Full HTML 12 V1 I2

ANALYTICAL METHOD DEVELOPMENT AND ITS VALIDATION FOR ESTIMATION OF PANTOPRAZOLE SODIUM IN BULK AND TABLET DOSAGE FORM BY UV- SPECTROSCOPIC METHOD


A. Phadtare*, S. C. Daswadkar, M. A. Kuchekar
Dept. of Pharmaceutical Chemistry, Padmashree Dr. D. Y. Patil College of Pharmacy, Akurdi, Pune, India

ABSTRACT 

A simple, sensitive, spectrophotometric method in UV region has been developed for the determination of Pantoprazole in bulk and tablet dosage form. The method have been developed and validated for the assay of Paracetamol using Methanol and water as diluents.  Solution of Pantoprazole in Methanol shows maximum absorbance at 267 nm in zero order spectrum method (method A), in first order derivative spectra (method B) show sharp peak at 258 nm when n=1, and in method C calculation of area under curve (AUC) for analysis of Pantoprazole in wavelength range 262-272nm. The drug followed the Beer-Lambert’s law in the concentration range of 2-10 µg/ml. Result of analysis were validated statically and were found to be satisfactory. All the parameters of the analysis were chosen according to ICH [Q2 (R1)] guideline.

Keywords: Pantoprazole sodium sesquihydrate, Zero order, First order, Area under order (AUC).

INTRODUCTION 

Pantoprazole is used as an Anti ulcerative agent in the form of Pantoprazole sodium sesquihydrate   (RS)-6-(difluoromethoxy)-2-[(3, 4-dimethoxypyridin-2-yl) methylsulfinyl]-1H-benzo[d]imidazole.1  Pantoprazole sodium is officially listed in I.P.2010 2 and B.P.2010. 3It is a proton pump inhibitor. PPS binds irreversibly to H+ K+ ATPases and suppresses the secretion of acid. PPS is pharmaceutically formulated as gastro-resistant tablets containing 20.0, 40.0 mg Pantoprazole. The methods reported for quantitative determination of Pantoprazole in bulk or pharmaceutical formulations include titrimetry, colorimetery and high performance liquid chromatography.4-10 This paper presents the simple, accurate and reproducible UV spectrophotometric methods for determination of Pantoprazole in tablet dosage form. In the literature survey it is found that methods have been reported for estimation of Pantoprazole in tablet dosage form by UV Spectrophotometry.11-15 

MATERIALS AND METHODS

Instrument

A Shimazdu UV/VIS Spectrophotometer was used with 1cm matched quartz cells and spectral bandwidth of 2 cm.  

Materials

Standard gift sample of Pantoprazole (RS)-6-(difluoromethoxy)-2-[(3, 4-dimethoxypyridin-2-yl) methylsulfinyl]-1H-benzo[d]imidazole, was procured from Litaka Pharmaceuticals Ltd. Talegaon Dabahade

Solvent used: Methanol, Distilled Water

Stock Solution Preparation: Weigh 10 mg of drug dissolve in 10 ml methanol Keep it 15-20min in ultrasonicator. Final volume make up with distilled water (100μg/ml).

Method A

From the stock solution of Pantoprazole appropriate volumes were pipette out and transferred to 10ml volumetric flasks. Keep it 15-20min ultrasonicator. The volume was made up to the mark with distilled water to give the samples of desired concentrations like 2, 4, 6, 8, 10mg/ ml. The absorbance maxima of these solutions were found at 267 nm. (Fig.1) a linear graph of absorbance vs. concentration was obtained. The concentration range over which the drugs obeyed Beer-Lambert’s law was found to be 2-10mg/ml for Pantoprazole. The concentration of sample solution was determined from calibration curve.

Figure 1: zero order spectrum 

Method B

The first order derivative spectra at n=1 (method b) showed a sharp peak at 258nm (figure 2). The absorbance difference at n=1 is calculated by the inbuilt software of the instruments which was directly proportional to the concentration of the standard solution .The standard drug solution was diluted so as to get the final concentration in the range of 2-10µg/ml and scanned in the first order derivative spectra. The calibration curve of Pantoprazole absorbance difference against concentration of the drug showed linearity.

