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Development and Validation of RP-HPLC Method for Simultaneous Estimation of Ibuprofen and Chlorzoxazone in Synthetic Mixture


ANALYSIS OF IBU AND CHL IN SYNTHETIC MIXTURE:-
The response of the sample solution was measured at 221 nm under the chromatographic condition mentioned above for the quantification of IBU and CHL. The amounts of IBU and CHL present in sample solution were determined by applying values of the peak area to the regression equations of the calibration curve.

RESULTS AND DISCUSSION:-

Linearity
Linear correlation was obtained between peak area Vs concentrations of IBU and CHL in the concentration of 2-30 µg/ml for IBU and 2-30 μg/ml for CHL. Regression parameters are mentioned in Table and the calibration curves of these two drugs at 221 nm are shown in Fig 5 and 6.

Method Precision (% Repeatability)
The RSD values for IBU and CHL were found to be 1.95 and 0.34 %, respectively . The RSD values were found to be <2 %, which indicates that the proposed method is repeatable.

Intermediate Precision (Reproducibility)
The low RSD values of interday and intraday for IBU and CHL, respectively, reveal that the proposed method is precise .

LOD and LOQ
LOD values for IBU and CHL were found to be  0.96  µg/mland 2.92 µg/ml, respectively and LOQ values for IBU and CHL were found to be 0.69µg /ml and 2.09µg/ml, respectively (Table 5). These data show that the proposed method is sensitive for the determination of IBU and CHL.

Accuracy
The recovery experiment was performed by the standard addition method. The recoveries obtained were 98.57 ± 0.49% and 99.09 ± 0.85 % for IBU and CHL, respectively (Table 3). The low value of standard deviation indicates that the proposed method is accurate. Results of recovery studies are shown in Table 3.

Assay of the synthetic mixture
The proposed validated method was successfully applied to determine IBU and CHL in their synthetic mixture. The result obtained for IBU and CHL was comparable with the corresponding labeled amounts (Figure 7) (Table 4)

Table 1 System Suitability Parameters of Chromatogram for IBU and CHL

Parameters

IBU ± RSD

(n = 6)

CHL ± RSD

(n = 6)

Retention time (min)

5.40±0.29

3.31± 0.20

Tailing factor

1.06± 1.24

1.10± 1.13

Theoretical plates

      3357± 1.65

      2664± 1.61

Resolution

4.58 ± 1.51

                             Table 2 Recovery Data for the proposed Method

Drug

Level

Amount of sample taken (µg/ml)

Amount of standard   spiked (%)

Mean% Recovery ± SD

IBU

I

8

50 %

 

98.05 ± 0.66

 

       II

8

100 %

 

98.65 ± 0.90

      III

8

150 %

99.03 ± 0.80

CHL

I

10

50 %

 

100.00 ± 1.87

 

II

10

100 %

 

98.93 ± 0.74

 

III

10

150 %

98.32 ± 0.99

Table 3 Analysis of IBU and CHL (n = 6)

Sample No.

Label Claim

Amount Found

% Label Claim

IBU

(mg)

CHL

(mg)

IBU

(mg)

CHL

(mg)

IBU

 (%)

CHL

 (%)

1

40

50

39.87

50.05

99.68

100.11

2

40

50

39.95

49.64

99.87

99.29

3

40

50

39.22

49.42

98.05

98.84

4

40

50

39.39

49.41

98.49

98.82

5

40

50

39.24

49.01

98.12

98.03

6

40

50

39.56

49.50

98.92

99.01

Mean

39.53

49.51

98.84

99.02

S.D.

0.311

0.34

0.77

0.68

Table 4  Regression Analysis Data and Summary of Validation Parameter for the proposed Method

Parameters

RP-HPLC method

IBU

CHL

Detection wavelength(nm)

221

221

Concentration range (µg/ml)

2-30

2-30

Regression equation

Y= mX + c

Y= 72937X + 35567

Y=66004X + 81349

Correlation coefficient

0.997

0.997

LOD(µg/ml)

0.96

0.69

LOQ(µg/ml)

2.92

2.09

% Recovery (Accuracy, n = 3)

98.57 ± 0.49

 99.09 ± 0.85

Repetability (% RSD, n = 6)

1.95

0.34

Precision (%RSD)

 

Interday (n = 3)

0.61-1.85

0.57-1.26

Intraday (n = 3)

0.11-1.12

0.24 - 0.96

 

CONCLUSION:-
In this proposed method the linearity is observed in the concentration range of 2 to 30 µg/ml and 2 to 30 µg/mlwith co-efficient of correlation, (r2) = 0.997 and (r2) = 0.997 for IBU and CHL, respectively at 221 nm. The result of the analysis of pharmaceutical formulation by the proposed method is highly reproducible and reliable and it is in good agreement with the label claim of the drug. The method can be used for the routine analysis of the IBU and CHL in synthetic mixture without any interference of excipients. 

ACKNOWLEDGEMENT
The authors are thankful to Acme Pharmaceutical Ltd., Ahmedabad, India for providing gift sample of IBU and CHL for carry out the research work.

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