Int J Curr Pharm Res, Vol 12, Issue 5, 59-62Original Article


DEVELOPMENT AND VALIDATION OF UV-SPECTROSCOPIC METHOD FOR ESTIMATION OF TIOTROPIUM BROMIDE IN API AND PHARMACEUTICAL DOSAGE FORM

ANJALI P. KOKANE*, BHAGYASHRI S. SHINDE, M. S. KALSHETTI

Department of Pharmaceutical Quality Assurance, D. S. T. S Mandal’s College of Pharmacy, Solapur, Maharashtra, India
Email: anjalikokane333@gmail.com

Received: 18 Jun 2020, Revised and Accepted: 19 Aug 2020

ABSTRACT

Objective: The current work intended towards the developed and validated UV spectrophotometric method for the estimation of Tiotropium Bromide in API and pharmaceutical formulation.

Methods: The UV spectrum of tiotropium bromide in methanol shows maximum absorbance at 230 nm. Calibration curve method was used for the estimation of tiotropium bromide in API and pharmaceutical formulation.

Results: Maximum absorbance obtained in 230 nm. Calibration curve plotted in concentration range 20-120 µm/ml exhibit the linearity relationship with line equation y=0.0057x+0.3319. The Accuracy was found to be 80-100.5%. The precision %RSD=0.1110, and the limit of detection(LOD)=4.12and limit of quantification (LOQ= 12.50. The method was found to comply all the validation parameters as per the ICH guideline in indicating the sensitivity of the method analyte.

Conclusion: This method is used as satisfactory for the routine analysis of tiotropium bromide in API and pharmaceutical dosage forms.

Keywords: Tiotropium bromide, UV Spectrophotometer, Methanol, Validation

© 2020 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
DOI: http://dx.doi.org/10.22159/ijcpr.2020v12i5.39767. Journal homepage: https://innovareacademics.in/journals/index.php/ijcpr

INTRODUCTION

The Tiotropium Bromide chemically is (12, 4β, 5∝, 7β,)-7-[(Hydroxydi-2-thieyl-acetyl) oxy]-9, 9-dimethyl-3-oxa-9-azoniatricyclo [3.3.1.0] [1]. Tiotropium is used as a maintenance treatment of chronic obstructive pulmonary disease (COPD). It may also be used as an add-on-therapy in people with moderate-to-severe asthma on medium to high dose inhaled corticosteroid (ICS) [2, 3] Tiotropium bromide smooth muscles relaxation and bronchodilation [4, 5]. A tiotropium metered inhalation spray is indicated for the maintenance of bronchospasms in COPD to prevent exacerbation of COPD and to treat asthma in patient six or more year old [6, 7]. It is used by inhalation through the mouth [8, 9]. Onset typically begins within half an hour and lasts for 24 h. It is not, however, approved for acute exacerbation of COPD or acute worsening asthma [10, 11].

Fig. 1: Chemical structure of tiotropium bromide [1]

MATERIALS AND METHODS

Selection of solvent and instrument

Tiotropium bromide is dissolving in various solvents for trails of selecting the ideal solvent [12, 13]. Various solvents i.e. methanol, water, ethanol, acetonitrile. Tiotropium bromide is soluble in methanol, and better absorption was found to be at230 nm. UV-Visible Spectrophotometer (shimadzu AY220) [14-16]. Reagents and Materials is API-Tiotropium Bromide was obtained as a gift sample from the Vamsi Pharmaceutical Ltd. Solapur, Maharashtra. Capsule is purchased from the local pharmacy in Solapur. Brand name is Tiova (Cipla); methanol was used for this study.

Experimental work

Method development

Selection of detection wavelength

Dilution of Tiotropium bromide was prepared from the stock solution (50µg). Tiotropium bromide was scanned over a range of 200-400 nm. Drug showed maximum absorbance at 230 nm was selected as the wavelength for detection.

Preparation of standard drug solution

10 mg of Tiotropium bromide was weighed and transferred into a 10 ml volumetric flask containing methanol. Concentration of stock solution is (1000µg/ml). Then pipette out 2 ml form the stock solution and adjust to volume.

RESULTS AND DISCUSSION

Method validation

The method was validated for several parameters like a preliminary analysis of drug Linearity, Accuracy, Precision, Robustness, Ruggedness, Limit of Detection, Limit of Quantification and specificity of tiotropium bromide.

Preliminary analysis of drug

Observation and result of preliminary analysis of Tiotropium bromide.

