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Analytical Letters
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Anti-beer Evaluation of Hydrochlorothiazide and Losartan by UV Derivative Spectrophotometry

Claudio Vetuschi a; Almagrazia Giannandrea a a Farmaco Chimico Department, University of Bari, Bari, Italy

Online Publication Date: 05 January 2003 To cite this Article: Vetuschi, Claudio and Giannandrea, Almagrazia (2003) 'Anti-beer Evaluation of Hydrochlorothiazide and Losartan by UV Derivative Spectrophotometry', Analytical Letters, 36:5, 1051 - 1064 To link to this article: DOI: 10.1081/AL-120019262 URL: http://dx.doi.org/10.1081/AL-120019262

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ANALYTICAL LETTERS Vol. 36, No. 5, pp. 10511064, 2003

PHARMACEUTICAL ANALYSIS

Anti-beer Evaluation of Hydrochlorothiazide and Losartan by UV Derivative Spectrophotometry

Claudio Vetuschi* and Almagrazia Giannandrea
Farmaco Chimico Department, University of Bari, Bari, Italy

ABSTRACT
An UV derivative method for the simultaneous determination of Hydrochlorothiazide (HCT) and Losartan (LST) in the commercial forms was developed. The fourth derivative spectrum from the alcoholic solution was used. HCT can be determined by a specic signal at 330340 nm, while LST uses the signal around 280290 nm, common to both products. Into the concentration range of HCT (10.8500)103 mg mL1 and of LST (202400)103 mg mL1, for their mixture in the ratio 1:4, two intervals were identied, one for lower values, for which the absorbance grows with concentration, and one for higher values, in which the absorbance of the solution diminishes as concentration increases. The second behaviour seems

*Correspondence: Claudio Vetuschi, Farmaco Chimico Department, University of Bari, Via E. Orabona 4, 70125 Bari, Italy; E-mail: civi@farmchim.uniba.it. 1051
DOI: 10.1081/AL-120019262 Copyright & 2003 by Marcel Dekker, Inc. 0003-2719 (Print); 1532-236X (Online) www.dekker.com

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1052

Vetuschi and Giannandrea to be due to intramolecular reactions of the drugs. For both the intervals, linear regressions were obtained by correlating the spectra signals with the drug concentrations. The best results were obtained by employing the regression equations from the downward section. Key Words: Hydrochlorothiazide; Losartan; Derivative spectrophotometry; Multiple linear regression.

INTRODUCTION Losartan (LST), potassium salt of 2-n-butyl-4-chloro-5-hydroxymethyl-1-[[20 -(1H-tatrazol-5-yl)biphenyl-4-yl]methyl]imidazole, is the prototype of a new generation of nonpeptide angiotensin II receptor antagonists.[15] Hydrochlorothiazide (HCT), 6-chloro-3,4-dihydro-2H-1,2,4-benzothiadiazine-7-sulfonamide-1,1-dioxide, is a diuretic in the benzothiadiazine class. The two drugs (Fig. 1) are employed in antihypertensive pharmaceutical formulations, alone or in association.[611] The simultaneous determination of these drugs in the commercial forms is limited to a procedure based on the use of RPHPLC.[12] Apart from this, the literature contains only analytical procedures for HCT in the human plasma and urine, in the presence of LST but without any information for its quantitation.[13] The present paper proposes an UV derivative spectrophotometric method, for the simultaneous determination of the drugs, in pharmaceutics.

Figure 1.

