1. INTRODUCTION:

YOSPRALA is newly designed tablet which effective in cardiovascular as well as gastrointestinal protection due to its immediate release of Omeprazole (40mg) and delayed release of Aspirin (81mg) or (325mg) dose strength. Yosprala is approved by USFDA in Sept 2016 for cardiovascular and cerebrovascular diseases.

On extensive literature survey it was found that very few methods are reported for Simultaneous estimation of Aspirin and Omeprazole in combined dosage form by any analytical technique. There are methods developed on single Aspirin only or combination with other drugs by using UV spectroscopy in tablet dosage form, by HPTLC method. Hence I was decided to develop a new method which having accurate, precise, economical, rapid and cost effective method for simultaneous estimation of Aspirin and Omeprazole in Tablet formulation and proposed method was validated according to ICH guidelines.

Figure.1 Aspirin

Aspirin, 2-(acetyloxy) benzoic acid, acts as an inhibitor of cyclooxygenase which results in the inhibition of the biosynthesis of prostaglandins. It also inhibits platelet aggregation and is used in the prevention of arterial and venous thrombosis.

Figure. 2 Omeprazole

Omeprazole is a proton pump inhibitor used in the treatment of dyspepsia, peptic ulcer disease (PUD), Gastro esophageal reflux disease (GORD/GERD), Laryngopharyngeal reflux (LPR) and Zollinger–Ellison syndrome.

Omeprazole is entirely metabolised by the hepatic cytochrome P-450 system (CYP), mainly in the liver. Identified metabolites in plasma are the sulfone, the sulfide and hydroxyomeprazole.

Simultaneous equation method is used where a sample contain two absorbing drugs (X and Y) each of this absorbance λ_maxof each other, it may be possible to determine both the drugs by the technique of simultaneous equation method.

2. MATERIALS AND METHODS

The bulk drug of pure Aspirin was provided by Alta laboratories, Mumbai and Omeprazole from Blue Cross Laboratories, Mumbai as gift sample and used as such. Fixed dose combination tablets (Yosprala) tablets containing ASP (325 mg) and OMZ (40 mg) was procured fromlocal Market. All chemicals and reagents of analytical grade and HPLC grade were purchase from Merck Chemical, Mumbai, INDIA

2.1 Instrument:

A double beam Shimadzu UV-visible spectrophotometer model 1800 with UV probe software was used for absorption measurements.

2.2 Solubility:

As per solubility studies it was found that both the drug sample are freely soluble in methanol.

2.3Determination of λ Max of Individual Component:

An appropriate aliquot portion of ASP (0.5 mL) and OMZ (0.1 mL) were transferred to two separate 10 mL volumetric flasks, the volume was made up to the mark using 80% v/v methanol to obtain ASP (50 µg/mL) and OMZ (10 µg/mL). Drug solutions were scanned separately between 200 nm to 400 nm.

The overlain spectrum of both drugs having concentrations 130 µg/mL ASP and 16 µg/mL OMZ was recorded and two wavelengths 276.0 nm (λ max of ASP) and 301.0 nm (λ max of OMZ) were selected for further study.

2.4Preparation of Standard Stock Solutions of ASP and OMZ

A) Aspirin Standard Stock Solution [A]:

An accurately weighed quantity of ASP (10 mg) was taken in 10 mL volumetric flask and dissolved in methanol (8 mL) with the help of ultrasonication for about 10 min. Then the volume was made up to the mark using methanol to get Aspirin standard stock solution (1 mg / mL).

B) Omeprazole Standard Stock Solution [O]:

An accurately weighed quantity of OMZ (10 mg) was taken in 10 mL volumetric flask and dissolved in methanol (8 mL) with the help of ultrasonication for about 10 min. Then the volume was made up to the mark using methanol to get Omeprazole standard stock solution (1 mg / mL).

Table 1: Linearity Study of ASP at 276 nm.

Sr. No.	Concentration of ASP in [µg/mL]	Absorbance Mean ± S.D. [n = 5]	% R.S.D.
1	10	0.073±0.0008	1.142
2	20	0.112±0.0007	0.631
3	30	0.165± 0.0018	1.133
4	40	0.212±0.0011	0.536
5	50	0.265±0.0008	0.315

Table 2: Linearity Study of OMZ at 301 nm.

Sr. No.	Concentration of OMZ in[µg/mL]	Absorbance Mean ± S.D. [n = 5]	% R.S.D.
1	2	0.0588 ± 0.0018	1.117
2	4	0.1052 ± 0.0019	1.828
3	6	0.1622 ± 0.0014	0.914
4	8	0.225 ± 0.0012	0.544
5	10	0.2744 ± 0.0018	0.662

Figure 3: Calibration Curve of Aspirin at 276 nm

Figure 4: Calibration Curve of Omeprazole

Figure 5: Overlay Spectra of ASP at 276 nm

Figure 6: Overlay Spectra of OMZ at 301nm

3. RESULT AND DISCUSSION

Results of Standard Laboratory Mixture (API) and YOSPRALA Tablet by UV- Spectroscopic methods

Table 3: Summary of Results for UV-spectroscopic methods

Sr. No	Sample	Statistical data	%Label Claim
Sr. No	Sample	Statistical data	ASP	OMZ
1.	Standard Laboratory Mixture (API)	Mean	99.85	99.80
		S.D.	0.0764	0.0697
		% R.S.D	0.0766	0.0699
2.	YOSPRALA Tablet	Mean	99.87	99.85
		S.D.	0.0702	0.0465
		% R.S.D	0.0703	0.0466

4. VALIDATION OF PROPOSED METHOD:

The Proposed method was validated as per the ICH guidelines.

