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\title{Spectroscopic Estimation of Pioglitazone Hydrochloride}
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\begin{document}

             \author[1]{Dr. A. P.  Kulkarni}

             \author[2]{Shahnawaz  Mohd}

             \author[3]{Zahid  Zaheer}

             \affil[1]{  College of Pharmacy, Rauza Bagh, Aurangabad}

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\date{\small \em Received: 13 January 2012 Accepted: 10 February 2012 Published: 22 February 2012}

\maketitle


\begin{abstract}
        


A simple, sensitive, accurate, precise, reproducible and cost effective UV spectroscopic method has been developed for the estimation of Pioglitazone hydrochloride in bulk and tablet dosage form. Pioglitazone hydrochloride shows maximum absorption at 269 nm with molar absorptivity of 9.6013 × 104 l/mol.cm. Beer?s law was obeyed in the concentration range of 10-70 ?g/ml. The method was validated for linearity, precision, accuracy, sensitivity and specificity. The data obtained was treated with the statistical approach. The proposed method was found to be accurate and precise for estimation of Pioglitazone hydrochloride in bulk and tablet dosage form.

\end{abstract}


\keywords{Pioglitazone hydrochloride, UV spectrophotometeric, validation, dissolution test, quality control test}

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\let\tabcellsep& 	 	 		 \par
glucose, decreases withdrawal of glucose from the liver, and reduces quantity of glucose. \hyperref[b1]{[2]} .HCl According to literature review, a HPLC method for the estimation of Pioglitazone hydrochloride is available. \hyperref[b2]{[3]} The method is relatively complex and expensive. The UV method for estimation of Pioglitazone hydrochloride in methanol: water: hydrochloric acid (250:250:1) system \hyperref[b3]{[4]} and in 0.2 M sulphuric acid \hyperref[b4]{[5]} have been reported. However, quantitative estimation of PH in other media has not been reported. This is essential in drug release study. The objective of the study was to develop a simple, accurate, precise, cost effective and reproducible UV method for estimation of PH in 0.1N hydrochloric acid as per ICH guidelines. \hyperref[b5]{[6]} Shimadzu UV/Visible double beam spectrophotometer and a Jasco V-630 instrument with 1 cm matched quartz cells were used for the spectral measurement. Shimadzu AX200 analytical balance was used for the weighing purpose. The reference standard of PH was obtained as a gift sample from Aarti Drugs, Thane (India) with 99.8\% assay value. PH tablets (Piomed, 15 mg) were obtained from the market and utilized for the study. All other chemicals were of analytical grade. 
\section[{a) Selection of The Media}]{a) Selection of The Media}\par
The criterion for selection of the medium was the solubility and the stability, i.e. PH should be soluble Standard solution of PH was prepared by dissolving 100 mg of drug in 100 ml of 0.1N hydrochloric acid (Solution A, 1000 µg/ml). Further 10 ml of the solution A was diluted to 100 ml with 0.1N hydrochloric acid (Solution B, 100 µg/ml). Solution B was used as the standard stock solution. 
\section[{c) Preparation of Calibration Curve}]{c) Preparation of Calibration Curve}\par
Aliquots of 1 ml to 7 ml of the standard solution B were transferred into a series of calibrated 10 ml standard volumetric flasks and the final volume was made up using 0.1N hydrochloric acid. The solutions were scanned in the range of 200-400 nm against blank (0.1N hydrochloric acid). The absorption maximum was found to be at 269 nm. (Figure  {\ref 2})The absorbance of the solutions were measured at 269 nm against the blank (Table \hyperref[tab_0]{1}) and the calibration curve was constructed. (Figure  {\ref 3})  The proposed method was applied to marketed PH tablets (Piomed, 15 mg). Twenty tablets of PH were weighed and powdered in a glass mortar. Powder equivalent to 100 mg of the drug was weighed accurately and transferred to a 100 ml standard volumetric flask. It was dissolved in about 50 ml of 0.1N hydrochloric acid and the volume was made up with 0.1N hydrochloric acid so