Simultaneous UV Spectrophotometric Estimation of Acebutalol Hydrochloride and Hydrochlorothiazide in Bulk and Combined Tablet Dosage Form

There is not a single analytical methods appeared in the literature for combined estimation of Acebutalol Hydrochloride and Hydrochlorothiazide in tablets dosage form. Attempts were made to develop a simple, precise and accurate Simultaneous UV spectroscopic method of Acebutalol Hydrochloride and Hydrochlorothiazide in bulk and Sectrazide tablet dosage form by using simultaneous equation method. UV spectrophotometric method was developed and validated as per ICH guidelines using methanol as mobile phase. Acebutalol Hydrochloride and Hydrochlorothiazide individually follows the Beer-Lamberts law over concentration range 3-18μg/ml and 1-6μg/ml, regression of coefficient was found to be r2=0.9999 and r2=0.9999 respectively. The percentage recovery was found in the range of 98% to 102% at three different levels. The proposed method was successfully applied for the determination of Acebutalol Hydrochloride and Hydrochlorothiazide in tablets dosage form as per ICH guidelines the result of the analysis were validated statistically and were found to be satisfactory.


Objective
The objective of the present study was to develop new analytical UV spectrophotometry method and its validation parameters for the proposed method according to ICH guidelines for the estimation of Acebutolol hydrochloride and Hydrochlorothiazide in tablets dosage form. Attempts were made to develop a simple, precise and accurate Simultaneous UV spectroscopic method.

Chemical and reagents
Acebutolol hydrochloride and Hydrochlorothiazide [bulk drug] used were of analytical reagent grade purchased from Marksons Pharmaceutical Industry, Pvt. Ltd. Verana, Goa, India, methanol (AR grade) were purchased from Research lab fine chem. Industries Mumbai and double distilled water was used throughout the analysis.

Instrumentation
A shimadzu 1800UV/VIS double beam spectrophotometer with 1cm matched quartz cells was used for all spectral measurements [5].

Preparation of standard stock solution
10mg of Acebutalol and 10mg of Hydrochlorothiazide were weighed accurately and transferred to a separate 10ml volumetric flask, dissolved in sufficient quantity of methanol then sonicated for 15min and diluted to 10 ml with the same solvent so as to get the concentration of 1000μg/ml [6].

Determination of absorption maxima
Appropriate dilution of two drugs were prepared separately using standard stock solutions containing Acebutalol Hydrochloride and Hydrochlorothiazide were scanned in the range of 400nm to 200nm to determine the wavelength of maximum absorption for both the drugs. Acebutalol Hydrochloride and Hydrochlorothiazide showed absorbance maxima at 234nm and 224nm respectively. The overlain spectra showed max of both drugs (Figure 3) [7][8][9][10][11][12]

Analysis of marketed formulation by proposed method
Ten tablets of brand name Sectrazide were used. A quantity of tablet powder equivalent to Acebutalol Hydrochloride (10mg) and Hydrochlorothiazide (10mg) was transferred to 10ml volumetric flask and dissolved in methanol. The aliquot portion of filtrate was further diluted to get Acebutalol Hydrochloride (160ug/ml) and Hydrochlorothiazide (10ug/ml) respectively (Table 1).

Method validation
The method is developed and validated according to analytical procedure as per the ICH guidelines for validation of analytical procedures. All the parameters such as linearity, precision, LOD, LOQ and accuracy for the analytes were found to be within the limit and satisfactory. The recovery studies showed that the result were within the limit indicating no interference (Table 2 & 3) (Figure 4) [13,14].    From the individual spectra of Acebutalol Hydrochloride and Hydrochlorthiazide in methanol ( Figure 5 & 6) at concentration of 10ug/ml of Acebutalol Hydrochloride and 10μg/ml Hydrochlorthiazide, two wavelengths 234nm and 224nm were selected for simultaneous estimation of drugs respectively.

Results and Discussion
The relation between concentration and absorbance for individual drug was studied. Acebutalol Hydrochloride and Hydrochlorothiazide individually follows the Beer-Lamberts law over concentration range 3-18μg/ml and 1-6μg/ml respectively. The absorptivity values for both the drugs were determined at the selected wavelengths for Acebutalol Hydrochloride and Hydrochlorthiazide respectively. Validation result is shown in the Table 4 [15,16] (Figure 7).