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Validated Analytical Method Developed for Estimation of Ondansetron by Spectroscopy in Pharmaceutical Dosage Form
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Research - (2021) Volume 11, Issue 11

Validated Analytical Method Developed for Estimation of Ondansetron by Spectroscopy in Pharmaceutical Dosage Form

Sehgal Tanya*, Praveen K and Meenu C
*Correspondence: Sehgal Tanya, Department of Pharmacy, Shri Guru Ram Rai College of Pharmaceutical Sciences, Shri Guru Ram Rai University, Patel Nagar, Dehradun, Uttarakhand, India, Email:
Department of Pharmacy, Shri Guru Ram Rai College of Pharmaceutical Sciences, Shri Guru Ram Rai University, Patel Nagar, Dehradun, Uttarakhand, India

Received: 19-Nov-2021 Published: 17-Dec-2021

Abstract

The present research was undertaken to develop a spectroscopic method for determination of Ondansetron in pharmaceutical dosage forms. This paper describes a simple, rapid, accurate and precise UV-spectroscopic method for the assay of Ondansetron in bulk and marketed tablet dosage forms. The method is based on UV spectroscopic technique. Ondansetron shows the maximum absorbance at 302 nm in absorption maxima method. Drug followed the linearity in the range of 4-24 μg/ml for this method with correlation coefficient (r2) of 0.999. The results of analysis have been validated statistically and recovery studies confirmed the accuracy of the proposed method. The method was validated as per the International Conference on Harmonization (ICH) guidelines. The proposed method is recommended for routine analysis since it is rapid, simple, accurate and sensitive.

Keywords

Validation, Ondansetron, Ethanol, Precision.

Introduction

Ondansetron is chemically named as 9-methyl-3-[(2-methyl-1Himidazol- 1-yl) methyl]-2,3,4,9-tetrahydro-1H-carbazol-4-one, is official in IP, BP and USP. It is 5-HT3 receptor antagonist used mainly as an antiemetic (to treat nausea and vomiting). The antiemetic activity of the drug is brought about through the inhibition of 5-HT3 receptors present both centrally (medullary chemoreceptor zone) and peripherally (GI tract) [1,2]. Literature review shows that there are developed methods including spectroscopic, HPLC and HPTLC methods for the estimation of Ondansetron alone and in combination of other drugs like Omeprazole, Rabeprazole etc. There are developed Spectroscopic methods of analysis in single or in combination. Ondansetron shows absorption in UV-visible range in acidic media produced by hydrochloric acid was measured in absorption maxima method. Inthe present investigation simple and sensitive UV spectroscopic method have been developed for the quantitative estimation of Ondansetron in bulk and its marketed formulations with good accuracy and economy [3].

Materials and Methods

Active pharmaceutical ingredient used

Ondansetron drug from Sample from Nitin Pharmaceuticals Company is used along with Ondem drug from Alkem laboratories Ltd. The Instruments and chemicals used are shown in Table 1.

Table 1. Instruments and chemicals used.

S.No Name Model Supplier/manufacturer
Instruments
1 Single beam UV spectrometer Cary 60 UV Visible Agilent tech
2 Digital weight balance TX323L Shimadzu Instrument Pvt.Ltd
Chemicals
1 Ethanol   Central Drug House Pvt.Ltd

Method of development

Preparation of standard stock solution: Stock solutions of Ondansetron was prepared by transferring 50 mg of the drug in 50 ml volumetric flask and dissolved in 30 ml of ethanol and the volume was made up to the mark with ethanol. 2.5 ml of this solution was transferred to additional 25 ml volumetric flask and further diluted up to 25 ml mark with ethanol .This standard solution contained 100 μg of drug per ml.

Selection of wavelength maxima (λmax): Pipette out 1 ml of working standard solution and transfer into 10 ml volumetric flask and the volume was made up to the mark with solvent to get the concentration 10 μg/ml. The resulted 10 μg/ml solution was scanned in UVSpectrophotometer between 200-400 nm using ethanol as blank. The wavelength maxima were found at 302 nm [4].

Preparation of Calibration curve: Pipette out 0.4, 0.8, 1.2, 1.6, 2.0, and 2.4 working standard solution and was transferred into eight separate 10 ml volumetric flasks and the volume of all of them was made to 10 ml with ethanol to get the concentrations 4, 8, 12, 16, 20, 24 μg/ml respectively. Absorbance of the resultant solution was measured at 302nm using ethanol as blank. A graph was plotted between the concentrations and their respective absorbance. The response of the drug was found linear in the entire investigational range of 4-24 μg/ml [5]. The calibration equation was obtained y=0.0397x + 0.0008 with 0.999 correlation coefficient. The Beer’s-Lambert law was obeyed in the concentration range of (4-24 μg/ml) at 302 nm as shown in the Figure 1. Result was shown in Table 2.

pharmascholars-ondansetron

Figure 1. Calibration curve of ondansetron.