that the concentration was 1000 µg/ml (Solution P). Ten ml of the solution P was transferred to a 100 ml standard volumetric flask and the volume was adjusted with 0.1N hydrochloric acid (Solution Q). The solution was filtered through Whatmann filter paper no. 41. The filtrate was diluted suitably with 0.1N hydrochloric acid to obtain a sample solution (20µg/ml). The absorbance of the sample solution was measured at 269 nm and the amount of PH was determined from the calibration curve. The method was studied for accuracy and precision. a) Linearity Pioglitazone hydrochloride exhibited maximum absorption at 269 nm and obeyed Beer's Law in the range of 10-70 µg/ml. \hyperref[b7]{[8,}\hyperref[b9]{10]} Linear regression of absorbance Vs concentration yielded equation y= 0.022x + 0.017 with a correlation coefficient of 0.999. 
\section[{b) Accuracy}]{b) Accuracy}\par
To determine the suitability and reproducibility of the proposed method, recovery studies were carried precision, the \% drug content and the relative standard deviation (RSD) values 99.59722 ± 0.4722, 100.7488 ± 0.4522, 100.4226 ± 0.5617 and 0.4940 respectively. When the analyst was changed the RSD values were 0.48225 and 0.4662. According to ICH guidelines, an acceptance criterion for the precision is RSD ? 2\%.\par
out by adding known amount of standard PH (80\%, 100\%, and 120\%) to the tablet solution P and analyzing the mixtures by the proposed method. Three samples were prepared for each recovery level. The percentage recovery of PH was found to be 99.3233 ± 0.7026 (Table \hyperref[tab_2]{3}) indicating that there is no interference by the excipients in the method. According to ICH guidelines, an acceptance criterion for the \% recovery is 98-102\%.  
\section[{c) Precision}]{c) Precision}\par
Precision of the method was demonstrated by intra-day and inter-day variation studies. For intra-day precision, six sample solutions of Pioglitazone hydrochloride of same concentration (20µg/ml) were analyzed three times in a day. The result is indicated by \% RSD in Table \hyperref[tab_3]{4}.\par
During the intermediate precision (inter-day precision), six sample solutions of the same concentration (20µg/ml) were analyzed on three consecutive days and by two different analysts in same laboratory. The results are indicated by \% RSD in Table \hyperref[tab_5]{5 and 6}.\par
For intra-day precision, the \% drug content and the relative standard deviation (RSD) were found to be 99.958 ± 0.7874, 99.928 ± 1.104, 99.297 ± 1.114 and 1.0087 respectively; whereas for inter-day  When the analysis was carried on two different instruments, the RSD values were 0.5297 and 0.5213. The LOD and LOQ of PH were determined by using standard deviation of the response and the slope approach as defined in the ICH Guidelines \hyperref[b5]{[6]}. The LOD and LOQ were found to be 0.03µg/ml and 0.1µg/ml respectively. The proposed method showed molar absorptivity of 9.6013 × 104 l/mol.cm. (Table \hyperref[tab_1]{2)}  
\section[{March}]{March}\begin{figure}[htbp]
\noindent\textbf{1}\includegraphics[]{image-2.png}
\caption{\label{fig_0}Figure 1 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{}\includegraphics[]{image-3.png}
\caption{\label{fig_1}A}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{23}\includegraphics[]{image-4.png}
\caption{\label{fig_2}Figure 2 :Figure 3 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{1} \par 
\begin{longtable}{P{0.058465608465608464\textwidth}P{0.15740740740740738\textwidth}P{0.23835978835978833\textwidth}P{0.3957671957671957\textwidth}}
Sr.\tabcellsep Concentration\tabcellsep Absorbance\tabcellsep Standard\\
no\tabcellsep (µg/ml)\tabcellsep \tabcellsep deviation\\
1\tabcellsep 0\tabcellsep 0\tabcellsep 0\\
2\tabcellsep 10\tabcellsep 0.2380\tabcellsep ± 0.003551\\
3\tabcellsep 20\tabcellsep 0.4620\tabcellsep ± 0.003404\\
4\tabcellsep 30\tabcellsep 0.7385\tabcellsep ± 0.003593\\
5\tabcellsep 40\tabcellsep 0.9034\tabcellsep ± 0.002524\\
6\tabcellsep 50\tabcellsep 1.1134\tabcellsep ± 0.000917\\
7\tabcellsep 60\tabcellsep 1.3600\tabcellsep ± 0.001000\\
8\tabcellsep 70\tabcellsep 1.5359\tabcellsep ± 0.002571\end{longtable} \par
 
