Developed a Validated Analytical Procedure for Estimation of Ondansetron by UV-Spectroscopy as API and in tablets

 

Meenu C.¹, Pradeep K. Y.², Praveen K.³*

¹School of Pharmaceutical Sciences, SGRR University, Patel Nagar, Dehradun- 248001, Uttarakhand, India.

²Department of Pharmacy, LLRM Medical College, Meerut, Uttar Pradesh, India.

³Quantum School of Health Sciences, Quantum University, Mandawar, Roorkee- 247167, Uttarakhand, India.

 

ABSTRACT:

Pharmaceutical analysis and need of drug Analysis pharmaceutical analysis is an important part of pharmaceutical chemistry which involves the science of extraction. Analytical research and development is a process which assures quality attributes and quality control of the products. A modest specific, perfect and delicate Ultra Violet-Spectroscopyprocessdeveloped for the determination of Ondansetron in bulk as well as tablets. Ondansetron shows the maximum absorbance at wavelength 302nm. Ondansetron showed the linearity, range 04-24µg per ml for this procedure with Correlation Coeff. (R²) was 0.9991. The technique was found definite as no intervention was detected with excipients. The recovery studies confirmed the correctness of the anticipatedtechnique and outcome was found 99.32 with RSD ±0.66. The process was confirmed as per the ICH guidelines. The proposed analytical process is suggested for monotonous analysis of Ondansetron as API and in tablet unit dosage forms.

 

 

 

 

 

INTRODUCTION:

Ondansetron used to be established around 1984 by means of researchers working at Glaxo's laboratories in London. It is chemically 1, 2, 3, 4-tetrahydro-9-methyl-3-(2-methylimidazol-1-ylmethyl) carbazol-4-one. Ondansetron is the imidazole and carbazole family members of heterocyclic compounds¹. Ondansetron is 5-HT³ receptor antagonist and used in greatest cases as an antiemetic².

 

The antiemetic activity of the Ondansetron is introduced about due to the inhibition of 5-HT³ receptors contemporary both centrally (Medullary Chemoreceptor Zone) and peripherally (Gastrointestinal tract)². Some derivative UV spectophotometric techniques had been stated for Ondansetron in therapeutic dosage forms^3-7. HPLC analytical procedure for quantifiable purpose of Ondansetron in amalgamation had been stated in literature^8-13. HPTLC¹⁴ and LCMS^15,16 analytical methods have been also said for the valuation of Ondansetron had developed. Nonetheless due to the interferingtriggered by the preparation excipients the processes were not produce the acceptable results. Analytical approaches for the quantization of the Ondansetron in biological fluids may not be accessible. The unbiased of work was to improvetechnique which is accurate, precise and sensitive analytical method and its validation which could be further used for determination of Ondansetron during routine Pilot manufacturing, as API and in tablet unit dosage forms.

 

MATERIALS AND METHODS:

Materials:

RS of Ondansetron Active Pharmaceutical Ingredient was provided as gift tester by Nitin Pharmaceuticals, Ondem (Marketed formulation) by Alkem laboratories Ltd., and Ethanol from Central Drug House Pvt. Ltd.

 

Instruments:

The Ultra Violet-Spectroscopyquantities were conceded out with a Cary 60Single Beam UV spectrometer manufacturer by Agilent Tech. Digital Weight Balance: TX323L, Shimadzu was used.

 

Preparation of Standard Solutions:

Stock solutions of was set by transferred 50mg of the Ondansetron with 30ml of ethanol and diluted 50ml with ethanol in analytical Flask. The 2.5ml of previously set solution was diluted with 25ml ethanol. This standard sollutioncontained 100μg of drug per ml.

 

Selection of wavelength maxima (λmax):

Pipetted out 1ml of standard solution and diluted with 10 ml of ethanol to get the concentration 10μg per ml. The stemmedsolution was scanned in UVwith 200-400 nm wavelength used ethanol as a blank. The result was shown (Fig. 1).

 

Preparation of Calibration curve:

Pipetted out 0.4, 0.8, 1.2, 1.6, 2.0, and 2.4ml standard solutions and shifted into six separate 10ml analytical flasks and volume of all of them was made up to the mark 10ml with ethanol to get the concentrations 4, 8, 12, 16, 20, 24μg per ml respectively. Absorbance of the resultant solution was measured at 302nm ethanol used as a blank. A graph was plotted between the concentrations and their respective absorbance. The results were shown in (Fig. 2) and (Table 1).

 

 

Fig. 1: UV Spectra of Ondansetron.

 

 

Fig. 2: Calibration curve of Ondansetron

 

Validation of proposed method according to ICH guidelines¹⁷:

Repeatability:

Pipetted outs 1.20ml of standard solution shifted into a series of nine 10ml analytical flasksand diluted with ethanol to get the concentration of 12μg per ml. Optical density of the resultant solutions was dignified at 302nm ethanol used as a blank. The results were obtained and concise in the (Table 2).

