Spectrophotometric Determination of Tenatoprazole in Bulk Drug and Pharmaceutical Dosage Form

 

Kumaraswamy G.¹*, J.M. Rajendrakumar³, J.V.L.N. Sheshagirirao², U. Ashok Kumar¹ and    M. Arunadevi¹

¹Department of Pharmaceutical Analysis, Trinity College of Pharmaceutical sciences, Peddapalli, Karimnagar (Dist) - 505172.A.P INDIA.                ²Deptartment of Pharmacy, Andhra University, Visakhapatanam, A P       ³Holy Mary Institute of Technology and Science (HITS), Ghatkesar, Hyderabad.

*Corresponding Author E-mail: kumaraswamy.gandla@gmail.com

ABSTRACT:

A simple efficient, precise and accurate spectroscopic method has been developed and validated for quantitative estimation of Tenatoprazole in bulk drug and pharmaceutical dosage form. Tenatoprazole is dissolved in 0.1M Hydrochloric and the resulting solution was then scanned in the UV range (200-400nm) in a 10 mm quartz cell in a double beam UV spectrophotometer. The λmax of Tenatoprazole was found to be 314nm.The method obey’s Beers law in the concentration range from 3-18 µg/ml. The correlation coefficient was found to be 0.9997 (r²═ 0.9997). The LOD and LOQ were found to be 0.9882 and 1.9482mg/ ml respectively. The result of estimation of marketed tablet formulation (Allegro) was found to be 99.47% with their % RSD 0.4362.The accuracy of the method was determined by recovery studies. The percentage recovery was found to be 99.83%. The method was validated statistically as per ICH guidelines. The method showed good reproducibility and recovery with % RSD less than 2. So, the proposed method was found to be simple, specific, precise, accuracy, linear, and rugged. Hence it can be applied for routine analysis of Tenatoprazole in bulk drug and the Pharmaceutical formulations.

 

 

INTRODUCTION:

Tenatoprazole (TPZ) a new drug and it is used as Anti Ulcer agent^1-3 which is chemically is 3-methoxy-8-[(4-methoxy-3,5-dimethyl-pyridin-2yl)methyl sulfinyl] 2,7,9-triazabicyclo [4.3.0] nona-2,4,8,10-tetraene (Fig.1). The review of literature revealed that no methods were reported for the estimation of Tenatoprazole in bulk and tablet dosage form. TPZ is not official in any pharmacopoeia.

 

The mechanism action^4-5 of Tenatoprazole is a prodrug of the proton pump inhibitor (PPI) class, which is converted to the active sulphonamide or sulfenic acid by acid in the secretary canaliculus of the stimulated parietal cell of the stomach. This active species binds to luminally accessible cysteines of the gastric H⁺ K⁺ -ATPase resulting in disulfide formation and acid secretion inhibition. Tenatoprazole binds at the catalytic subunit of the gastric acid pump with a stoichiometry of 2.6 nmol mg^-1 of the enzyme in vitro. In vivo, maximum binding of Tenatoprazole was 2.9 nmol mg^-1 of the enzyme at 2 hours after IV administration.

 

The binding sites of Tenatoprazole were in the TM5/6 region at Cys813 and Cys822 as shown by tryptic and thermolysin digestion of the ATPase labelled by Tenatoprazole. Decay of Tenatoprazole binding on the gastric H⁺ K⁺ -ATPase consisted of two components. One was relatively fast, with a half-life 3.9 h due to reversal of binding at cysteine 813, and the other was a plateau phase corresponding to ATPase turnover reflecting binding at cysteine 822 that also results in sustained inhibition in the presence of reducing agents in vitro.

 

But there is no method was reported for the estimation of TPZ in bulk drug and in formulation. Hence the present work aims to develop simple, precise, accurate and validated spectroscopic method for the estimation of TPZ in bulk and in tablet dosage form by using simple UV method^6-7. Confirmation of the applicability of the developed method was validated according to the International Conference on Harmonization (ICH)^{8- 9} for the determination of TPZ in bulk and in tablet dosage form.

The aim of the present work was simple UV spectroscopic method for the determination of TPZ in bulk and in tablet dosage forms.

 

Fig.1 Chemical Structure of Tenatoprazole

 

MATERIALS:

Instruments¹⁰:

Shimadzu UV-1800 UV/VIS spectrophotometer with 1cm matched quartz cells was used for spectral and absorbance measurements.