Figure 2: First order spectrum 

Method C

The AUC (Area under curve) method involves the Suitable concentrations of solutions were prepared accurately to determine the range of Pantoprazole for analysis. The standard solutions were scanned in the spectrum mode of the instrument from 400 nm to 200nm. The absorbance maxima of these solutions were obtained at wavelength 270nm. The area under the curve between 262nm to 272 nm was selected (Fig.3) for the calculation because the linearity was obtained within these areas with good reproducibility of results.

Figure 3: wavelength range selected for AUC

Analysis of tablet formulation

For estimation of pantoprazole in tablet formulation by three methods, twenty tablets were weighed. The tablet content was weighted and triturated to fine powder. Tablet powder equivalent to 10 mg. of pantoprazole was weighed and transferred to 100 ml volumetric flask and dissolved in 10 ml of methanol. It was kept for ultrasonification for 30 min. finally the volume was made up to the mark with distilled water. further pipette out 1ml from above solution and dilute up to 10 ml with distilled water this was then filtered through whatman filter paper no. 41 to get tablet stock solution of concentration of 10 μg/ml. various dilution of the tablet solution were prepared and analyzed for six times and concentration was calculated by using the calibration curve for three method.

All these method were validated according to ICH guidelines by carrying out analysis of six replicate samples of the tablets (table 2), Recovery studies were carried out at two different level i.e 80%100%, 120%by adding the pure drug (8, 10, and 12 mg respectively) to previously Analysed tablet powder sample from the amount of drug found percentage recovery was calculated (Table3).

RESULT AND DISCUSSION

The Methods A, B, C for estimation of Pantoprazole in tablet dosage form were found to be accurate and reproducible .beer- lambert’s was obeyed in the concentration range of 2-10ug/ml. in all these methods .The values of standard deviation were satisfactory and the recovery studies were close to 100%.hence These method can be useful in the routine analysis of Pantoprazole in bulk drug and formulation.  

Table 1: Optical Calibration Curve of Pantoprazole

Parameters Method A Method B Method C
max (nm)/wavelength range(nm)

Beer’s – Lambert’s range(µg/ml)

Coefficient of correlation (r2 )

Regression equation : Y = mx + c

a – Slope (m)

b – Intercept (c)

LOD

LOQ

Molar absorptivity

267nm

2-10µg/ml

0.99973

0.0542x+0.0006

0.0570

0.0

0.0008

0.0026

8.8×10-3

258

2-10µg/ml

1.000

0.001x+0.001

0.001

0.0

3.3

10

0.16×10-3

262-272nm

2-10 µg/ml

0.9997

0.0031x+0.0005

0.0031

0.0

0.0006

0.002

4.4×10-3

 Table 2: Estimation of Pantoprazole formulation tablet 

Method Tablet Formulation Label claim Amount found % mean S.D C.O.V S.E
A T1 40 39.78 99.98 0.1506 0.0159 0.0614
B T1 40 39.55 100.08 0.836 0.679 0.343
C T1 40 39.80 99.96 0.4527 0.4536 0.1848

 Table 3: Recovery Study Data 

Method Tablet Formulation Level Of

% Recovery

Label Claim Amount of std drug added %*     Mean %* Mean S.D C.O.V S.E
A T1 80 10 8 18.04 99.98 0.4055 0.4058 0.2314
100 10 10 20.04 100 0.1323 0.1317 0.0763
120 10 12 22.01 100.05 0.2946 0.2937 0.1701
B T1 80 10 8 18.06 99.98 0.6892 0.6896 0.3979
100 10 10 20.05 100 0.6759 0.6774 0.3902
120 10 12 22.01 99.96 0.5128 0.5103 0.2960
C T1 80 10 8 17.75 99.50 0.6971 0.6990 0.4025
100 10 10 20.03 100.03 0.2611 0.2616 0.1508
120 10 12 22.03 100.03 0.6312 0.6315 0.3644

