methods.
Conclusion
The validated UFLC method developed for the quantitative quality
control determination of azelnidipine and olmesartan medoxomil in
combination was evaluated for system suitability, specificity,
Amin et al, Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage Forms by UFLC Method
73
http://www.pharmascitech.in Volume 6, Issue 2, 2016; Journal of PharmaSciTech
linearity, range, accuracy (recovery), precision (repeatability and
intermediate precision). This method enables simultaneous
determination of azelnidipine and olmesartan medoxomil because of
good separation and resolution of the chromatographic peaks. As a
result, the proposed UFLC method could be adopted for the
quantitative quality control and routine analysis of tablet dosage form.
Acknowledgement
The authors are grateful to Mepaco-Medifood Pharmaceutical
Company (El Sharkia, Egypt) for her ultimate support for Research and
Development team.
References
[1] Oizumi K, Nishino H, Koike H, Sada T, Miyamoto M, Kimura T.
2+
Antihypertensive effects of CS-905, a novel dihydropyridine Ca channel
blocker. Jpn J Pharm 1989; 51:57-64.
[2] An H-M, Wang J-C. Determination of content and related substances of
azelnidipine by HPLC. West China J Pharm Sci 2006; 06.
[3] Pan, Y-F., Zhang, J-B., Ding, J., Wang, T-M. Determination of azelnidipine
tablets by HPLC. Qilu Pharm Affairs 2008; 07.
[4] Kawabata K, Urasaki Y. Simultaneous determination of azelnidipine and
two metabolites in human plasma using liquid chromatography-tandem mass
spectrometry. J Chromatogr B2006; 844:45-52.
[5] Kawabata K, Samata N, Urasaki Y, Fukazawa I, Uchida N, Uchida E,
Yasuhara Y. Enantioselective determination of azelnidipine in human plasma
using liquid chromatography-tandem mass spectrometry. J Chromatogr B
2007; 852:389-97.
[6] Zou J-J, Ji H-J, Zhou X-H, Zhu Y-B, Fan H-W, Xiao D-W, Hu Q.
Determination of azelnidipine by LC-ESI-MS and its application to a
pharmacokinetic study in healthy Chinese volunteers. Pharmazie 2008;
63:568-70.
Conflict of Interest: The authors confirm that this article content has
no conflict of interest.
[7] Ding L, Li L, Ma P. Determination of azelnidipine in human plasma by liquid
chromatography–electrospray ionization-mass spectrometry. J Pharm &
Biomed Anal 2007; 43:575-79.
[8] Jia J, Nan F, Liang M-Z, Yu Q, Qin Y-P, Xiang J. Determination and
pharmacokinetics of azelnidipine in human plasma by HPLC-MS-MS. Chin J
Hosp Pharm 2010; 24.
[9] Liu D, Hu P, Matsushima N, Li X, Li L, Jiang J. Quantitative determination of
olmesartan in human plasma and urine by liquid chromatography coupled to
tandem mass spectrometry. J Chromatogr B 2007; 856:190-97.
[10] Tomonori M, Hidetoshi K, Naoto F, Michinobu O, Takao K, Fumiyo K.
Identification of a degradation product in stressed tablets of olmesartan
medoxomil by the complementary use of HPLC hyphenated techniques. J
Pharm & Biomed Anal 2008; 47:553-59.
[11] Shah NJ, Suhagia BN, Shah RR, Patel NM. Development and validation of
asimultaneous HPTLC method for the estimation of olmesartan medoxomil
and hydrochlorothiazide in tablet dosage form. Indian J Pharm Sci 2007;
69:834-36.
[12] Sagirli O, Nall AO, Toker SE, Sensoy D. Simultaneous HPLC analysis of
olmesartan and hydrochlorothiazide in combined tablets and in vitro
dissolution studies. Chromatographia 2007; 66:213-18.
[13] Shimada K, Ogihara T, Saruta T, Kuramoto K, REZALT Study Group. Effects
of combination olmesartan medoxomil plus azelnidipine versus monotherapy
with either agent on 24-hour ambulatory blood pressure and pulse rate in
Japanese patients with essential hypertension: Additional results from the
REZALT study. Clin Therap 2010; 32:861-81.
[14] Patel N, Patel J. Simultaneous determination of azelnidipine and
olmesartan medoxomil by first derivative spectrophotometric method. Der
Pharm Lett 2012; 4:1080-84.
