DEVELOPMENT AND VALIDATION OF A UV SPECTROPHOTOMETRIC METHOD FOR SIMULTANEOUS ESTIMATION OF NAPROXEN AND SUMATRIPTAN

www.wjpr.net Vol 7, Issue 01, 2018. 1418

DEVELOPMENT AND VALIDATION OF A UV

SPECTROPHOTOMETRIC METHOD FOR SIMULTANEOUS

ESTIMATION OF NAPROXEN AND SUMATRIPTAN

1

*Vernekar Saumya and 2Dr. Theivendren Panneerselvam

1*

Asst Professor, Dept of Pharmaceutical Chemistry, PES’s Rajaram and Tarabai Bandekar,

College of Pharmacy, Ponda Goa India.

2

Professor, Karavali College of Pharmacy, Mangalore Karnataka India.

ABSTRACT

Naproxen is a non-steroidal anti-inflammatory drug (NSAID)

commonly used for reduction of moderate to severe pain, fever,

inflammation and stiffness. Sumatriptan is a 5HT agonist. Acting as an

agonist at these receptors, Sumatriptan reduces the vascular

inflammation associated with migraines. The objective of the present

work was to develop and validate a simple UV method for

simultaneous estimation of Naproxen and Sumatriptan in combined

dosage form.Q Absorption ratio method was used for calculation of

assay results. Methanol and Phosphate buffer pH 6.8 (50:50 v/v) was

used as a diluent. The developed method was validated according to

ICH guidelines and found to be specific, accurate, precise and robust.

Assay of the formulation was performed and the mean assay values were found to be within

acceptable limits. These methods can also be used to estimate NAP and SUM as independent

components. The developed methods are cost effective, time saving, simple and precise.

KEYWORDS: Naproxen, Sumatriptan, UV method, Q Absorption Ratio method.

1. INTRODUCTION

Naproxen is used for reduction of pain, fever, inflammation and stiffness caused by migraine,

osteoarthritis, kidney stones, gout, and menstrual cramps. It works by inhibiting COX-1 and

COX-2 enzymes.[1, 2] They work by reducing the levels of prostaglandins, chemicals that are

responsible for pain, fever, and inflammation.[3,4,5] Sumatriptan is a synthetic drug belonging

Volume 7, Issue 01, 1418-1427. Research Article ISSN 2277–7105

Article Received on 22 Nov. 2017,

Revised on 12 Dec. 2017, Accepted on 01 Jan. 2018

DOI: 10.20959/wjpr20181-10586

*Corresponding Author Vernekar Saumya

Asst Professor, Dept of

Pharmaceutical Chemistry, PES’s Rajaram and Tarabai

Bandekar, College of

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to the triptan class, used for the treatment of migraine headaches.[6,7,8] It is structurally similar

to serotonin (5HT), and is a 5-HT agonist.[9,10] Sumatriptan reduces the vascular inflammation

associated with migraines. Hardly few techniques are available for this combination which

could be used for determination over a wide linear range.[11] Main objective is to develop and

validate[12] UV – Visible method using Absorption Q Ratio method which is accurate and

economical.

O

ONa

O

Fig 1: Structure of Naproxen sodium.

N H

N

S N

H O

O

COOH

COOH

.

Fig 2: Structure of Sumatriptan succinate.

2. MATERIALS AND METHOD

UV/Visible spectrophotometer (Shimadzu Model UV 1800) was employed with spectral

bandwidth of 1nm and wavelength accuracy of 0.3 nm (with automatic wavelength correction

with a pair of 1 cm matched quartz cells). Mettler M80 analytical balance was used for

weighing of active and placebo. Analytical grade reagents and solvents were used for the

study; Naproxen and Sumatriptan were procured as free gift samples. Tablet dosage form

HEADSET was bought from local market 100% methanol (Rankem, HPLC grade) was used

as diluent 1. Buffer pH 6.8 was prepared by taking 6.8g of potassium dihydrogen phosphate

in 1000ml of milliQwater. pH adjusted to 6.8 with dilute sodium hydroxide solution. 50:50%

v/v of buffer pH 6.8 and methanol was used as diluent 2. This was mixed in equal ratio and

www.wjpr.net Vol 7, Issue 01, 2018. 1420 2.1. Preparation of standard solution

39.34mg of Naproxen and 7.15mg of Sumatriptan was weighed accurately and transferred

into a dry 100ml volumetric flask. To this 70ml of diluent 1 was added and it was sonicated

for about 10mins. Volume was made up to mark with diluent 1. This was centrifuged for

10mins at 5000rpm. 6.25ml of supernatant was transferred to a 25ml volumetric flask.

Volume was made up to mark with diluent 2 and mixed thoroughly. The final concentration is 98.36 and 17.86 μg/ml of Naproxen and Sumatriptan resp.

2.2. Preparation of placebo solution

994.962mg of placebo equivalent to one tablet was weighed and transferred to 100ml

volumetric flask. This was treated as same as that of standard solution.

2.3. Preparation of test sample

10 tablets are weighed and average weight was taken. These tablets were finely crushed in

mortar and pestle and this powder was used for analysis. Weigh tablet powder containing

40mg and 6.8mg of Naproxen and Sumatriptan equivalent to 400ppm and 68ppm above

drugs respectively and transfer to a 100ml volumetric flask. To this 70ml of diluent 1 was

added and it was sonicated for about 10mins. Volume was made up to mark with diluent 1

and this solution was centrifuged for about 10mins at 5000 rpm. 6.25ml of supernatant was

transferred in a 25ml flask and volume was made up with diluent 2. This was mixed well and

used for assay of the formulation.

3. METHOD VALIDATION

Method was validated for Linearity, Precision, Accuracy, Robustness, LOD and LOQ.