 

Drug Material and Chemicals:

Tenatoprazole was obtained as a gift sample from Dr. Reddy’s Laboratories; (Hyderabad, India). Hydrochloric acid was purchased from Qualigens fine Chemicals Mumbai. The tablet formulation Allegro (Enteric coated tablets) containing 40mg of Tenatoprazole manufactured by SIDEM Pharmaceuticals (U.K) â.The tablet dosage form of TPZ for oral administration. The molecular weight is 346.40 for Tenatoprazole.

All the Chemicals and reagents used were of AR grade.

 

Preparation of Reagents¹¹

0.1M Hydrochloric acid

8.5 ml of hydrochloric acid was made up to 1000 ml with distilled water.

 

Simple UV Spectroscopic Method

This method is based on the measurements of absorbance of TPZ at its λ max was found to be 314nm. 0.1M HCl of analytical reagent grade was selected as solvent for developing spectral characteristics of drug. The selection was made after assessing the solubility of the drug in different solvents.

 

Stock and working standard solutions:

15mg of Tenatoprazole raw material was accurately weighed and transferred into the 100 ml volumetric flask dissolved and made up to 100 ml with 0.1M HCl. The solution was observed to contain 150 mg/ ml.

 

The above stock solution (0.5–3.0ml of 150μg/ ml) was transferred into six 100 ml volumetric flasks and made up to mark with 0.1M HCl. The absorbances of resulting solutions were measured at 314nm using 0.1M HCl as blank. Calibration curve was plotted by using concentration Vs absorbance. The curve was linear with the concentration range of 3-18 μg/ ml.

 

Application of the proposed procedure for the determination of dosage form:

Marketed tablet formulation Allegro was used for analysis. Twenty tablets were weighed and their average net weight was calculated. The tablet was made to a fine powder.

The powder equivalent to equivalent to 15 mg of Tenatoprazole was transferred into a 100 ml volumetric flask, added 0.1M HCl to dissolve and made up to the volume. Then the solution was sonicated for 10 minutes. After sonication the solution was centrifuged at 100 rpm for 15 minutes. The solution was filtered through Whatmann filter paper No.41. From the clear solution, 2 ml of the solution was transferred into a 25 ml standard flask and made up to the mark with 0.1M HCl to produce 12 mg/ ml concentration the absorbance measurements were made six times for the formulation at 314 nm. The amount of Tenatoprazole present in formulation was determined by using slope and intercept values from calibration graph.

 

Recovery studies:

The accuracy of the proposed methods was examined by determining the recovery of the drug by standard addition technique. To the preanalysed formulation a known amount of raw material added and it can be analyzed by proposed method 15 mg equivalent of Tenatoprazole formulation was taken into a series of three 100 ml standard flasks. To that 2, 4 and 6 mg/ ml of TPZ raw material Solutions were added in to series of standard flasks 1, 2 and 3, respectively. Dissolved with 0.1M HCl and made up to volume with 0.1M HCl .The solutions were sonicated for 10 minutes. After sonication, the solution was centrifuged at 100 rpm for 15 minutes. The solutions were filtered through Whatmann filter paper No. 41.  From each standard flask, 2 ml of the clear filtrate was transferred into a series of six 25 ml standard flasks and made up to volume with 0.1M HCl. The amount of drug recovered was calculated. Each concentration was repeated for three times.

 

Fig.2.Absorption Spectrum of Tenatoprazole in 0.1 M HCl.

 

Table 1: Optical characteristics

Parameters	Values
λmax(nm)	314 nm
Linearity range	3-18
Sandell’s sensitivity (µg/cm2/0.001 A.U)	0.014146E09
Correlation coefficient (r)	0.9993
Regression equation (y=mx+c)	Y=0.07069X-0.008643
Slope(m)	0.07069
Intercept(c)	0.008643
LOD (µg/ml)	0.988293622
LOQ (µg/ml)	1.94829157
Standard error of mean	0.006240421

Table 2: Assay of commercial formulation

S.NO	Label amount mg/tablet)	Amount found*(mg/tablet)	%Label Claim	SD	% RSD	SE
1	40	40.31	100.08	1.421	0.436	0.8366
2	40	39.86	99.32
3	40	39.84	99.75
4	40	41.03	100.93
5	40	39.79	99.35
6	40	41.86	100.87
			Avg.=99.47

* Mean + SD of six observations

 

 

Table 3: Intra-day and inter- day Precision of the method

Precision	Experimental concentration(mg/ml)	Recovery (%)	*RSD (%)
Intra- day (n=6)	12.09	99.91	0.97
Inter -day
1 Day (n=3)	12.23	99.86	0.73
2 Day (n=3)	12.15	99.47	0.85
3 Day(n=3)	12.93	98.95	0.91
Mean (n=9)	13.05	99.43	1.35

