Development and Optimization of HPLC Analysis of Metronidazole, Diloxanide, Spiramycin and Cliquinol in Pharmaceutical Dosage Forms Using Experimental Design.
A new simple, sensitive, rapid and accurate gradient reversed-phase high-performance liquid chromatography with photodiode array detector (RP-HPLC-DAD) was developed and validated for simultaneous analysis of Metronidazole (MNZ), Spiramycin (SPY), Diloxanidefuroate (DIX) and Cliquinol (CLQ) using statistical experimental design. Initially, a resolution V fractional factorial design was used in order to screen five independent factors: the column temperature (°C), pH, phosphate buffer concentration (mM), flow rate (ml/min) and the initial fraction of mobile phase B (%). pH, flow rate and initial fraction of mobile phase B were identified as significant, using analysis of variance. The optimum conditions of separation determined with the aid of central composite design were: (1) initial mobile phase concentration: phosphate buffer/methanol (50/50, v/v), (2) phosphate buffer concentration (50 mM), (3) pH (4.72), (4) column temperature 30°C and (5) mobile phase flow rate (0.8 ml min-1). Excellent linearity was observed for all of the standard calibration curves, and the correlation coefficients were above 0.9999. Limits of detection for all of the analyzed compounds ranged between 0.02 and 0.11 μg ml-1; limits of quantitation ranged between 0.06 and 0.33 μg ml-1The proposed method showed good prediction ability. The optimized method was validated according to ICH guidelines. Three commercially available tablets were analyzed showing good % recovery and %RSD.
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Sinai University, El Arish, North Sinai, Egypt.|Department of Medicinal Chemistry, Faculty of Pharmacy, University of Tabuk, Tabuk, Saudia Arabia.
Mahmoud M
MM
Elkhoudary
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt dr_rasalam@yahoo.com.
Randa A
RA
Abdel Salam
Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
Ghada M
GM
Hadad
D000890:Anti-Infective Agents / Q000032:analysis* / Q000737:chemistry / Q000592:standards; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004304:Dosage Forms; D005663:Furans / Q000032:analysis; D008795:Metronidazole / Q000032:analysis; D015203:Reproducibility of Results; D012107:Research Design; D015572:Spiramycin / Q000032:analysis
D000890:Anti-Infective Agents; D004304:Dosage Forms; D005663:Furans; D008795:Metronidazole; D015572:Spiramycin; C004932:diloxanide furoate