 

 

 

Table 1: Linearity, Range, E^1%_1CM, Absorptivity (L gm-¹ cm-¹), and Molar Absorptivity (L mol-¹ cm-¹)

S. No	Concentration (μg per ml)	Absorption			Mean	E1%	Absorptivity	Molar Absorptivity
		A1	A2	A3
1	4	0.1787	0.1870	0.1727	0.1793	448.25	44.82	13150.188
2	8	0.3111	0.3100	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	0.7510	0.7662	0.7611	380.55	38.055	11165.337
6	24	0.9240	0.9144	0.9277	0.9220	384.16	38.416	11271.254
Mean: 11617.169

Table 2: Study of Repeatability

 

 

 

Intra-Day Precision:

Pipette outs 0.8, 1.2, 1.6ml working solutions weretransferred into separate 10ml analytical flasks and diluted with ethanol to get the concentrations 8, 12, 16μg/ml. Absorbance of the subsequent solutions was measured at 302nm ethanol used as a blank. Such sixrepetitions were performed within a day at 0, 3 and 6 hrs interval. The result was summarized in the (Table 3). The % RSD was less than 2%.

 

Inter-Day Precision:

Pipette outs 0.8, 1.2, 1.6ml working solutions were transferred into separate 10 ml analytical flasks and diluted with ethanol to get the concentrations 8, 12, 16μg/ml. Absorbance of the subsequent solutions was measured at 302nm. Such six studies were performed for day one day two day three intervals. The result was summarized in the (Table 4). The %RSD was less than 2%.

 

 

 

 

Accuracy:

Pipette out 1.00ml of standard solution was shifted into 10ml analytical flask. Nine such transfers were made. Spike three of volumetric flask with the solutions with 0.8ml of working solution (Prepared from Formulation) and diluted each with ethanol to get 18μg/ml solutions. Spike another three of the solutions with 1ml of working solution and diluted each with ethanol to get 20μg/ml solutions. Spike last three of the solutions with 1.2ml of working solution and diluted each with ethanol to get 22μg/ml solutions. Absorbance of the resulting solutions was determined. The gottenoutcomes were concise in the (Table 5).

 

Specificity:

Specificity study was carried out by observing any interference in absorbance of drug in the existence of conjoint excipients like Starch, Talc, Lactose, Magnesium Stearate etc. Absorbance of 10μg/ml drug solution with and without excipients was measured at 302nm. The results obtained were summarized in the (Table 6).

 

Table 3: 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	0.4530	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	0.00070	0.11625
Mean: 0.08683

 

Table 4: Study of Interday precision

Conc. (μg/ml)	Absorbance			Observed Conc.(μg/ml)			Mean\Conc(μg/ml)	SD	%RSD
	0 hr	24 hrs	28 hrs	0 hr	24 hrs	48 hrs
8	0.3333	0.2856	0.2983	8.5	7.8	7.6	7.96	0.00017	0.0552
12	0.4976	0.5141	0.4351	12.8	13.2	11.6	12.53	0.00037	0.0762
16	0.6297	0.6247	0.6057	16.3	16.1	16.0	16.13	0.00242	0.3862
Mean: 0.0622

 

Table 5: Accuracy

 

Table 6: Study of specificity

 

Table 7: % Assay of ondansetron in pharmaceutical dosage form (ondem, 4mg)

 

 

 

% Assay of Ondansetron tablets (Ondem, 4mg tab.):

Taken 20 tablets and weighed accurately for calculated the average weight. The tablets were powdered and aamount of powdered containing about 40 miligram of Ondansetronwas transferred into 50ml analytical flask with 15ml of ethanol. Sonicated this solution for 15 minutes and diluted with solvent up to 50ml then mixed and filtered. 2.5ml of the filtrate was taken and diluted up to 25ml with ethanol. Further 1.6ml of the resultant solution was diluted to 10ml with ethanol. The absorbance of this resultant solution was estimated at 302nm. The above procedure was repeated for three times. The results gottenwereconcise in the (Table 7).

 

RESULT:

The wavelength maximum of Ondansetron was found to be 302nm. The reply of the Ondansetron was found linear in the entire investigational range of 4 - 24μg/ml. The calibration equation was obtained y = 0.0397x + 0.0008 with 0.999 correlation coefficient. The Beer’s-Lambert law was conformedlinearity with 4-24μg per ml strength. The Molar absorptivity and Sandell’s sensitivity of Ondansetron were 11617.169 litre mole^-1 cm^-1, 25.255x 10^-3μg/cm²/0.001 respectively. The calculated RSD for repeatability study is 0.082258, which is acceptable; this shows a good repeatability of method. The method was very specific their no interference found. The accuracy was 99.88% RSD less than 2% respectively. The assay of drug in pharmaceutical dosage form was found very accurate with 99.37%.