* RSD = Relative Standard Deviation

 

 

Table 4: Recovery studies

Amount Present (µg/ml)	Amount added* (µg/ml)	Amount found* (µg/ml)	Amount recovered* (µg/ml)	Average ± S.D	% RSD	S.E
12.03	2	14.01	99.05	0.4396	0.9286	0.3847
12.03	4	16.06	101.50
12.03	6	17.93	98.96

* Mean + SD of six observations

 

 

Fig.3     Calibration curve of Tenatoprazole

 

RESULTS AND DISCUSSION:

A simple, selective, accurate, precise spectroscopic method for the estimation of TPZ in bulk and tablet dosage form has been developed and validated (fig.2). The linearity range in the concentration range of 3-18 mg/ ml. The optical characteristics such as correlation co-efficient, slope, intercept, molar obsorptivity, LOD and LOQ were calculated and are shown in Table 1. The correlation co-efficient was found to be 0.9997. (r²═ 0.9997. shown in fig.3). It indicated that the concentrations of TPZ had good linearity. The LOD and LOQ were found to be 0.9882 and 1.9482mg/ ml respectively. The amount of TPZ in tablet formulation was found to be 99.47% shown in Table.02. To study the precision of the method the analysis of formulation was carried out for six times. The % RSD values were found to be 0.4410 by the low % RSD values, the precision of the methods were confirmed.  Further the precision was confirmed by intermediate precision. The analysis of formulation was carried out for three times in the same day and on three successive days. The % RSD values for interday and intraday analyses of formulation was found to be less than 2% and are shown in table 3. Further the precision of the method was confirmed by the repeatable analysis of formulation. The percentage recovery was found to be in the range of 98.96- 101.50%. The procedure was repeated for 3 times for each concentration. The % RSD was found to be 0.4362%.It indicated that the method has good precision. The low % RSD value indicated that there is no interference due to excipients used in formulation. Hence, the accuracy of the method was confirmed. The results of recovery studies are shown in table 4.

 

CONCLUSION:

The proposed method is simple, accurate, precise and selective for the estimation of Tenatoprazole in bulk and in tablet dosage forms. The method is economical, rapid and do not require any sophisticated instruments contrast to chromatographic method. Hence it can be effectively applied for the routine analysis of TPZ in bulk and in tablet dosage forms.

 

ACKNOWLEDGEMENTS:

The authors wish to thank Dr. Reddy’s Laboratories, Hyderabad for providing the gift sample of Tenatoprazole. The authors are thankful to Mr. D. Manohar Reddy, chairman, Trinity Educational Society and Dr. M. Arunadevi, Principal, Trinity College of Pharmacy for their kind help and providing all necessary facilities.

REFERENCE:

1.        Available from: http//www.en.wikipedia.org/wiki/Tenatoprazole.

2.        Available from: http//www.pubmed.pharmacol.com/Tenatoprazole.

3.        Available from: http//www.Rx list .com/Tenatoprazole.

4.        Shin JM, Choo YM, Sachs G. Aliment Pharmacol Ther., Blackwell Publishing Ltd, 2006, pp.2-8.

5.        Sharma, B.K. Instrumental Methods of Chemicals Analysis.Goel Publisher House, Meerut, 1994, 13^th ed: pp. 7.

6.        Mendham, J., Denny, R.C., Barnes, J.D. and Thomas, M.J.K. Vogel’s, Text book of Quantitative Chemical Analysis. Pearson Education Pvt. Ltd., New Delhi, 2002, 6^th ed: pp. 261-263, 268, 277, 653, 654.

7.        James, W. and Munson. Pharmaceutical Analysis Modern Methods Part B. International Medical Book Distributors, Mumbai, 2001, 16, 51, 76.

8.        Code Q2A, Text on Validation of Analytical Procedures. ICH Tripartite Guidelines, Geneva, Switzerland, 27 October, 1994, 1-5.

9.        Code Q2B, Validation of Analytical Procedures; Methodology. ICH Tripartite Guidelines, Geneva, Switzerland, and 6th November, 1996, 1 - 8.

10.     Anonymous. Shimadzu Instruction manual Pharamaspec UV-1800 series operation guide, Shimadzu Corporation, Kyoto, Japan, 2001, 6-2.

 

 

 

 

Received on 01.11.2010          Modified on 19.11.2010

Accepted on 03.12.2010         © RJPT All right reserved