4.1 Accuracy [Recovery Study]:

Accuracy of proposed method was ascertained on the basis of recovery study performed by standard addition method

Table 4: Recovery Study

Tablet- YOSPRALA Average Weight of Tablet-478 mg.
Sr. No.	Quantity Tablet Powder Taken	Percentage %	Amount of Pure Drug Added (mg)		Total Amount of Drug Recovered (mg) ± S.D. (n = 3)		% of Drug Recovered (n = 3)
Sr. No.	Quantity Tablet Powder Taken	Percentage %	ASP	OMZ	ASP	OMZ	ASP	OMZ
1.	478	80	260	32	583.3 ± 1.08	71.64± 0.06	99.70	99.50
2.	478	100	325	40	649.6 ± 1.17	79.91 ±0.05	99.95	99.88
3.	478	120	390	48	714.7 ± 1.43	87.95 ± 0.02	99.90	99.94
						Mean	99.85	99.77
						SD	0.1322	0.2386
						% RSD	0.1324	0.2388

4.2 Precision:

Table 5: Precision Study

Drug	Conc. [µg/mL]	Intra-day (Amount Found)		Inter-day (Amount Found)
Drug	Conc. [µg/mL]	Mean ±S.D [n = 5]	% R.S.D.	Mean ± S.D. [n = 5]	% R.S.D.
ASP	10	9.94 ± 0.064	0.6462	9.92 ± 0.1013	1.0213
	20	19.89 ± 0.1814	0.9121	19.86 ± 0.2309	1.1622
	30	29.79 ± 0.272	0.9156	29.75 ± 0.3614	1.2147
OMZ	2	1.97 ± 0.081	0.5452	1.95 ± 0.0151	0.7753
	4	3.92 ± 0.0336	0.856	3.91 ± 0.0420	1.0737
	6	5.87 ± 0.0342	0.5819	5.83 ± 0.0707	1.2128

According to ICH guidelines acceptance criteria for precision the %RSD should NMT 2%

4.3 Ruggedness:

Table 6: Ruggedness Study

Parameters	ASP 325 µg/mL		OMZ 40 µg/mL
Parameters	Amount Found in µg/mL Mean ± S.D. (n = 3)	% RSD	Amount Found in µg/mL Mean ± S.D.(n = 3)	% RSD
Analyst I	324.46± 0.3614	0.1113	39.78 ± 0.1484	0.3730
Analyst II	324.95 ± 0.3968	0.1221	39.82 ± 0.1569	0.3940
Day-I	324.29 ± 0.6340	0.1955	39.77 ± 0.1861	0.4678
Day-II	325.06 ± 0.6562	0.2019	39.80 ± 0.1908	0.4795
Instrument I	324.21 ± 0.7379	0.2276	39.73 ± 0.2753	0.6930
Instrument II	324.89 ± 0.8580	0.2641	39.70 ± 0.2662	0.6707

4.4 LOD:

Limit of detection of Aspirin and Omeprazole were found to be 0.2528 μg/mL and 0.2307 μg /mL respectively.

4.5LOQ:

Limit of Quantitation of Aspirin and Omeprazole were found to be 1.766 μg/ mL and 1.954 μg/mL respectively.

4.6 System suitability parameters:

The following parameters are system suitability parameters for the analytical method developed according to ICH guidelines.

Table 7: System suitability parameters

Sr.no	Parameters	Aspirin	Omeprazole
1	λmax	276nm	301nm
2	Regression co-efficient (r2)	0.997	0.997
3	LOD (μg/ml)	0.2528	0.2307
4	LOQ (μg/ml)	1.766	1.954
5	Linearity range	10-50μg/ml	2-10μg/ml

5. CONCLUSION:

It was made to develop an analytical method for simultaneous Equation method for estimation of Aspirin and Omeprazole by using UV Spectroscopy. The developed method was validated for linearity, Accuracy, precision, ruggedness and results were within the limits according to ICH guidelines. The proposed method was cost effective, simple, rapid, economic, cheap, Precise and robust. The above method can be used for routine analysis of Aspirin and Omeprazole in bulk and Tablet Dosage Form

6. ACKNOWLEDGEMENT:

The author’s were thankful to Principle of TVES’s HLMC College of Pharmacy, Faizpur (MS) INDIA for providing necessary help for work.

7. REFERENCES:

1. Yosprala prescribing information. Princeton, NJ: Aralez pharmaceuticals US Inc.; 2016.

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