[15] Patel N, Patel J. Validated stability-indicating RP-HPLC method for the
simultaneous determination of azelnidipine and olmesartan in their combined
dosage form. Sci Pharm 2014; 82:541-54.
Amin et al, Simultaneous Determination of Azelnidipine and Olmesartan Medoxomil in Pharmaceutical Dosage Forms by UFLC Method
74
... Hence, we targeted this novel combination for the analytical study. Chromatographic (RP-HPLC) method [11] and rapid reversed phase Ultra-Fast liquid chromatography (UFLC) method for simultaneous determination of AZP and OLM in pharmaceutical dosage form has been reported with greater precision and accuracy [12] . Ramesh et al., ...
Article
An accurate, rapid and simple reversed-phase high performance liquid chromatography (RP-HPLC) bioanalytical method was developed for estimation of Azelnidipine (AZP) and Olmesartan medoxomil (OLM)in human plasma. The AZP, OLM and internal standard (IS) were extracted by liquid-liquid extraction technique. Chromatographic separation was accomplished using BDS Hypersil C18, 250 mm X 4.6 mm, 5µ analytical column. The mobile phase consisted of Acetonitrile: Water, pH adjusted with ortho-phosphoric acid in the ratio 60:40 with flow rate of 1ml/min. Detection and quantification were performed by UV detector at 256 nm. The retention times were found to be 10.05±0.02, 4.3±0.02 and 6.9±0.02 for AZP, OLM and IS respectively. The linearity range was found to be 0.5 to 12 µg /ml and 1 to 15 µg /ml for AZP and OLM respectively.The method was validated as per CDER guideline, was found to be suitable for analysis in biological fluid.
Article
Full-text available
High blood pressure, also called hypertension, is a common condition that is characterized by having a higher amount of pressure in blood vessels than normal. Hypertension (HT) is a very common disorder, particularly past middle age. It is not a disease in itself, but is an important risk factor for cardiovascular mortality and morbidity. For improvement activity of hypertension, Azelnidipine and Telmisartan newer combination in market, which is effective in Hypertension. This combination was developed to improve medication for Stage II Hypertension. Azelnidipine is Ca2+ channel blocker and chemically 3-[1-(Benzyldrylazetidin-3-yl] 5-isopropyl- 2- amino 6 methyl-4-(3-nitrophenyl)-1,4- dihydropyridine-3, 5dicarboxylate.Telmisartan is AT1-receptor blocker and Chemically2-{4-[[4-methyl-6-(1-methylbenzimidazol-2yl)-2–propylbenzimidazol-1yl] methyl] biphenyl)-benzoic acid. It provides information about different analytical method development like UV spectrophotometry, HPTLC, HPLC, and LC-MS methods reported for Azelnidipine and Telmisartan for individual and other drug combination. All reported methods found to be simple, accurate, economic, precise and reproducible in nature. This Review focuses on recent development in analytical method development for Azelnidipine and Telmisartan, and there were two methods reported for this combination as per our knowledge.
Article
Backgroound Products with multiple active substances mixed in a single dosage form are fixed-dose combinations. These are justified for a variety of reasons. These include a) increasing therapeutic efficacy, b) lowering adverse drug effects, c) pharmacokinetic advantages, d) lowering pill load, e) lowering individual drug doses, and f) lowering drug resistance development. Objective A recently approved fixed dose combination of azelnidipine (8 mg) and chlorthalidone (6.25 or 12.5 mg) is indicated to treat hypertension. Individual quantification methods for azelnidipine and chlorthalidone are available, but no practical and acceptable analytical approach for their combination has been documented. As a result, the goal of this literature review was to gather information on the numerous analytical instrumental approaches utilized to quantify azelnidipine and chlorthalidone in diverse matrices individually. The scientific community could use this information to design a new analytical method for analysing the recently approved combination. Methods Authors have explored various scientific databases to obtain information on analytical methods. Results The methods listed for azelnidipine and chlorthalidone are spectroscopy, high-performance liquid chromatography, hyphenated techniques, high-performance thin-layer chromatography, thin-layer chromatography, and a few other approaches. For azelnidipine and chlorthalidone, there were 26 and 46 research papers reported, respectively.
