Archive of SID: Derivative Spectrophotometric Method For Determination of Losartan in Pharmaceutical Formulations
Archive of SID: Derivative Spectrophotometric Method For Determination of Losartan in Pharmaceutical Formulations
Archive of SID: Derivative Spectrophotometric Method For Determination of Losartan in Pharmaceutical Formulations
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ABSTRACT
Losartan, a highly effective blood pressure lowering agent, has been widely used for the treatment of hy-
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pertension. A fast and reliable method for the determination of losartan was highly desirable to support
formulation screening and quality control. A first derivative UV spectroscopic method was developed for
determination of losartan in the tablet dosage form. The first derivative spectrum recorded between 220
and 320 nm, and a zero-crossing technique for first derivative measurement at 232.5 nm was selected. It
is found that the selectivity and sensitivity of method to be in desirable range. In comparison with the di-
rect UV method, the first derivative UV spectroscopy has a definite through without any interference from
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UV absorbing excipients. This method is also fast and economical in comparison to the more time-
consuming HPLC method regularly used for formulation screening and quality control and can be used
routinely by any laboratory possessing a spectrophotometer with a derivative accessory. The linear con-
centration ranges were 2-50 µg/mL, (Dl = -0.0159C - 0.0056, r =0.9994, n=6). Between days CV% ≤ 2.9,
within day CV% ≤ 2.1, analytical recovery close to 98.1 % shows the suitability of the method for determi-
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Technological and scientific progress has led to the the quantification of acyclovir, celecoxib, amiloride and
development of numerous synthetic drugs. It is therefore furosemide in the presence of degradation products and
imperative to dispose of analytical methods to deter- other ingredients [10-12]. Losartan is a synthetic orally
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mine these drugs both in the quality control manufactur- active compound which binds selectively to the AT1
ing phase of the pharmaceutical formulations and their receptor (same as angiotensin II) (Fig 1). This drug was
determination in the human body [1]. Derivative UV developed as tablet dosage form for the treatment of
spectroscopy has been widely used as a tool for quanti- hypertension.
tative analysis, characterization, and quality control in At initial formulation screening stage, formulation
agricultural, pharmaceutical, and biomedical fields [2, composition was constantly varied during a highly
3]. This technique offers various advantages over the compressed time frame. A fast and reliable method for
conventional absorbency methods such as the discrimi- the dissolution and release testing of losartan was highly
nation of the sharp spectral features over the large bands desirable. Losartan has no any maximum in its normal
and the enhancement of the resolution of overlapping spectrum and therefore we can not use a wavelength for
spectra. As a result, derivative spectroscopy usually quantitative analysis at zero order spectrums. Losartan
provides much better fingerprints than the traditional has been studied and determined by several procedures
absorbency spectra [4-8]. This outstanding feature cou- such as high-performance liquid chromatography
pled with zero crossing, least square deconvolution, or (HPLC), capillary electrophoresis, and super critical
Fourier transform data processing technique has re- fluid chromatography in biological materials and tablets
ceived increasing attention in single and multi- [13-17], spectrofluorimetric in human urine [18].
component quantitative analysis of pharmaceutical drug Derivative spectrophotometric method was recom-
substances, especially in UV absorbing matrices [9]. For mended for losartan in tablet [19], but the recommended
example, derivative UV spectroscopy has been used for linear range of the method is very narrow (4-6 µg/mL)
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IJPT | January 2004 | vol. 3 | no. 1 |
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22 | IJPT | January 2004 | vol. 3 | no. 1 Ansari et al.
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Preparation of losartan standard solutions and
calibration. A stock solution containing 200 µg/mL of
Fig 1. Structure of losartan.
losartan was prepared by dissolving 0.020 g of losartan
and the results were not compared to a standard method
such as HPLC. The aim of this study was to develop an
alternative analytical method, to the more time consum-
ing HPLC method, which can be used regularly and for
formulation screening. A first derivative UV spectros-
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in doubly distilled water, then transferring into a 100 ml
calibrated flask and diluting to the mark with water.
Losartan solution containing of 20 µg/mL were tested
for stability in solution and during the actual analysis.
The behavior of the analytes remained unchanged up to
about 3 months from their preparation. Further tests of
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copy was developed to support formulation develop-
stability (i.e., over 3 months) were found unnecessary
ment of losartan in an immediate release solid dosage
and were not made. All measurements were made at
form.
room temperature. The standard solutions were prepared
by the proper dilutions of the stock standard solution
EXPERIMENTAL with doubly distilled water to reach concentration range
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of 2-50 µg/mL.