 REFERENCES

  • The Merck Index – an encyclopaedia of chemicals, drugs and biologicals, 14th edition, (7014), 1209.
  • Indian Pharmacopoeia, 6th Edition, Ministry of Health and Family welfare, published by The Indian Pharmacopoeia Commission, Ghaziabad, vol-II, 2010, 1856-57.
  • British Pharmacopoeia. 6th Edition (6.1-6.5), vol-II, (2010);
  • Kumar R., Singh P. and Singh H., Development and validation of RP-HPLC method for simultaneous estimation of Naproxen and Pantoprazole in Pharmaceutical dosage form, International Journal of Pharmaceutical Research and Development. 2011; 12: 12-14
  • Sivakumar T. , Manavalan R. and Valliappan K., Development and Validation of a Reversed-phase HPLC method for simultaneous determination of Domperidone and Pantoprazole in pharmaceutical dosage forms, ACTA Chromatographica, 2007; 18:30-32.
  • Patel G. H., Prajapati S.T. and Patel C.N., HPTLC Method Development and Validation for simultaneous Determination of Cinitapride and Pantoprazole in Capsule Dosage Form, Res. J. of Pharm. and Tech. 2011; 4(9):1428-1431.
  • Birajdar A.S. , Meyyanathan S.N., Bojraj S., Application of UV-Spectrophotometry and RP-HPLC for Simultaneous Determination of Rabrprazole (Internal standard Pantoprazole) and Domperidone in Pharmaceutical Dosage Form.” Pelagia Res. Lib. Der Pharmacia Sinica. 2010; 1(3): 69-78.
  • Siddartha B.and I. Sudheer Babu, Analytical method development and method validation for the estimation of Pantoprazole in tablet dosage form by RP-HPLC , Scholars Research Library Der Pharma Chemica, 2013; 5 (4):99-104
  • Bansode P. S., Kamble R. K., Chauhan C. S. and Bansode S. S., Method development and validation of related substances in Pantoprazole Sodium by RP HPLC, Journal of Chemical and Pharmaceutical Research, 2014; 6(11): 942-948
  • Hegazy M, Yehia A, Mostafa A, Stability indicating chromatographic methods for simultaneous determination of mosapride and Pantoprazole in pharmaceutical dosage form and plasma samples, Chromatographica, 2011; 74: 839-845.
  • Jigar Pandya , Mr. Sagar Solanki, Dr. Mandev Patel Development and Validation of Differential Spectrophotometric method for Determination of Pantoprazole in Tablet Dosage Form Journal of pharmaceutical science and bioscietific Research,2012; 2(1):2-4
  • Rajnish K., Harinder S. and Pinderjit S., Development of UV Spectrophotometric method for estimation of Pantoprazole in pharmaceutical dosage forms, Journal of Chemical and Pharmaceutical Research, 2011; 3(2):113-117.
  • Ravi Kumar P. ,Bhanu Prakash P., Murali Krishna M., Santha Yadav M and Asha C., Simultaneous Estimation of Domperidone and Pantoprazole in Solid Dosage Form by UV Spectrophotometry, E-Journal of Chemistry, July 2006; 3(12):142-145.
  • Mohamed M. Barak, Mohamed E. El-Sadek, Lobna M. Abdel-Aziz, Samar S. El-Bermawi, Validated spectrophotometric determination of Pantoprazole sodium in tablets using 2, 4-Dinitrofluorobenzene through nucleophilic substitution reaction, Asian Journal of Pharmaceutical Analysis And Medicinal Chemistry,2014; 2(1):26 – 36.
  • Jasmine Karanjia , Development and Validation of Chemometric Assisted Spectrophotometric Technique for Simultaneous Estimation of Cinitapride and Pantoprazole from Bulk and Combined Dosage Form, International Journal of Pharmaceutical Science and Research 2015; 7(2):198-204