Article
A simple, accurate and precise method for the simultaneous determination of azelnidipine and telmisartan in bulk drug and pharmaceutical dosage has been developed by RPHPLC method. Separation was performed on a Hyperchrom ODS C18 HPLC Column (250*4.6mm) column and Buffer 0.05M Potassium dihydrogen orthophosphate (KH₂PO₄) Buffer (pH-4.0): Methanol (60:40) as a mobile phase, at a flow rate 1ml/min and UV detection wavelength 215 nm. The calibration of the method was performed by concentration range of 20-60μg/ml for telmisartan and 40-120 μg/ml for azelndipine. The validation of proposed method was carried out for accuracy, precision, ruggedness, specificity for both alzenidipine and telmisartan the method can be used for routine quality analysis of titled drug in tablet formulation.
Article
Full-text available
Azelnidipine (AZEL) is chemically (±)-3-(1-diphenylmethylazetidin3-yl) 5-isopropy12-amino-1, 4-dihydro-6-methyl-4-(3-nitrophenyl) 3,5-pyridinedicarboxylate. It is a dihydropyridine (DHP) type of calcium channel blocker (CCB) used for the treatment of hypertension. AZEL has two enantiomers due to an asymmetric carbon at the 4-position of the DHP ring. The pharmacological action of AZEL resides in the (R)-enantiomer. This is in marked contrast to other CCBs in which the (S)-enantiomer is responsible for the biological activity. The peculiar three-dimensional structure of the active enantiomer of AZEL may be related to its unique pharmacological features that are not shared by other DHPs such as long lasting reduction in blood pressure, decreased heart rate and antiatherosclerotic effect. AZEL also shows diuretic effect by increasing urine volume and thus reduction in retention of ions. Some analytical methods for the quantitative determination of Azelnidipine in pharmaceutical formulations like UV, LCMS/MS, RP-HPLC, HPLC-MS/MS, UFLC, LC-ESI-MS
Article
Full-text available
A simple, rapid, and highly selective RP-HPLC method was developed for the simultaneous determination of Azelnidipine (AZL) and Olmesartan (OLM) drug substances in the fixed dosage strength of 16 mg and 20 mg, respectively. Effective chromatographic separation was achieved using a Hypersil GOLD C18 column (150 mm × 4.6 mm internal diameter, 5 µm particle size) with a mobile phase composed of methanol, acetonitrile, and water in the ratio of 40:40:20 (by volume). The mobile phase was pumped using a gradient HPLC system at a flow rate of 0.5 mL/min, and quantification of the analytes was based on measuring their peak areas at 260 nm. The retention times for Azelnidipine and Olmesartan were about 8.56 and 3.04 min, respectively. The reliability and analytical performance of the proposed HPLC procedure were statistically validated with respect to system suitability, linearity, ranges, precision, accuracy, specificity, robustness, detection, and quantification limits. Calibration curves were linear in the ranges of 2-48 μg/mL for Azelnidipine and 2.5-60 μg/mL for Olmesartan with correlation coefficients >0.990. The proposed method proved to be selective and stability-indicating by the resolution of the two analytes from the forced degradation (hydrolysis, oxidation, and photolysis) products. The validated HPLC method was successfully applied to the analysis of AZL and OLM in their combined dosage form.
Article
Full-text available
A simple, rapid and reproducible HPLC method was developed and validated for the simultaneous determination of olmesartan (OLM) medoxomil and hydrochlorothiazide (HCT) in combined tablets. Chromatography was carried out on a 4.6mm I.D×200mm, 5μm cyano column with methanol–10mM phosphoric acid containing 0.1% triethylamine (pH 2.5, 50:50 v/v) at a flow rate of 1.0mLmin−1 and UV detector was set at 260nm. Valsartan (VAL) was used as internal standard (IS). A linear response was observed in the range of 0.2–6μgmL−1 (r 2=0.9998) for OLM and 0.1–4μgmL−1 (r 2=0.9999) for HCT, respectively. The method showed good recoveries (99.56% for OLM and 99.48% for HCT) and the relative standard deviation (RSD) values for intra- and inter-day precision were 0.70–1.59 and 0.80–2.00% for OLM and 1.20–1.37 and 1.63–1.93% for HCT, respectively. The developed method was applied successfully for quality control assay of OLM and HCT in combined tablets and in vitro dissolution studies.