Materials
Sample solution preparation to content uniform-
All reagents used were of analytical reagent grade. ity testing. One tablet was placed into each of ten 100-
Pharmaceutical grade losartan was obtained from Het- mL volumetric flasks. 50 mL water was added. After
ero (India). Losartan tablets and the placebo product ultrasonic vibration for 10 min until the tablets were
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were manufactured by Pharmaceutical Research and dispersed in the solution, the mixture was diluted to
Development of Daru Pakhsh laboratories (Iran). volume with water and filtered (Filter membrane of 0.45
Cozaar tablets labeled to contain 25mg losartan potas- µm, Millipore, USA). After discarding few first millili-
sium manufactured by MSD (Lot No. 210464, UK) ters of the filtrate, 2 mL of filtrate transferred to a cali-
were prepared from the Shafayab Co (Iran). Acetoni- brated 25 mL volumetric flask to obtain concentration
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trile, potassium dihydrogen phosphate from Merck of 20 µg/mL. These samples were analyzed by UV and
(Darmstadt, Germany) and used as received. Doubly HPLC methods.
distilled water was used in all stages. Possible interfering effect of tablet excipients. For
the studying possible interfering effect of excipients,
Apparatus
excipients that reported to be used in tablets were ana-
Spectrophotometric analyses were performed on a lyzed by the spectrophotometric method in concentra-
Shimadzu, 2100, UV-Vis spectrophotometer, with a tion range that can be used in tablets separately and in
1.00 cm quartz cells. The optimized operating condi- combination.
tions for recording the first derivative spectra were: scan Precision assays. Losartan standard solutions were
speed, fast; spectral slit width, 2 nm; ∆λ 10 nm; and an prepared and analyzed six times within the same day to
Fig 2. Normal spectra of: A) pure losartan solution in water (2, 4, 6, 8, 10, 15, 20, 30, 40, and 50 µg/mL); B) losartan solution (20 µg/mL) pre-
pared from losartan tablet; and C) solution prepared from placebo tablet. www.SID.ir
Derivative Spectrophotometric Method for Determination of Losartan ijpt.iums.ac.ir | 23
obtain the repeatability and six times over different days sium is soluble in mobile phase and their solution was
to obtain the reproducibility. Each assay was carried out found to be stable for 2 weeks at least. As shown in Fig
on a different sample of losartan. The percentage rela- 4, at a flow rate of 1 mL/min, the retention time of
tive standard deviation (RSD %) of the data obtained losartan was 2.8 min.
was calculated. The reversed–phase HPLC method was used to pro-
Accuracy. Known concentrations of losartan were vide a proper procedure for the rapid quality control
analyzed by spectrophotometric and HPLC methods and analysis of losartan dosage forms. For quantitative
the results were compared. The samples were analyzed analysis, the analytical data for the calibration graphs
and the mean recovery as well as the repeatability was were obtained with correlation coefficients of 0.999997.
calculated on six assays for each concentration added. The good precision of the HPLC procedure was indi-
Linearity of the method. Linearity of first deriva- cated by the relative standard derivation (< 2%).
tive spectra of losartan concentration was established by The excipients (lactose, microcrystalline cellulose,
preparing one series of losartan solution ranging from 2 HPMC, CMC, PVP, corn starch, magnesium starch,
to 50µg/mL. The first derivative spectra were recorded lactose and talc) were added to the drug for recovery
using the diluents as a blank. All solutions were meas- studies according to manufacturer’s batch formula for
ured for absorbency from 220 to 320 nm. per tablets. The data shown in Table 1 indicate good
Limit of detection (LOD) and limit of quantita- accuracy and precision of the proposed procedure. The
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tion (LOQ). LOD determined by measuring the D1 detection limit (LOD) and quantification limit (LOQ)
absorbance’s at 232.5 nm of at least 25 separate base were 0.4 µg/mL and 1.5 µg/mL, respectively.
placebo tablet samples. Average and SD of blank re- Furthermore, the proposed method does not require
sponses were calculated, LOD and LOQ were estimated the elaboration of treatment and procedures, which are
by calculating of 3 SD and 10 SD of blanks, respec-
tively.
shown in Fig 2 and Fig 3. It can be seen that in Fig 2A, Limit of detection(µg/mL) 0.4
Limit of quantitation (µg/mL) 1.5
there is no peak in zero order spectrum of losartan, and Losartan tablet (taken) (mg) 25
its breakthrough wavelength at 250 nm changes with Losartan tablet (found) (%) 99.2-110.5
different concentrations. The D1 spectra (Fig 3A) have Recovery (%) 98.1
a through at 232.5 nm with a good sensitivity and line-
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arity. Spectra with higher order of derivation had lower Method Validation
sensitivity and linearity, and because of this, only first
order derivative spectra were selected for quantitative Using regression analysis the following equation
analysis. was obtained for standard calibration curve of losartan:
However, under most circumstances, pronounced in-
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Fig 3. First derivative spectra of A) pure losartan solution in water (2, 4, 6, 8, 10, 15, 20, 30, 40, and 50 µg/mL); B) losartan solution (20 µg/mL)
prepared from losartan tablet; and C) solution prepared from placebo tablet with the same concentration of excipients as in losartan tablet.