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Evaluation of HCT and LST

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RESULTS AND DISCUSSION The examination of the spectra for pure HCT and LST in absorbance (Fig. 2) and in successive derivative orders indicates that the best analytical selectivity can be obtained in third and fourth derivatives. Particularly, the fourth derivative spectrum, in alcoholic solution (Fig. 3), exhibits a characteristic peak-trough (Pc) at 330340 nm due to HCT alone, and a peak-trough (Ps) at 280290 nm as sum of both the products. Investigations into the Linearity Range Through some reference solutions of the two pure drugs and their mixture in 1:4 ratio, in the range of: (10500)103 mg mL1 for HCT, and (402000)103 mg mL1 for LST, the relationships on Ps as function of the concentration of HCT, LST and their mixture were obtained and reported in Fig. 4. As it can be seen, these present a parabolic shape. The shift away from linearity seems to be due to intramolecular reactions which from LST lead to the formation of two condensed dimeric products.[14] For HCT too, some degradation products are reported.[1517] By examining the whole interval of concentrations, moreover, two zones could be observed. A rst, at low concentrations, in which the absorbance of the mixtures increases, and a second at high concentrations, in which the absorbance decreases. Calibration Numerous solutions of the pure drugs and their mixtures, at concentrations within the two zones above mentioned, were prepared and the derivative spectra were recorded. All the mixtures were in the HCT/LST ratio of 1:4 0.5. In order to reduce or, at least, to keep reproducible the degradation reactions, all the solutions were prepared adding amounts exactly weighted of the drugs to the stirred nal volume of solvent, without any dilution. This produces a higher statistical variability with a greater imprecision, but it makes the method more rigorous. Fig. 5 shows the obtained overlapped spectra. It appears that the position of Ps signal depends on the solution concentration, while the  value (min max) remains constant at the value of 10 nm (Fig. 6). The peak-troughs of the two selected signals were correlated with the drug concentrations (mg mL1) through linear regressions. Good agreement with the linearity was observed for the HCT concentrations

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Vetuschi and Giannandrea Figure 2. Zero-order ethanolic spectra of: HCT (-s-) 10.1 103 mg mL1; LST (-i-) 40.2 103 mg mL1; their 1:4 mixture (-u-).

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Evaluation of HCT and LST

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Figure 3.

Fourth derivative spectra of 136.2 103 mg mL1 HCT (A) and 542.9 103 mg mL1 LST (B).

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Vetuschi and Giannandrea Figure 4. Ps as function of HCT and LST concentrations, and their 1:4 mixture. HCT: (10500)103 mg mL1; LST: (202400)103 mg mL1.

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Evaluation of HCT and LST

1057 Figure 5. Overlapping of the fourth derivative spectra of 1:4 mixture of HCT (4.3466)103 mg mL1 and LST (10.81872)103 mg mL1.

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Vetuschi and Giannandrea

Figure 6. Ps position with concentration of solutions; (~) maximum; (*) right minimum.

from 4 103 to 60 103 and for the LST from 16 103 to 240 103, in upward section; and from 130 103 to 400 103 for HCT and from 500 103 to 1600 103 for LST, in downward section. All the obtained regression equations, into the two linear sections, are listed in the Table 1. Ia and Id, which correlate Pc and HCT, and IIIa and IIId as internal correlations between the Pc and Ps signals, were obtained from the pure HCT spectra. IIa, which correlate Ps with LST, was obtained from the pure LST solutions, while for the downward section, a corresponding equation was not possible, because Ps and LST at high concentrations proved to be uncorrelated (Fig. 4). IVa and IVd, which correlate Ps, Pc, and LST through multiple linear regression, were nally established from the mixture spectra.

An analogous relationship between the Ps position and LST concentration was obtained, but this is not reported here because does not improve the nal result. EXPERIMENTAL Chemicals Pure drugs, LST and HCT utilized as standards, were kindly supplied by Merck Sharp & Dohme and Sigma-Aldrich, respectively. Ethanol (95% v/v) was spectroscopic reagent grade (C. Erba).

