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Madhuri Sharma et al. / International Journal of Drug Research and Technology 2016, Vol. 6 (2), 34-42.
Accuracy was determined by replicate analysis of The standard deviation of the peak is calculated
samples containing known amount of 80 - 120 % for each parameter and the %RSD was found to be
of the target amount. The percentage recovery was less than 2% (Table-2).
98.45 – 101.0 % for CLP and 99.8 – 99.38 % for System Suitability
BNZ accordingly. Various System Suitability parameters were
Precision calculated. The parameters were found within
Precision describes the repeatability of analytical acceptance criteria (Table- 3).
method. Six aliquots of each single concentration Quantitative Determination in Gel Formulation
were prepared and analyzed on same day and In a quantitative determination % assay of CLP
three consecutive days. The developed method was found to be 99.9% and BNZ was found to be
was found to be precise as the %RSD were < 2 %. 99.7% (Table-4).
Precision of the method was determined by Summary of all the validation parameter has been
intraday and inter-day studies. The different mention hear (Table-5).
concentration of CLP 10.0 µg/ml - 30.0 µg/ml
their RSD is less than 2 that is between 0.10 – CONCLUSION
0.41 and BNZ different concentration is taken Development and validation of RP-HPLC method
25.0 µg/ml – 75.0 µg/ml their RSD is less than 2 was found to be simple, specific, robustness,
that is between 0.35 – 0.86, Whereas inter-day accurate, precise and economical. These methods
different concentration of CLP 10.0 µg/ml - 30.0 can be applied for routine quantitative analysis of
µg/ml their RSD is less than 2 that is between Clindamycin Phosphate and Benzoyl Peroxide in
0.61 – 0.84 and BNZ different concentration is gel formulation. This method was validated as per
taken 25.0 µg/ml – 75.0 µg/ml their RSD is less ICH guideline (Q2R1). For RP-HPLC linearity of
than 2 that is between 0.66-1.04 . Clindamycin Phosphate and BNZ were found in
the range 10.0-30.0 µg/ml and 25.0-75.1 µg/ml.
Robustness
The robustness of an analytical procedure is a ACKNOWLEDGEMENT
measure of its capacity to remain unaffected by We are thankful to Insta Pharmaceutical, Baroda
small but deliberate variations in the analytical and Valeant Pharmaceuticals, USA for providing
procedure parameters [pH (±0.2), Flow rate (±0.2 gift samples of Clindamycin Phosphate and
ml) and proportion of mobile phase (±2.0 v/v]. Benzoyl Peroxide for research work.
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Madhuri Sharma et al. / International Journal of Drug Research and Technology 2016, Vol. 6 (2), 34-42.
210 nm
210 nm 236 nm
Figure 5: Chromatogram of standard mixture for CLP and BNZ (20 μg/ml and 50 μg/ml)
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Madhuri Sharma et al. / International Journal of Drug Research and Technology 2016, Vol. 6 (2), 34-42.
Figure 8: Chromatogram of standard CLP (20µg/ml) and BNZ (50 µg/ml) using mobile phase 20mM
Ammonium acetate buffer pH 4.0: Methanol (45:55 %v/v).
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Madhuri Sharma et al. / International Journal of Drug Research and Technology 2016, Vol. 6 (2), 34-42.
Figure 9: Chromatogram of CLP (20µg/ml) and BNZ (50 µg/ml) of gel formulation using mobile phase 20
mM Ammonium acetate buffer pH 4.0: Methanol (45:55 %v/v)
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Madhuri Sharma et al. / International Journal of Drug Research and Technology 2016, Vol. 6 (2), 34-42.
Table 5: Regression analysis data & summary of validation parameter for proposed method
Parameters Results
CLP BNZ
Linear Range (n=3) (µg/ml) 10.0-30.0 25.0-75.1
Slope 197421 165222
Intercept 18169 28381
Limit of Detection(µg/ml) 0.32 0.72
Limit of Quantification (µg/ml) 0.98 2.19
Regression equation y = 197421x – 18169 y = 165222x – 28381
2
Correlation co-Efficient(r ) 1.000 1.000
Assay recovery (%) 98.45-101.0 99.8-99.38
Repeatability (%RSD) 0.4 0.3
Precision (n=3)
Intra-day (%RSD) 0.10-0.41 0.35-0.86
Inter-day (n=3) 0.61-0.84 0.66-1.04
Specificity Specific Specific
Robustness Robust Robust
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Correspondence Author:
Madhuri Sharma
Sat Kaival College of Pharmacy, Sarsa Crossroads, Sarsa-388365 Ta. Dist. Anand, Gujarat, India
Cite This Article: Madhuri, Sharma and Ankita, Bhavsar (2016), “Development and validation of HPLC
method for simultaneous estimation of clindamycin phosphate and benzoyl peroxide in gel formulation”,
International Journal of Drug Research and Technology, Vol. 6 (2), 34-42.
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