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24 | IJPT | January 2004 | vol. 3 | no. 1 Ansari et al.
Fig 4. Chromatograms of: A) losartan standard solution (20µg/mL); and B) losartan solution prepared from Cozaar tablet (20µg/mL).
error of the slope, standard error of the intercept, corre- trol analysis of losartan in dosage forms. This method
lation coefficient(r), p of the correlation coefficient were have some advantages in comparison to similar studies,
taken in Table 1. such as wide range of linearity that makes it suitable for
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The validation parameters (linearity, selectivity, re- all in vitro studies of the tablet formulations such as
covery, precision, limit of detection and limit of quanti- dissolution studies, study of the excipients interferences,
tation) were also determined (Table 1). The derivative stability studies and comparison of the method with a
spectrophotometric method is selective because the ex- validated HPLC one as a method of choice for determi-
cipients did not interfere during the determination of
losartan. Relatively small amount of CV% (2.1 and
2.9%) confirms a precision of the method, and recovery
(greater than 98%) show good accuracy. As demon-
strated, interferences do not exist between tablet excipi-
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nation of losartan in tablet. The proposed method does
not require the elaboration of treatment and procedures,
which are usually associated with chromatographic
methods. It is very efficient and offers high sample
throughput by comparison with HPLC method. There-
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ents and losartan. Therefore first derivative spectra can fore, it undoubtfully renders in-time data turnaround
be used for quantitation of the drug. Meanwhile, the D1 during formulation development. The first-derivative
spectrum displayed a trough at 232.5 nm without any order of the spectra of the losartan was found to be suit-
interference. Stability of samples prepared in water was able for determination of it in tablets. The obtained re-
studied. Results showed that samples are stable at least sults are accurate and precise and confirmed by statisti-
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for one month, and changes during sample preparation cal parameters. There was no interference of the excipi-
and time of reading are found to be negligible. For ents in the tablet. The described first derivative spectro-
quantitative analysis of the losartan tablets, ten solutions photometric method is a simple, rapid, selective, accu-
were prepared by dissolving ten individual tablets in rate, precise and an excellent alternative to HPLC
water or mobile phase and analyzed by derivative UV method for determination of the losartan in tablet or a
and HPLC procedure. The amount of the losartan was corresponding mixture.
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2002;249:117-126.
Table 2. Determination of losartan in its tablet dosage form by deriva-
tive UV spectrophotometric, and HPLC methods 2. Wang L, Asgharnejad M. Second derivative UV spectrometric
Percent of label claimed determination of simvastatin in its tablet dosage form. J Pharm
Tablet No UV Derivative HPLC Biomed Anal 2000;21:1243-1248.
1 99.2 106.9 3. Kazemipour M, Noroozian E, Tehrani MS, Mahmoudian M. A
2 102.0 103.8 new second-derivative spectrophotometric method for the de-
3 103.9 106.5 termination of permethrin in shampoo. J Pharm Biomed Anal
4 107.4 108.7 2002;30:1379-1384.
5 106.4 104.8 4. Hassan EM. Determination of Ipratropium bromide in vials
6 101.7 106.0 using kinetic and first-derivative spectrophotometric methods. J
7 108.6 106.0 Pharm Biomed Anal 2000;21:1183-1189.
8 108.3 105.5
5. El-Gindy A. First derivative spectrophotometric and LC deter-
9 108.0 107.8
mination of benoxinate HCl and its degradation products. J
10 108.6 103.4
Pharm Biomed Anal 2000;22:215-234.
Mean 105.4 105.9
SD 3.4 1.6 6. Surekha A, Jain NK. Difference spectrophotometric estimation
RSD 3.3 1.6 of amlodipine besylate. Indian drugs 2000;37(7):351-353.
7. Raggi MA, Casamenti G, Mandrioli R, Izzo G, Kenndler E.
Quantitation of olanzapine in tablets by HPLC, CZE, derivative
CONCLUSION spectrometry and linear vlotammetry. J Pharm Biomed Anal
2000;23:973-81.
In conclusion, the proposed D1- method provides 8. Karpinska J, Mikoluc B, Piotrowska J. Application of derivative
simple and sensitive method suitable for the quality con- spectrophotometry for determination of coenzyme Q10 in phar-
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Derivative Spectrophotometric Method for Determination of Losartan ijpt.iums.ac.ir | 25
maceuticals and plasma. J Pharm Biomed Anal 1998;17:1345- 15. Tatar S, Saglik S. Comparison of UV- and second derivative-
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chromatography–UV methods. Anal Chim Act 2002;466:175- neous determination of losartan and hydrochlorothiazide in
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Address correspondence to: M. Ansari, Faculty of Phar-
macy, Kerman University of Medical Sciences, Kerman, Iran
Email: mansari1345@yahoo.com
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