Preliminary analysis of drug

Test Observation Result
Description White powder Complies
Solubility Soluble in methanol and acetonitrile Complies

The UV-Spectrum of Tiotropium bromide in methanol showed at max absorbance at 230 nm.

Fig. 2: UV visible spectrophotometer graph of tiotropium bromide

Fig. 3: Calibration curve for tiotropium bromide

Linearity and range

It is a linear relation between absorbance and concentration of drug was evaluated using three replicates over concentration range in 20-120(µg/ml) by making the replicates (fig. 3)

The wavelength for linearity was scanned at 230 nm. By taking five different concentrations for linearity the regression coefficient was found to 0.999 i.e. in the limit of standard. Hence linearity was found to be validated.

Accuracy

This study was carried out at three different levels that are 80%, 100%, 120%, by standard addition method. Analyzed sample by triplicate by according to the method. Known amount of standard tiotropium bromide was spike on the capsule sample. Check the absorbance and calculated (table 1)

Precision

Precision is defined as an analytical procedure is define the closeness of agreement between samples of measurements obtained from multiple sampling of the same homogenous sampling in specific conditions. It is determined by inter-day. Reading is taking 3 times on the same day. The percent relative standard deviation. (%RSD) was calculated (table 2)

Robustness

Robustness is a measure of its capacity of analytical procedure to remain unaffected by small changes or this method small deliberate variation in the method parameters. Main aim of this test is to create a method that allow for some variations in the parameters. In this study wavelength was change at±5 nm. Then robustness is calculated.

Table 1: Result of accuracy

S. No. % level Spike amount (µg/ml) Spiked amount (wrt sample) Abs. Amount recovered %RSD % recovery
1 80 79.440 79.4 0.829 80.651 0.9
2 100 99.300 99.3 1.051 100.550 0.3
3 120 119.160 119.2 1.227 100.5 1.5

Table 2: Result of intra-day morning precision

S. No. Concentration (µg/ml) Absorbance SD %RSD
1 60 0.678
2 60 0.679
3 60 0.679
4 60 0.678 0.000753 0.111028
5 60 0.677
6 60 0.678

Table 3: Result of robustness

S. No. Wavelength Absorbance SD %RSD
1 230 0.444 0.006083 1.3919
0.434
0.433
Avg= 0.43
2 232 0.432 0.005859 1.3428
0.434
0.443
Avg= 0.436

By change in wavelength i.e. 232 nm. % RSD is less than 2% i.e. within the range. So parameter was validated (table 3)

Ruggedness

Ruggedness is defined as, reproducibility of the results when the defined method was performed under different analysts, laboratories, columns, chemicals, solvents, instruments, sources of reagents and etc.

Limit of detection

Detection limit is defined as, the lowest amount of analyte in a sample can be detected (table 6)

LOD= 3.3(SD/S) SD= Standard deviation, S= slope of the curve

Table 4: Result of ruggedness

Concentration (µg/ml) Absorbance Statistical analysis
60 0.678 Avg =0.678
60 0.679 SD =0.001
60 0.677 % RSD =0.147

Table 5: Result of ruggedness

Concentration (µg/ml) Absorbance Statistical analysis
60 0.679 Avg =0.678
60 0.678 SD =0.001
60 0.677 % RSD =0.147

By change in analyst and laboratory, there is no effect on absorbance with same conditions; hence, parameter was validated (table 4 and 5).

Table 6: Result of limit of detection

LOD(µg/ml) 4.12

Table 7: Result of limit of quantification

LOQ (µg/ml) 12.50

Limit of quantitation [17]

Limit of quantification is defined as; it is an individual analytical procedure is the lowest amount of an analyte in the sample, which can be an exact value (table 7)

LOQ=10(SD/S) SD= Standard deviation, S= slope of the curve

CONCLUSION

The proposed method is found to be accurate, precise, stable, linear, specific, and simple for quantitative estimation of tiotropium bromide in the bulk and pharmaceutical dosage form. Hence the present method is suitable for the routine assay of tiotropium bromide in bulk and pharmaceutical formulation.

ACKNOWLEDGEMENT

Authors are thankful to the D. S. T. S. Mandal’s College of Pharmacy, Solapur. Maharashtra India for providing the facility for the conduct of the research work. Authors are also thankful to Prof. Dr. M. S. Kalshetti for their consistent support and assistance throughout the project work.

FUNDING

Nil

AUTHORS CONTRIBUTIONS

All the authors have contributed equally.

CONFLICT OF INTERESTS

Declared none

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