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Evaluation of HCT and LST

Table 1. LST (mg mL1) Regression equations Ia IIa IIIa IVa Id IIId IVd HCT 1.8968Pc 0.0011 LST 1.5082Ps 0.0238 Pc 0.1033Ps 0.0056 LST 0.1381Ps 9.4835Pc 0.0036 HCT 1.2738Pc 0.0716 Pc 0.7196Ps 0.4926 LST 0.9728Ps 2.5358Pc 0.9974 16240 103

Regression equations. r 0.9995 0.9899 0.9996 0.9998 0.9866 0.9995 0.9998 Upward

HCT (mg mL1)

460 103

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130400 103

5001600 103

Downward

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Vetuschi and Giannandrea

Commercial Forms The commercial pharmaceutical forms analyzed were: LOSAZID, by Sigma-Tau; HIZAAR, by Merck-Sharp Dohme; NEO LOTAN PLUS, by Neopharmed.

Instrumentation The UV spectra were recorded over the wavelength range 400190 nm, in 10 mm silica quartz cells, using a Perkin-Elmer Lambda 15 Spectrophotometer. The instrumental conditions were xed at: scan speed 1 nm s1, response (time constant) 1 s and spectral bandwidth 1 nm. For the fourth derivative  was 8 nm.

Sample Solutions Five tablets were weighed and reduced to a ne powder, and the average of one tablet estimated. A powder amount corresponding to one tablet, accurately weighed, was transferred into a ask containing 100 mL of stirred ethanol. The suspension was recorded on UV derivative and the drugs were measured employing the Id and IVd relationships. It was veried that the excipients present in the commercial forms had no detectable eect on the amplitude of the chosen signals. The obtained results refer to one tablet.

Analytical Procedure HCT and LST (mg mL1) were determined, by using the relationships of Table 1, as follows: HCT by means of Ia and Id. LST by means of the subsequent procedures. The HCT contribution on Ps was obtained by IIIa through the Pc value and subtracted from Ps of the mixture, so as to obtain the Ps value due to LST. The resulting Ps value was used to quantify LST, through IIa. An analogous procedure does not exist for the downward section. By regression planes IVa and IVd.

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Evaluation of HCT and LST Table 2.

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Analytical results on standard mixtures by employing Id and Ivd. Found HCT 0.101 0.118 0.155 0.213 0.254 0.311 0.373 0.412 0.476 E% 0.98 0.00 0.64 0.00 1.20 1.30 1.08 1.48 0.00 LST 0.416 0.495 0.604 0.795 1.004 1.230 1.441 1.636
a

Nominal (mg mL1) HCT 0.102 0.118 0.156 0.213 0.251 0.307 0.369 0.406 0.476
a

LST 0.408 0.502 0.601 0.802 1.001 1.218 1.436 1.615 1.953

E% 2.01 1.39 0.50 0.87 0.30 0.99 0.35 1.30

a

Unappreciable.

Table 3. Nominal (mg mL ) HCT 0.203 0.203 0.203 0.255 0.262 0.231 LST 1.015 0.792 1.008 0.996 0.996 0.996 HCT (mg mL1) 0.201 0.201 0.204 0.249 0.259 0.234
1

Validation. Found % recov. 99.16 98.67 100.74 97.92 98.93 101.13 LST (mg mL1) 1.002 0.805 0.997 1.003 0.993 1.008 % recov. 98.75 101.05 98.96 100.66 99.43 101.23

Best analytical results were achieved by the second of these procedures and especially by IVd regression of downward section. For this reason the anti-Beer title has been used. The nal results got by applying the procedure with Id and IVd, on standard laboratory mixtures are reported on Table 2.

VALIDATION Synthetic mixtures, with the component levels within the range of downward section in dierent ratios, were prepared in order to validate

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Vetuschi and Giannandrea

the method proposed. The procedure was applied on these solutions and the analytical results are reported in Table 3. By means of the values of the table, the following results p were carried out. " ts= n;  0:005; were 99.4 1.3 Accuracy: percent recoveries x for HCT and 100.0 1.1 for LST. Precision: % rsd determined for 0.203 mg mL1 of HCT was 1.1 and for 0.996 mg mL1 of LST was 0.9.

CONCLUSION This work oers a simple and fast method practicable in routine analysis of title drugs in pharmaceuticals.

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