Article
Full-text available
A simple, precise, accurate and rapid high performance thin layer chromatographic method has been developed and validated for the estimation of olmesartan medoxomil and hydrochlorothiazide simultaneously in combined dosage forms. The stationary phase used was precoated silica gel 60F 254 . The mobile phase used was a mixture of acetonitrile:chloroform:glacial acetic acid (7:2:0.5, v/v/v). The detection of spots was carried out at 254 nm. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found to be linear between 500 to 750 ng/spot for olmesartan medoxomil and 100 to 600 ng/spot for hydrochlorothiazide. The limit of detection and the limit of quantification for the olmesartan medoxomil were found to be 170 and 500 ng/spot, respectively and for hydrochlorothiazide 30 and 100 ng/spot, respectively. The proposed method can be successfully used to determine the drug content of marketed tablet formulation.
Article
This paper describes validated First Derivative Spectrophotometric method for the simultaneous estimation of Azelnidipine and Olmesartan medoxomil in synthetic mixture. Quantitative determination of the drugs was performed at 239.4 nm and at 217 nm (N = 1; δλ = 1) for Azelnidipine and Olmesartan medoxomil, respectively. Proposed method was evaluated for the different validation parameters. The specificity test showed that there was no interference from excipients commonly found in the commercial pharmaceutical formulations at the analytical wavelengths of Azelnidipine and Olmesartan medoxomil. Quantification was achieved ver the concentration range of 4-32 μg/ ml for Azelnidipine and Olmesartan medoxomil disoproxil fumerate. The mean recovery was 100.48 ± 1.011 and 100.70 ± 1.090 % for Azelnidipine and Olmesartan medoxomil, respectively. This method is simple, precise, sensitive and applicable for the simultaneous determination of Azelnidipine and Olmesartan medoxomil.
Article
Background: In a previously reported randomized, double-blind, parallel-group study of the efficacy and tolerability of olmesartan medoxomil (OLM) and azelnidipine (AZL) combination therapy compared with monotherapy with each agent in Japanese patients with essential hypertension (the REZALT study), the use of a combination of OLM, an angiotensin II receptor blocker, plus AZL, a dihydropyridine calcium channel blocker, was associated with significantly greater reductions in office sitting blood pressure (BP) and 24-hour ambulatory BP compared with monotherapy with either agent, and was well tolerated. Objective: This article reports the results from an a priori planned analysis and post hoc analyses of the diurnal BP and pulse rate (PR) profiles of OLM/AZL versus monotherapy with either agent from the REZALT study. Methods: Male and female Japanese outpatients with essential hypertension were eligible if they met the following inclusion criteria: age > or = 20 years; systolic BP (SBP) > or = 140 to <180 mm Hg and diastolic BP (DBP) > or = 90 to <110 mm Hg; and 24-hour ambulatory SBP/DBP > or = 135/> or = 80 mm Hg. Patients were randomly assigned to receive OLM/AZL 10/8 mg, OLM/AZL 20/16 mg, OLM 20 mg, or AZL 16 mg, once daily for 12 weeks. The effectiveness of the treatments was assessed using 24-hour ambulatory BP monitoring (ABPM) and PR, analyzed by time period (BP and PR, 24 hours, daytime [7 AM-<10 PM], nighttime [10 PM-<7 AM], and early morning [6 AM-<9 AM]; PR, morning [6 AM -<11 AM]) and dipping status at baseline (dippers [(Daytime BP - Nighttime BP)/Daytime BP > or = 10%] or nondippers [(Daytime BP - Nighttime BP)/Daytime BP <10%]). Results: A total of 867 patients were enrolled, and 862 randomized patients were included in the full analysis set (590 