\caption{\label{tab_0}Table 1 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{2} \par 
\begin{longtable}{P{0.10602150537634408\textwidth}P{0.3454838709677419\textwidth}P{0.398494623655914\textwidth}}
Sr\tabcellsep Parameter\tabcellsep Result\\
no\tabcellsep \tabcellsep \\
1.\tabcellsep Absorption maxima\tabcellsep 269 nm\\
2.\tabcellsep Linearity range\tabcellsep 10-70 µg/ml\\
3.\tabcellsep Standard Regression\tabcellsep y = 0.022x + 0.017\\
\tabcellsep Equation\tabcellsep \\
4.\tabcellsep Correlation\tabcellsep 0.999\\
\tabcellsep coefficient (r 2 )\tabcellsep \\
5.\tabcellsep Molar Absorptivity\tabcellsep 9.6013 × 10 4 l/mol.cm\\
6.\tabcellsep A (1\%, 1 cm)\tabcellsep 244.328 dl/gm/cm\\
7.\tabcellsep Accuracy (\%\tabcellsep 99.3233 ± 0.7026\\
\tabcellsep recovery ± S.D)\tabcellsep \\
8.\tabcellsep Specificity\tabcellsep A 20 µg/ml of drug in\\
\tabcellsep \tabcellsep 0.1 N HCl at UV\\
\tabcellsep \tabcellsep detection wavelength\\
\tabcellsep \tabcellsep of 269 nm shows an\\
\tabcellsep \tabcellsep absorbance value of\\
\tabcellsep \tabcellsep 0.4620 ± 0.003404\\
9.\tabcellsep LOD (µg/ml))\tabcellsep 0.03\\
10.\tabcellsep LOQ (µg/ml)\tabcellsep 0.10\\
\multicolumn{3}{l}{d) Preparation of Sample Solution}\end{longtable} \par
 
\caption{\label{tab_1}Table 2 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{3} \par 
\begin{longtable}{P{0.45915178571428567\textwidth}P{0.17075892857142858\textwidth}P{0.13660714285714287\textwidth}P{0.08348214285714285\textwidth}}
Ingredient\tabcellsep \multicolumn{3}{l}{Pioglitazone hydrochloride}\\
Tablet amount\tabcellsep 20\tabcellsep 20\tabcellsep 20\\
(µg/ml)\tabcellsep \tabcellsep \tabcellsep \\
Level of\tabcellsep 80\tabcellsep 100\tabcellsep 120\\
addition (\%)\tabcellsep \tabcellsep \tabcellsep \\
Amount added\tabcellsep 16\tabcellsep 20\tabcellsep 24\\
(µg/ml)\tabcellsep \tabcellsep \tabcellsep \\
Amount\tabcellsep 35.748\tabcellsep 39.574\tabcellsep 44.1288\\
recovered\tabcellsep \tabcellsep \tabcellsep \\
(µg/ml)\tabcellsep \tabcellsep \tabcellsep \\
\% Recovery\tabcellsep 99.3000\tabcellsep 98.9350\tabcellsep 100.2927\\
Average \%\tabcellsep \tabcellsep 99.3233 ± 0.7026\tabcellsep \\
recovery\tabcellsep \tabcellsep \tabcellsep \end{longtable} \par
 
\caption{\label{tab_2}Table 3 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{4} \par 
\begin{longtable}{P{0.10286624203821657\textwidth}P{0.3979299363057325\textwidth}P{0.19219745222929938\textwidth}P{0.15700636942675159\textwidth}}
\tabcellsep \multicolumn{2}{l}{hydrochloride}\tabcellsep \\
Sample\tabcellsep \multicolumn{3}{l}{Analysis of Pioglitazone}\\
Number\tabcellsep \multicolumn{3}{l}{hydrochloride as percent of drug}\\
\tabcellsep \tabcellsep content\tabcellsep \\
\tabcellsep 10:00 am\tabcellsep 2:00 pm\tabcellsep 6:00 pm\\
1\tabcellsep 101.214\tabcellsep 98.979\tabcellsep 98.569\\
2\tabcellsep 99.458\tabcellsep 99.568\tabcellsep 100.598\\
3\tabcellsep 99.587\tabcellsep 99.259\tabcellsep 99.454\\
4\tabcellsep 100.254\tabcellsep 101.871\tabcellsep 100.598\\
5\tabcellsep 98.979\tabcellsep 99.298\tabcellsep 97.995\\
6\tabcellsep 100.256\tabcellsep 100.598\tabcellsep 98.568\\
Mean ±\tabcellsep 99.958 ±\tabcellsep 99.928 ±\tabcellsep 99.297\\
SD\tabcellsep 0.7874\tabcellsep 1.104\tabcellsep ± 1.114\\
Average\tabcellsep \multicolumn{3}{l}{99.7276 ± 1.00875}\\
± RSD\tabcellsep \tabcellsep 1.0087\tabcellsep \end{longtable} \par
 