 

DISCUSSION:

A modest and delicate spectroscopic process for quantitative estimation of Ondansetron in either pure form or in tablets unit dosage for was established. Ondansetron showed wavelength maxima at 302nm in ethanol. The developed method was showed the linear retort in the entire range of 4 to 24μg per ml with correlation coefficient of 0.9991. The lineddeterioration equation gottenwas Y = 0.037x + 0.022. The techniquewas shown the very good exactness the RSD was less than 2% and accuracy is 99.32±0.66. No substantialintrusion was detected in the absorbance of Ondansetron in the incidence of shared excipients. The system was statistically authenticatedbestowing to ICH. The technique was engaged for the quantitative determination of tablet dosage form. In assumption,the established spectroscopic methods are simple, specific, and reproducible, and can be used in routine analysis of Ondansetron in bulk.

 

ACKNOWLEDGEMENT:

The authors acknowledge the organization of Quantum University and SGRR University, for their unremitting support and inspiration. Thanks are also owed to the Quantum School of Pharmaceutical Sciences for equipment services as long as towards carrying out the work.

 

CONFLICTS OF INTEREST:

The authors have declared no conflicts of interest.

 

REFERENCES:

1.      The merck index An Encyclopedia of Chemicals, Drugs and Biological 2001; 13^th Edn, Merck andCompany, Inc.1224.

2.      Varvara A, Crina MM, Corina A, Popescu C. Ion Pair Reversed-Phase High Performance Liquid Chromatography of Ondansetron Hydrochloride using Sodium HeptaneSulphonate as a counter ion. Farmacia. 2009:57(4): 442-451.

3.      Kumar PR Murali K, Prakash PB et al. Derivative Spectrophotometric Estimation of Ondansetron and Paracetamol. E-Journal of Chemistry. 2006;3(3): 134-136.

4.      Shrish P, Patel LJ. Derivative Spectrophotometric Method for Simultaneous Determination of Ondansetron andRabeprazole in Combined Dosage Form. Asian Journal of Pharmaceutical and Clinical Research. 2012; 5(2): 37-41.

5.      Pillai S, SinghviI. Spectrophotometric Simultaneous Estimation of Ranitidine Hydrochloride and Ondansetron Hydrochloride from Tablet S Formulation. Indian journal of Pharmaceutical Sciences, 2007; 69(4): 601-604.

6.      Asad R, Abdul S I, Ujma R. Spectrophotometric Determination of Ondansetron Hydrochloride in Pharmaceutical Bulkand Dosage Forms. Journal of the Chinese Chemical Society.2007; 54(1): 223-227.

7.      Lahuerta Z. et al, Extractive Spectrophotometric Determination of Ondansetron by Ion-Pair Formation with Bromocresol Green. Taylor Francis online.1996; 29(5): 785-795.

8.      Lin Y, Stewart JT. Determination of Ondansetron and Diphenhydramine Mixture in 0.9% Sodium Chloride Injection. Journal of Liquid Chromatography Related Technologies. 1996; 19(5): 711-718.

9.      Ding P, Xu H, Wei G, Zheng J. microdialysis Sampling coupled to HPLC for Tran dermal Delivery Studyo of Ondansetron Hydrochloride in Rats. Biomedical Chromatography. 2000; 14(3): 141-143.

10.   Ravi S, yusrida d, Nurzalina K. Development and validation of a RP-HPLC method for the determination of Ondansetron. Application to pharmaceutical dosage.2009; 70(1-20): 75-81.

11.   Andreea V, Crina, Monciu et al. Application of a Selective Bonded Phase in the Liquid Chromatographic Assay of Ondansetron Hydrochloride and its impurities. Farmacia. 2009; 57(1): 1-5.

12.    Siluveru M,  Stewart J T. Enantiomer selective determination of S-(+) - and R-(-)-Ondansetron in Human Serum using Derivatized Cyclodextrins - Modified Capillary Eectrophoresis and Solid-Phase Extraction. Journal Chromatography- B.1997; 691(1):217-222.

13.   Shirish R P, Patel J, Yogeshvar P. Development and Validation of Analytical Method for the Determination of Rabeprazole and Ondansetron in Pharmaceutical Dosage Form by RP-HPLC. International Journal of Chemical Technology Research. 2010;2(3):1531-1536.

14.   Raval PB, Wadher SJ, Yeole PG. A Validated HPTLC Method for Determination of Ondansetron in Combination with Omeprazole or Rabeprazole in Solid Dosage Form. Indian journal of pharmaceutical sciences. 2008; 70(3): 386-390.

15.   Dotsikas Y, Kousoulos C, Tsatsou G. Development and Validation of a rapid 96-well Format Based Liquid–liquid Extraction and Liquid Chromatography-Tandem Mass Spectrometry Analysis Method for Ondansetron in Human Plasma. Journal of Chromatography-B.2006; 83(6): 79-82.

16.   Lingxia P, Qing W, Youpei W. Development and Validation of LC-MS/MS Method for Determination of Ondansetron in Rat Plasma and its Application. Latin American Journal of Pharmacy. 2012;31(2): 305-309.

17.   International Conference on Harmonization (ICH) Validation of Analytical Procedures: Text and methodology - Q2 (R1), 2005.

 

 

 

Received on 04.06.2020            Modified on 14.07.2020           

Accepted on 18.08.2020      ©Asian Pharma Press All Right Reserved