men, 272 women; mean age, 56.6 years). A total of 839 patients had assessable ABPM data (213, 211, 206, and 209 patients in the OLM/AZL 10/8 mg, OLM/AZL 20/16 mg, OLM, and AZL groups, respectively). No clinically significant between-group differences were observed in baseline demographic and clinical characteristics. Combination therapy was associated with significantly greater antihypertensive effects on 24-hour ABPM compared with either monotherapy in all of the time periods, as follows: SBP/DBP reductions with OLM/AZL 20/16 mg in the daytime, nighttime, and early morning were -22.6/-14.1, -21.2/-12.5, and -20.6/-11.9 mm Hg, respectively (all, P < 0.05 vs the other 3 treatment groups). The SBP/DBP reductions with OLM/AZL 10/8 mg (daytime, -18.2/-11.0 mm Hg; nighttime, -18.1/-10.0 mm Hg; and early morning, -15.6/-9.3 mm Hg) were also significantly greater than with OLM 20 mg (-11.8/-6.7, -12.8/-7.2, and -11.0/ -6.9 mm Hg, respectively; all, P < 0.01) and AZL 16 mg (-13.1/-7.8, -10.2/-5.5, and -9.9/-6.1 mm Hg; all, P < 0.001) in all of the time periods. The antihypertensive effects associated with OLM/AZL 10/8 mg or 20/16 mg were significantly greater than those with monotherapies regardless of dipping pattern at baseline (all, P < 0.05) in all of the time periods, with the exception of nighttime reduction with OLM/AZL 10/8 mg versus OLM in dippers. The numbers of patients who had any increase in BP were 12/213 (5.6%) with OLM/AZL 10/8 mg, 13/211 (6.2%) with OLM/AZL 20/16 mg, 35/206 (17.0%) with OLM, and 36/209 (17.2%) with AZL. The AZL-containing regimens were associated with reduced morning PR (mean [95% CI] changes from baseline to week 12: -1.5 beats/min [-2.5 to -0.4] with OLM/AZL 10/8 mg, -2.1 beats/min [-3.0 to -1.1] with OLM/AZL 20/16 mg, 0.4 beat/min [-0.5 to 1.3] with OLM, and -1.9 beats/min [-2.8 to -1.0] with AZL). Conclusion: In this study in Japanese patients with essential hypertension, the reductions in daytime, nighttime, and early-morning BP assessed using 24-hour ABPM were significantly greater with combination OLM/AZL than with either monotherapy, regardless of dipping pattern at baseline. Japan Pharmaceutical Information Center registration number: JapicCTI-060286.
Article
A simple, rapid and sensitive high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) assay for determination of azelnidipine in human plasma using perospirone as the internal standard (IS) was established. After adjustment to a basic pH with sodium hydroxide solution, plasma samples were extracted with diethyl ether and separated on a C18 column with a mobile phase of methanol-5 mM ammonium acetate solution (90:10, v/v). The lower limit of quantification (LLOQ) was 0.20 ng/ml. After administration of a single dose of azelnidipine 8mg and 16 mg, respectively; the area under the plasma concentration versus time curve from time 0 h to 96 h (AUC(0-96) were (186 +/- 47) ng ml(-1) h, (429 +/- 145) ng ml(-1) h, respectively; clearance rate (CL/F) were (45.94 +/- 11.61), (42.11 +/- 14.23) L/h, respectively; peak plasma concentration Cmax were (8.66 +/- 1.15), (19.17 +/- 4.13) ng/ml, respectively; apparent volume of distribution (Vd) were (1749 +/- 964), (2480 +/- 2212) L, respectively; time to Cmax (Tmax) were (2.8 +/- 1.2), (3.0 +/- 0.9) h, respectively; elimination half-life (t(1/2beta)) were (22.8 +/- 2.4), (23.5 +/- 4.2) h, respectively; and MRT were (25.7 +/- 1.3), (26.2 +/- 2.2) h, respectively; The essential pharmacokinetic parameters after oral multiple doses (8 mg, q.d.) were as follows: (Cmax) ss, (15.04 +/- 2.27) ng/ml; (Tmax) ss, (2.38 +/- 0.92) h; (Cmin) ss, (3.83 +/- 0.94) ng/ml; C(av), (7.05 +/- 1.54) ng/ml; DF, (1.62 +/- 0.26); AUCss, (169.19 +/- 36.87) ng ml(-1) h.
