validation

1. VALIDATION OF ASSAY
METHOD OF IBUPROFEN BY
UV SPECTROSCOPY
PRESENTED BY :
TANMOY DAS
B.PHARM
SEM-7
NETAJI SUBHAS CHANDRA BOSE INSTITUTE
OF PHARMACY
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2. CONTENT
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3. VALIDATION
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4. ASSAY
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5. IBUPROFEN
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C COOH
CH3
H
CHCH2
H3C
H3C
Molecular formula1 C13H18O2.
Molecular Weight 206.28 g/mol
Melting Range 75 to 780C
Solubility Freely soluble in acetone, chloroform, ethanol (95%) and ether, practically insoluble in water, it
dissolves in dilute solution of alkali hydroxide, carbonates
Apparent pKa 5.2

6. PHOSPHATE BUFFER
SOLUTION(PBS),PH-7.2
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0
0.1
0.2
0.3
0.4
0.5
0.6
240 260 280 300 320 340 360 380 400
UVAbsorbance
Wavelength (nm)
UV spectrum of Ibuprofen in PBS pH 7.2 ,concentration 20.0 micg/ml
Drug

7. SPECIFICITY
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0
0.1
0.2
0.3
0.4
0.5
0.6
240 260 280 300 320 340 360 380 400
UVAbsorbance
Wavelength (nm)
UV spectrum of pure Ibuprofen and Ibuprofen matrix
Drug Drug+Matrix

8. LINEARITY
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1
2
5
10
15
20
25
y = 28.87x - 0.1486
R² = 0.9997
0
5
10
15
20
25
30
0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9 1
Conc(mg/ml)
Abs (221nm)
Standard straight line of Ibuprofen in PBS pH 7.2 at
264nm
Conc (micg/ml) Abs (264nm)
1 0.0346
2 0.0791
5 0.1828
10 0.3467
15 0.5285
20 0.6913
25 0.8748
Slope = 28.8700
Intercept = – 0.1486

9. RANGE
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00.51
2
5
10
15
20
25
30y = 27.39x + 0.2035
R² = 0.9967
0
5
10
15
20
25
30
35
0 0.2 0.4 0.6 0.8 1 1.2
Conc(micg/ml)
Abs (221nm)
Standard straight line of Ibuprofen in PBS pH 7.2 at 264nm
for testing the range
Conc (micg/ml) Abs (264nm)
0.5 0.0123
1 0.0346
2 0.0791
5 0.1828
10 0.3467
15 0.5285
20 0.6913
25 0.8748
30 1.1369

10. PREECISION
Conc. of pure drug added
(mg/ml)
Conc. of drug obtained by
assay
(mg/ml)
Mean SD
RSD
(%)
SEM
1 0.0352
0.0342 0.00140 4.09 0.00081 0.0348
1 0.0326
10 0.3482
0.3532 0.00569 1.61 0.003310 0.3521
10 0.3594
25 0.8762
0.8738 0.00325 0.37 0.001925 0.8751
25 0.8701
10/13𝑅𝑆𝐷 =
𝑆𝐷
𝑀𝑒𝑎𝑛
𝑥100% and 𝑆𝐸𝑀 =
𝑆𝐷
𝑛
where n = 3.

11. ACCURACY
Solution Vol. of Soln. A
(mL)
Vol. of Soln. B
(mL)
Distilled Water
(mL)
Final conc.
Approx. (g/ml)
Conc. After assay
(g/ml)
S1 0.1 0.1 9.8 1 C1
S2 1.0 1.0 8.0 10 C2
S3 2.5 2.5 5.0 25 C3
S4 0.1 0.0 9.9 0.5 C4
S5 1.0 0.0 9.0 5 C5
S6 2.5 0.0 7.5 12.5 C6
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𝑅𝑒𝑐𝑜𝑣𝑒𝑟𝑦 𝑓𝑜𝑟 𝑆1 & 𝑆4 =
𝐶1 − 𝐶4
0.5
𝑥100%
𝑅𝑒𝑐𝑜𝑣𝑒𝑟𝑦 𝑓𝑜𝑟 𝑆2 & 𝑆5 =
𝐶2 − 𝐶5
5.0
𝑥100%
𝑅𝑒𝑐𝑜𝑣𝑒𝑟𝑦 𝑓𝑜𝑟 𝑆3 & 𝑆6 =
𝐶3 − 𝐶6
12.5
𝑥100%

12. Conc. of pure drug added
(mg/ml)
Conc. of drug obtained by assay
(mg/ml)
% Recovery Mean SD
RSD
(%)
0.5 0.508
101.60
101.53 0.90 0.890.5 0.512
102.40
0.5 0.503
100.60
5 5.039
100.78
100.32 0.52 0.525 4.988
99.76
5 5.021
100.42
12.5 12.511
100.09
99.99 0.09 0.0912.5 12.496
99.97
12.5 12.489
99.91
Overall 100.61 0.88 0.87
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Determination of Accuracy of the assay method
The overall % recovery was found to be 100.61 and the RSD of 0.87%. Therefore, the proposed assay
method has good accuracy within the range of 1 to 25 gml.

13. CONCLUSION
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