Table 2. Data for calibration curve of Ondansetron.

S. No. Concentration (μg/ml) Absorbance*
1 4 0.179366
2 8 0.3117
3 12 0.470466
4 16 0.6045
5 20 0.761133
6 24 0.9220333

Determination of optical parameters

The molar absorptivity and Sandell’s sensitivity were calculated as

Molecular absorptivity=AM/CT

A=Absorbance

M=Molecular weight

C=Concentration

B=Path length

Sandell’s sensitivity=M/E

M=Molecular weight

E=Molecular absorptivity

Other optical parameter that is beer’s limit, slope, intercept and correlation coefficient were calculated from calibration curve.

The table shows the optical and regression characteristics of ondansetron. This shows that the method is linear and obeys Beer’s law in the concentration range from 4-24 (μg/ml) with the molar absorptivity of 11617.169 litre mole-1cm-1 and sandell’s sensitivity of 25.255 × 10-3 μg/cm2/0.001 absorbance unit (Tables 3 and 4).

Table 3. Linearity, E1%1 CM, Absorptivity (L gm-1 cm-1), Molar Absorptivity (L mol-1 cm-1).

S. No Conc. (μg/ml)     Absorption    Mean E1% Absorptivity M.Absorptivity
       A1 A2 A3
1 4 0.1787 0.187 0.1727 0.1793 448.25 44.82 13150.188
2 8 0.3111 0.31 0.3145 0.3117 389.62 38.96 11430.846
3 12 0.4753 0.4628 0.4853 0.4744 395.33 39.533 11600.449
4 16 0.6067 0.6057 0.6011 0.6045 377.81 37.781 11084.945
5 20 0.7662 0.751 0.7662 0.7611 380.55 38.055 11165.337
6 24 0.924 0.9144 0.9277 0.922 384.16 38.416 11271.254

Table 4. Optical parameter of ondansetron.

Parameters Observation
Beer’s law limit Apr-32
Molar absorpivity 11617.169
Sandell’s sensitity (mg/cm/0.001 absorbance unit) 0.025255
Regression equation (y=mx+c)
m=slope, c=intercept
0.037
0.022
Correlation coefficient 0.999

Results and Discussion

Validation of proposed method according to ICH guidelines

Precision Repeatability: Pipette out 1.2 ml standard solution and was transfer into a series of nine 10 ml volumetric flasks. It was diluted to 10 ml with ethanol to get 12 μg/ml solutions [6,7]. Absorbance of the resultant solutions was measured at 302 nm using ethanol as blank. The result was obtained and summarized in the Table 5.

Table 5. Study of repeatability.

Conc. (μg/ml) Absorbance Observed Conc. (μg/ml) Mean Conc. (μg/ml) SD RSD
12 0.4656 12 12.08 0.00041 0.082258
0.4657 12
0.4679 12.1
0.4785 12.3
0.4623 11.9
0.4727 12.2

Keeping the concentration of the drug same procedure was repeated 6 times. The calculated RSD for repeatability study is 0.082258, which is acceptable; this shows a good repeatability of method.

Intra-day precision: Pipette out 0.8,1.2,1.6 ml working solution and was transferred into separate 10 ml volumetric flasks and the volume was made up to 10ml with ethanol to get the concentrations 8,12,16 μg/ml . Absorbance of the resultant solutions was measured at 302 nm using ethanol as blank. Such 6 revisions were performed within a day at 3 and 6 hrs interval. The result was summarized in the Table 6. The calculated mean RSD was 0.08683

Table 6. Study of intraday precision.

Conc.  (μg/ml) Absorbance Observed Conc. (μg/ml) Mean Conc. (μg/ml) SD % RSD
0 hr 3 hrs 6 hrs 0 hr 3 hrs 6 hrs
8 0.3451 0.3283 0.2808 8.4 8.2 7.2 7.93 0.000143 0.04288
12 0.4785 0.4727 0.453 12.3 12.2 11.6 12.03 0.000473 0.10136
16 0.6174 0.6092 0.5996 15.9 15.7 15.5 15.7 0.0007 0.11625

Inter-day precision: Pipette out 0.8,1.2,1.6 ml working solution and transfer into separate 10 ml volumetric flasks. Dilute all of them to 10 ml with ethanol to get solution of concentrations 8,12,16 μg/ml. Absorbance of the resultant solutions was measured at 302 nm using ethanol as blank. Such six studies were performed for day one day two day three intervals. The result was summarized in the Table 7.

Table 7. Study of Interday precision.