Article
A quantitative assay method by liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) for the simultaneous determination of azelnidipine and its two metabolites, M-1 (aromatized form) and M-2 (hydroxylated form), in human plasma was developed and validated. Plasma samples, each of 1.0mL, were extracted by a single step liquid-liquid extraction using a mixture of ethyl acetate and hexane (1:1, v/v), and analyzed by the LC/ESI-MS/MS method. Three analytes were separated by isocratic elution on a C(18) column, and ionized using a positive ion electrospray ionization source. The ion transitions were monitored in selected reaction monitoring (SRM) mode. The chromatographic run time was 11min per injection, with retention time of 3.6, 10.2 and 6.8min for azelnidipine, M-1 and M-2, respectively. The calibration curves for azelnidipine, M-1 and M-2 well fitted to equations by a weighted (1/X(2)) quadratic regression over the range of 0.5-40.0ng/mL (r(2)>0.9979). The intra- and inter-assay precisions (coefficient of variation: C.V.), calculated from quality control (QC) samples, were less than 8.7 and 8.4%, 3.8 and 4.7%, and 11.9 and 13.9%, respectively, for azelnidipine, M-1 and M-2. The accuracy was within +/-9% for azelnidipine, within +/-7% for M-1 and within +/-16% for M-2. The overall recoveries for azelnidipine, M-1 and M-2 were 68.8-78.6%, 54.3-62.9% and 80.4-89.7%, respectively. All analytes evaluated demonstrated acceptable short-term, long-term, auto-sampler and stock solution stabilities. Furthermore, the method developed was successfully applied to pharmacokinetic studies on azelnidipine, M-1 and M-2 after an oral dose of 16mg CALBLOCK tablets (2mgx8mg tablets) to healthy volunteers.
Article
A liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method for the determination of azelnidipine in human plasma was established. Nicardipine was used as the internal standard (IS). After adjustment to a basic pH with sodium hydroxide solution (0.1 M), plasma samples were extracted with cyclohexane-diethyl ether (1:1, v/v) and separated on a C(18) column with a mobile phase of 20 mM ammonium acetate solution-methanol-formic acid (25:75:0.5, v/v). The electrospray ionization was employed in a single quadrupole mass spectrometer for the determination. The method was linear over the concentration range of 0.05-40 ng/ml. The lower limit of quantification (LLOQ) was 0.05 ng/ml. The intra- and inter-run standard deviations were less than 9.5% and 11.0%, respectively. The method was successfully applied to study the pharmacokinetics of azelnidipine in healthy Chinese volunteers.
Article
A sensitive and simple method was developed for determination of the enantiomers of azelnidipine, (R)-(-)-azelnidipine and (S)-(+)-azelnidipine, in human plasma using chiral liquid chromatography with positive ion atmospheric pressure chemical ionization tandem mass spectrometry. Plasma samples spiked with stable isotope-labeled azelnidipine, [(2)H(6)]-azelnidipine, as an internal standard, were processed for analysis using a solid-phase extraction in a 96-well plate format. The azelnidipine enantiomers were separated on a chiral column containing alpha(1)-acid glycoprotein as a chiral selector under isocratic mobile phase conditions. Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode, monitoring the transitions from m/z 583-->167 for (R)-(-)-azelnidipine and (S)-(+)-azelnidipine, and from m/z 589-->167 for [(2)H(6)]-azelnidipine. The standard curve was linear over the studied range (0.05-20 ng/mL), with r(2)>0.997 using weighted (1/x(2)) quadratic regression, and the chromatographic run time was 5.0 min/injection. The intra- and inter-assay precision (coefficient of variation), calculated from the assay data of the quality control samples, was 1.2-8.2% and 2.4-5.8% for (R)-(-)-azelnidipine and (S)-(+)-azelnidipine, respectively. The accuracy was 101.2-117.0% for (R)-(-)-azelnidipine and 100.0-107.0% for (S)-(+)-azelnidipine. The overall recoveries for (R)-(-)-azelnidipine and (S)-(+)-azelnidipine were 71.4-79.7% and 71.7-84.2%, respectively. The lower limit of quantification for both enantiomers was 0.05 ng/mL using 1.0 mL of plasma. All the analytes showed acceptable short-term, long-term, auto-sampler and stock solution stability. Furthermore, the method described above was used to separately measure the concentrations of the azelnidipine enantiomers in plasma samples collected from healthy subjects who had received a single oral dose of 16 mg of azelnidipine.
Article
A specific, sensitive and fast method based on high performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) was developed for the determination of olmesartan in human plasma and urine. Solid-phase extraction (SPE) was used to isolate the compounds from biological matrix followed by injection of the extracts onto a C(18) column with isocratic elution. The method was validated over the concentration range of 0.2-1000 and 5-10,000 ng/mL for olmesartan in human plasma and urine, respectively. The method was applied to the pharmacokinetic study of olmesartan medoxomil in healthy Chinese male and female subjects.