ABSORBANCE CORRECTION METHOD FOR SIMULTANEOUS ESTIMATION OF AMLODIPINE BESYLATE AND SIMVASTATIN IN SYNTHETIC MIXTURE

Pharma Admission

pharma admission

 

Table.2 Intraday precision data for estimation of AML,and SIM *(n=3)

Precision

Con

AML

Con

SIM

Abs. ±% RSD

5 µg/ml

0.093 ± 0.22

5 µg/ml

0.619± 0.16

15 µg/ml

0.275± 0.41

15 µg/ml

1.847± 0.11

25 µg/ml

0.449± 0.12

25 µg/ml

2.813± 0.12

ii. Interday Precision
P
rocedure
- Theprecisionofthedevelopedmethodwasassessedbyanalyzing combined standard solution  containing three different concentrations 5,15,25  μg/ml  for AML and 5,15,25 μg/ml  for SIM  triplicate (n=3) per day for consecutive 3 days for inter-day precision.
- For zero order spectraabsorbancewas measured at 360.80 nm for AMLand 237.60  nm forSIM. The  %  RSD  value  of  the  results  corresponding  to  the  absorbance  was expressed forinter-dayprecision.

Table.3 Interday precision data for estimation of AML,and SIM*(n=3)

Precision

Con

AML

Con

SIM

Abs. ±% RSD

5 µg/ml

0.094± 0.32

5 µg/ml

0.623± 0.27

15 µg/ml

0.278 ± 0.54

15 µg/ml

1.850± 0.13

25 µg/ml

0.451± 0.22

25 µg/ml

2.816± 0.16

3) ACCURACY
• Accuracy of the method was determined by recovery study from synthetic mixture at three levels (80%, 100%, and 120%) of standard addition.
• The % recovery values are tabulated in Table and Percentage recovery for AMLO and SIM by this method was found in the range of 100.37 to 102.12% and 98-101.81% respectively,
• The value of %RSD within the limit indicated that the method is accurate and percentage recovery shows that there is no interference from the excipients.

Table.4 Recovery data of AML*and SIM*(n=3)

Initial conc. (µg/ml)

Level

of recovery

Quantity of Std. Added

(µg/ml)

Total       Amount

(µg/ml)

Result of recovery study

Total Quantity Found*

(µg/ml)± %RSD

% recovery ± %RSD

AML

SIM

AML

SIM

AML

SIM

AML

SIM

AML

SIM

10

10

0%

-

-

10

10

10.01±0.15

 10.12±0.11

100.41
±0.51

100.55
±0.49

10

10

80 %

8

8

18

18

18.02±11

17.86±14

100.62
±0.45

99.50
±0.47

10

10

100 %

10

10

20

20

20.14±15

20.09±21

100.80
±0.16

100.65
±0.29

10

10

120 %

12

12

22

22

22.05±22

22.08±18

100.46
±0.41

100.49
±0.44

Mean of 3 Determination

100.62%

100.21%

4) LOD & LOQ
· The Limit of detection and quantitation of the developed method was assessed by analyzing 10 replicates of standard solutions containing concentrations 5 μg/ml for AML and SIM.
· The LOD and LOQ were calculated as LOD = 3.3*σ/S, and LOQ = 10*σ/S, where σ is the standard deviation of the lowest standard concentration and S is the slope of the standard curve.% RSD was calculated.

Table.5 LOD and LOQ data of AML and SIM*(n=10)

Drugs

LOD (µg/ml)

LOQ (µg/ml)

Amlodipine besylate

0.17

0.54

Simvastatin

0.10

0.32

APPLICATION OFTHEPROPOSED METHOD FOR ANALYSISOF EDAAND ARG IN SYNTHETIC MIXTURE (ASSAY)
- A zero order derivative spectrum of the resulting solution was recorded and absorbances at 360.80nm and  237.60nm were noted for estimation of AML and SIM, respectively.
- The concentration of AML and SIM in mixture was determined using the corresponding calibration graph. The results from the analysis of synthetic mixture containing Amlodipine besylate (10mg) and Simvastatin(10mg) in combination are presented.The percent assay shows that there is no interference from excipients and the proposed method can successfully applied to analysis of commercial formulation containing AML and SIM. The % assay values are tabulated.

Table. 6 Analysisdata ofcommercialformulation*(n=3)

             Drugs

% Assay ± SD

% RSD(n=3)

Amlodipine besylate

100.45 ± 0.0058

0.58

        Simvastatin

100.85 ± 0.0061

0.60

6) Robustness
- Robustness and Ruggedness of the method was determined by subjecting the method to slight change in the method condition, individually, the:

- Change in Wavelength (± 0.2 nm)
AML at 360.60 nm and 361 nm and SIM at 237.40 nm and 237.80 nm
- Change in instrument (UV-Vis Spectrophotometer model 1800 and 2450),
- Three replicates were made for the concentration (5,15,25μg/ml of AML and 5,15,25 μg/ml of SIM) with different stock solution preparation.
- % RSD was calculated.

Table 7: Robustness and Ruggedness data of  AML and SIM*(n=3)

Condition

Conc.

(µg/ml)

    Different    Instrument

Wavelength (±0.2 nm)

UV-2450

UV-1800

360.60 nm

361 nm

AML

Mean (n=3)

± % RSD

5

0.094±0.10

0.096±0.13

0.094±0.17

0.098±0.21

15

0.276±0.17

0.274±0.15

0.275±0.19

0.278±0.16

25

0.448±0.18

0.445±0.27

0.449±0.10

0.452±0.18



  SIM

Mean(n=3)

± %RSD


UV-2450

UV-1800

237.40 nm

237.80 nm

5

0.620±0.33

0.624±0.27

0.625±0.19

0.627±0.29

15

1.848±0.22

1.845±0.23

1.846±0.21

1.850±0.20

25

2.812±0.20

2.814±0.32

2.817±0.17

2.820±0.25

SUMMARY OF VALIDATION PARAMETER.

PARAMETERS

Absorbance correction method

Amlodipine besylate

Simvastatin

Concentration range(µg/ml)

5-25

5-25

Regression equation

y = 0.0157+0.015


y = 0.0756x - 0.0235


Correlation Coefficient(r2)

0.9991


0.9993


Accuracy(%Recovery) (n=3)

100.62%

100.21%

Intra-dayPrecision (%RSD)(n=3)

0.12-0.41

0.11-0.16

Inter-dayprecision (%RSD)(n=3)

0.22-0.54

0.16-0.27

LOD(µg/ml)

0.17

0.10

LOQ(µg/ml)

0.54

0.32

Ruggedness and Robustness

0.10-0.27

0.19-0.33

%Assay

100.45%

100.85%

CONCLUSION:
All the parameters are validated as per ICH guidelines for the method validation and found to be suitable for routine quantitative analysis in pharmaceutical dosage forms. The result of linearity, accuracy, precision proved to be within limits with lower limits of detection and quantification. Ruggedness and Robustness of method was confirmed as no significant were observed on analysis by subjecting the method  to slight change in the method condition. Assay results obtained by proposed method are in fair agreement.

ACKNOWLEDGEMENT:
We are sincerely thankful to Shree Dhanvantary Pharmacy College, Kim, Surat, for providing us Infrastructure facilities and moral support to carry out this research work. We are also thankful to SDPARC for giving us their special time and guidance for this research work. We also thank our colleagues for their helping hand.


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