Recovery at Nominal Absorbance Observed Conc. ( μg/ml) % Recovery
80% 18=10+8 0.6981 17.8 98.88
80% 18=10+8 0.698 17.9 99.44
80% 18=10+8 0.6983 18 100
100% 20=10+10 0.7326 19.6 98
100% 20=10+10 0.7908 20 100
100% 20=10+10 0.7461 19.8 99
120% 22=10+12 0.8478 22 100
120% 22=10+12 0.8424 21.8 99.09
120% 22=10+12 0..8475 21.9 99.54

Accuracy: Pipette out 1 ml standard solution and transfer into 10 ml volumetric flasks. Nine such transfers were made. Spike three of volumetric flask with the solutions with 0.8 ml of working solution (Prepared from Formulation) and dilute each to 10 ml with ethanol to get 18 μg/ml solutions. Spike another three of the solutions with 1 ml of working solution and dilute each to 10 ml with ethanol to get 20 μg/ ml solutions. Spike last three of the solutions with 1.2 ml of working solution and dilute each to 10 ml with ethanol to get 22 μg/ml solutions. Absorbance of the resultant solutions was measured at 302 nm using ethanol as blank. The obtained results were summarized in the Table 8.

Table 8. Study of accuracy.

Nominal conc. Without excipients With excipients % Interference
  Absorbance Observed Conc. (μg/ml) Absorbance Observed Conc. ( μg/ml)
12 0.4755 12.1 0.476 12.2 1.01
12 0.4458 11.5 0.4465 11.9 0.99
12 0.4753 12 0.5986 15.4 1.28
12 0.4555 11.7 0.4727 12.1 1
12 0.4623 11.9 0.4675 12 1
12 0.4785 12.3 0.469 11.7 0.97

Specificity: Specificity study was carried out by observing any interference in absorbance of drug in the presence of common excipients like starch, talc, lactose, magnesium stearate etc. Absorbance of 10 μg/ ml drug solution with and without excipients was measured at 302 nm using ethanol as blank [8]. The results obtained were summarized in the Table 9.

Estimation of ondansetron in pharmaceutical dosage form (Ondem: 4 mg)

20 tablets were weighed and the average weights of the tablets were calculated. The tablets were powdered and weighed accurately [9]. A quantity of powdered containing about 20 mg of ondansetron was transfer into 50 ml volumetric flask and 15 ml ethanol was added. Sonicated for 15 minutes and the volume were made upto 50 ml with solvent then was mixed and filtered. 2.5 ml of the filtrate was taken and was made up to 25 ml with ethanol. Further 1.6 ml of the resulting solution was diluted to 10 ml with ethanol. The absorbance of this resulting solution was measured at 302 nm. The above procedure was repeated for three times. The result obtained is summarized in the Tables 10 and 11.

Table 10: %Assay of ondansetron in pharmaceutical dosage form (ondem, 4 mg).

Sr. No. Absorbance Conc. (μg/ml) Dil. Factor Weight taken (mg) Avg. weight (mg) Label claim (mg) Assay%
1 0.6765 15.9 2500 40 40 4 99.37
2 0.6514 16.1 2500 40 40 4
3 0.6846 16.2 2500 40 40 4
4 0.6297 15.8 2500 40 40 4
5 0.6247 16.1 2500 40 40 4

 

Table 11. Summary of validation parameters of ondansetron.

Validation parameters Observation
Linearity and range 4-24
Correlation coefficient (f) 0.999
Precision (RSD)
  • Repeatibility
0.082258
  • Intraday
0.08683
  • Interday
0.06222
%Recovery 99.327
%Interference 1.2
%Assay 99.37

Conclusion

A simple and sensitive spectroscopic method for quantitative determination of ondansetron in either pure form or in pharmaceutical dosage for was developed. Ondansetron showed maximum absorbance at 302nm in solvent. It has linear response in the entire range of 4to 24 μg/ml with correlation coefficient of 0.999, with molar absorptivity of 11582.332 litre mole-1cm-1 and Sandell’s sensitivity of 0.025255 microgram/cm2/0.001 absorbance units. The linear regression equation obtained is y=0.037x+0.022. The method has good precision <2% and accuracy is 99.37 ± 0.50. No significant interference was observed in the absorbance of the drug in the presence of common excipients. The method was statistically validated according to ICH.

The method was employed for the quantitative determination of tablet dosage form. In conclusion, the developed spectroscopic methods are simple, accurate, and reproducible, and can be used in routine analysis of Ondansetron in bulk.

Acknowledgement

The authors are grateful to Nitin pharmaceuticals, Karnal for providing Ondansetron. The authors are also highly grateful to Shri Guru Ram Rai College of Pharmaceutical Sciences Dehradun for providing all the laboratory facilities to carry out the work.

References

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