\caption{\label{tab_3}Table 4 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{5} \par 
\begin{longtable}{P{0.0964179104477612\textwidth}P{0.36791044776119397\textwidth}P{0.22074626865671643\textwidth}P{0.16492537313432837\textwidth}}
\tabcellsep \multicolumn{2}{l}{hydrochloride}\tabcellsep \\
Sample\tabcellsep \multicolumn{3}{l}{Analysis of Pioglitazone hydrochloride}\\
number\tabcellsep \multicolumn{3}{l}{as percent of labeled content}\\
\tabcellsep DAY-1\tabcellsep DAY-2\tabcellsep DAY-3\\
1\tabcellsep 99.8467\tabcellsep 100.725\tabcellsep 100.053\\
2\tabcellsep 100.0230\tabcellsep 101.146\tabcellsep 99.8792\\
3\tabcellsep 99.4538\tabcellsep 99.9875\tabcellsep 99.9103\\
4\tabcellsep 98.9985\tabcellsep 100.5473\tabcellsep 101.163\\
5\tabcellsep 99.1356\tabcellsep 100.856\tabcellsep 100.5409\\
6\tabcellsep 100.1257\tabcellsep 101.231\tabcellsep 100.9892\\
Mean ±\tabcellsep 99.59722 ±\tabcellsep 100.7488 ±\tabcellsep 100.4226 ±\\
SD\tabcellsep 0.4722\tabcellsep 0.4522\tabcellsep 0.5617\\
Average\tabcellsep \tabcellsep \multicolumn{2}{l}{100.2562 ± 0.4940}\\
± RSD\tabcellsep \tabcellsep 0.4940\tabcellsep \end{longtable} \par
 
\caption{\label{tab_4}Table 5 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{6} \par 
\begin{longtable}{P{0.1516728624535316\textwidth}P{0.4929368029739777\textwidth}P{0.2053903345724907\textwidth}}
\tabcellsep \multicolumn{2}{l}{(Intra-day precision)}\\
Sample\tabcellsep \multicolumn{2}{l}{Analysis of Pioglitazone}\\
number\tabcellsep \multicolumn{2}{l}{hydrochloride as percent of}\\
\tabcellsep \multicolumn{2}{l}{labeled amount}\\
\tabcellsep Analyst-I\tabcellsep Analyst-II\\
1\tabcellsep 100.2346\tabcellsep 99.1035\\
2\tabcellsep 100.9812\tabcellsep 99.1418\\
3\tabcellsep 99.8754\tabcellsep 98.7460\\
4\tabcellsep 100.0213\tabcellsep 99.2435\\
5\tabcellsep 99.5381\tabcellsep 97.9924\\
6\tabcellsep 100.1509\tabcellsep 98.6356\\
Mean\tabcellsep 100.1335\tabcellsep 98.8149\\
Std.\tabcellsep 0.48225\tabcellsep 0.4662\\
Deviation\tabcellsep \tabcellsep \\
d) Robustness\tabcellsep \tabcellsep \end{longtable} \par
 
\caption{\label{tab_5}Table 6 :}\end{figure}
 \begin{figure}[htbp]
\noindent\textbf{7} \par 
\begin{longtable}{P{0.27247386759581876\textwidth}P{0.402787456445993\textwidth}P{0.17473867595818815\textwidth}}
\tabcellsep Instruments)\tabcellsep \\
Sample\tabcellsep \multicolumn{2}{l}{Analysis of Pioglitazone}\\
number\tabcellsep \multicolumn{2}{l}{hydrochloride as percent of}\\
\tabcellsep \multicolumn{2}{l}{labeled content}\\
\tabcellsep Shimadzu\tabcellsep Jasco\\
1\tabcellsep 99.184\tabcellsep 100.231\\
2\tabcellsep 98.793\tabcellsep 101.104\\
3\tabcellsep 99.862\tabcellsep 100.863\\
4\tabcellsep 100.021\tabcellsep 99.982\\
5\tabcellsep 98.795\tabcellsep 101.016\\
6\tabcellsep 99.568\tabcellsep 99.989\\
Mean\tabcellsep 99.3705\tabcellsep 100.5308\\
Std\tabcellsep 0.5297\tabcellsep 0.5213\\
Deviation\tabcellsep \tabcellsep \\
\multicolumn{3}{l}{e) Limit Of Detection (LOD) And Limit of Quantitation}\\
(LOQ)\tabcellsep \tabcellsep \end{longtable} \par
 
\caption{\label{tab_6}Table 7 :}\end{figure}
 			\footnote{Volume XII Issue II Version I © 2012 Global Journals Inc. (US) March} 		 		\backmatter   			 \par
The developed method was found to be simple, accurate, precise, reproducible and can be used for dissolution studies and routine quality control analysis of PH in bulk and in tablet form.\par
We are thankful to Padmashree (Mrs.) Fatma Rafiq Zakaria, Honorable Chairman of Maulana Azad Educational Trust, Dr. Rafiq Zakaria Campus, Aurangabad for providing the research facilities. We are also thankful to Aarti Drugs Ltd., Thane for providing the gift sample of Pioglitazone hydrochloride. 			  			  				\begin{bibitemlist}{1}
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\end{document}