3.1. Instrument parameters: UV-Vis spectrophotometer

Measurement mode: Photometric.

Wavelengths selected: 262nm (λmax of Naproxen) and 277nm (iso-absorptive point).

Scan speed: Medium.

Cuvettes: quartz cell with 1 cm pathlength.

Software used: UV probe.

3.2.Method for analysis

Blank solution is put in both the cuvettes, photometric mode is selected in the method file and

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nm. Then autozero is selected and instrument ready for analysis. Read blank for correction.

Now put the standard solution in sample compartment and take absorbance’s at the required

wavelengths. Take 6 continuous readings to check for system suitability and RSD should not

be more than 2%. Same is done for placebo and test sample for assay. Each reading is taken

in triplicate.

Fig 3: Overlain spectra of Naproxen and Sumatriptan.

4. RESULTS AND DISCUSSION

4.1.Assay

The % Assay found is between 97% to 103% for both Naproxen and Sumatriptan in bulk as

well as in dosage form. It passes the assay limit as per the compendial specifications.

If the main components are present in salt form then weight equivalent to pure drug should be

taken.

Table 1: Assay of Naproxen and Sumatriptan by proposed UV method.

Sr.no

Amount of drug present (μg/ml)

Amount of drug found

(μg/ml) % Assay

NAP SUM NAP SUM NAP SUM

Test 1 100.53 17.10 98.75 16.89 98.23 98.77

Test 2 100.45 17.08 98.64 16.93 98.20 99.12

Test 3 100.63 17.10 98.69 16.83 98.10 98.42

4.2. Specificity

Placebo doesn’t interfere at the absorbing wavelengths. Placebo equivalent to 1 tablet is taken

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Also force degradation studies are done by treating in stress conditions and observing the

effect. The drugs are not affected by acid, alkali and water. However it undergoes excessive

oxidation with hydrogen peroxide due to presence of many oxidizable groups in both the

drugs.

Fig 4: Placebo interference for Specificity studies.

Fig 5: Forced degradation studies with hydrogen peroxide.

4.3. Linearity

Naproxen is found linear in range of 10 to 110 μg/ml. Sumatriptan is found linear in range of 10 to 110 μg/ml. Correlation coefficient is more then 0.999. The linearity was done for

www.wjpr.net Vol 7, Issue 01, 2018. 1423 Fig 6: Linearity curve of Naproxen at 277nm.

Fig 7: Linearity curve of Naproxen at 262nm.

Fig 8: Linearity curve of Sumatriptan at 277nm.

277nm

262nm

www.wjpr.net Vol 7, Issue 01, 2018. 1424 Fig 9: Linearity curve of Sumatriptan at 262nm.

4.4. Accuracy

Accuracy studies were done by doing recovery studies under presence of placebo. Recovery

was done at low, medium and high level ie. 50%, 100% and 150%. Recoveries are found in

the limit and thus the range is from 97% to 99% for Naproxen. For Sumatriptan it’s from

98% to 100%.

Table 2: Data for Accuracy studies by Recovery.

Ac

cu

rac

y

leve

l (%)

Amount of API added (μg/ml)

Amount of API

recovered (μg/ml) % Recovery % RSD

NAP SUM NAP SUM NAP SUM NAP SUM

50 50.06 8.31 48.83 8.22 97.8 98.9 0.46 1.4 100 99.05 17.13 97.6 16.45 98.5 97.3 0.20 1.5 150 150.60 25.4 146.48 24.68 97.8 97.2 0.27 1.4

4.5. Precision

It is carried out by doing repeatability study which showed RSD below 2% indicating that the

method is precise. Also intermediate precision is done and found suitable.

Solution stability is carried out to see the effect of storage on the sample. Solution is found

stable upto 48 hours and after that the degradation is NMT 6% as that compared to control.

www.wjpr.net Vol 7, Issue 01, 2018. 1425 Table 3: Data for Precision studies.

Precision studies Acceptance criteria Observation

Solution stablility %RSD NMT 2% Stable upto 48hrs at room temperature

Repeatability Individual assay 97-103% %RSD NMT 2%

Average Assay results 97-99% %RSD 0.2%

Intermediate precision

Intra Day study Individual assay 97-103% %RSD NMT 2%

Average Assay results 97-100% %RSD 0.8%

Inter Analyst study Individual assay 97-103% %RSD NMT 2%

Average Assay results 97-100% %RSD 0.1%

4.6. LOD (Limit of Detection)

It’s done according to the readings taken from calibration curve and it’s a not that much

required criteria for assay method. Values found from calibration curve are as follows.

Naproxen (262nm): 3.01, Naproxen (277nm): 1.83, Sumatriptan (262nm): 3.84, Sumatriptan

(277nm): 3.41.

4.7.LOQ (Limit of Quantitation)

Values found from calibration curve are as follows.

Naproxen (262nm): 9.14, Naproxen (277nm): 5.56, Sumatriptan (262nm): 11.65, Sumatriptan

(277nm): 10.33.

4.8. Robustness

To check for any variability allowed in the method robustness is a very important criterion to

be considered. For UV method wavelength change is done and also diluents change. The

results do not affect the method and it is found to be robust.

Allowable wavelength variation is from -1 to +1 nm. Allowable variation in diluent

composition is 10% of the organic phase. The %RSD for both the variations is below 2%.

5. CONCLUSIONS

A novel and simple Q absorption ratio method has been developed for simultaneous analysis

of Naproxen and Sumatriptan in tablet formulation. Simple, Accurate, Precise, Economical

UV method has been developed using phosphate buffer pH 6.8 and methanol (50:50 v/v) as

diluent. Determination is done at 262.0 and 277.0 nm over the concentration ranges of 10-110

µg/ml for both. Validation also covers parameters like Accuracy, Specificity, Robustness,

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