title: Validated spectrophotometric and chromatographic methods for simultaneous determination of ketorolac tromethamine and phenylephrine hydrochloride.
abstract: This work describes five simple and reliable spectrophotometric and chromatographic methods for analysis of the binary mixture of ketorolac tromethamine (KTR) and phenylephrine hydrochloride (PHE). Method I is based on the use of conventional Amax and derivative spectrophotometry with the zero-crossing technique where KTR was determined using its Amax and (1)D amplitudes at 323 and 341nm respectively, while PHE was determined by measuring the (1)D amplitudes at 248.5nm. Method II involves the application of the ratio spectra derivative spectrophotometry. For KTR, 12μg/mL PHE was used as a divisor and the (1)DD amplitudes at 265nm were plotted against KTR concentrations; while - by using 4μg/mL KTR as divisor - the (1)DD amplitudes at 243.5nm were found proportional to PHE concentrations. Method III depends on ratio-difference measurement where the peak to trough amplitudes between 260 and 284nm were measured and correlated to KTR concentration. Similarly, the peak to trough amplitudes between 235 and 260nm in the PHE ratio spectra were recorded. For method IV, the two compounds were separated using Merck HPTLC sheets of silica gel 60 F254 and a mobile phase composed of chloroform/methanol/ammonia (70:30:2, by volume) followed by densitometric measurement of KTR and PHE spots at 320 and 278nm respectively. Method V depends on HPLC-DAD. Effective chromatographic separation was achieved using Zorbax eclipse plus C8 column (4.6×250mm, 5μm) with a mobile phase consisting of 0.05M o-phosphoric acid and acetonitrile (50:50, by volume) at a flow rate 1mL/min and detection at 313 and 274nm for KTR and PHE respectively. Analytical performance of the developed methods was statistically validated according to the ICH guidelines with respect to linearity, ranges, precision, accuracy, detection and quantification limits. The validated spectrophotometric and chromatographic methods were successfully applied to the simultaneous analysis of KTR and PHE in synthetic mixtures of different proportions and laboratory-made ophthalmic solution.
authors.affiliation: University of Alexandria, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department, Elmessalah, Alexandria 21521, Egypt. Electronic address: tbelaleg@yahoo.com.
authors.forename: T S
authors.initials: TS
authors.lastname: Belal
authors.affiliation: University of Alexandria, Faculty of Pharmacy, Pharmaceutical Chemistry Department, Elmessalah, Alexandria 21521, Egypt.
authors.forename: D S
authors.initials: DS
authors.lastname: El-Kafrawy
authors.affiliation: University of Alexandria, Faculty of Pharmacy, Pharmaceutical Chemistry Department, Elmessalah, Alexandria 21521, Egypt.
authors.forename: M S
authors.initials: MS
authors.lastname: Mahrous
authors.affiliation: University of Alexandria, Faculty of Pharmacy, Pharmaceutical Chemistry Department, Elmessalah, Alexandria 21521, Egypt.
authors.forename: M M
authors.initials: MM
authors.lastname: Abdel-Khalek
authors.affiliation: University of Alexandria, Faculty of Pharmacy, Pharmaceutical Chemistry Department, Elmessalah, Alexandria 21521, Egypt.
authors.forename: A H
authors.initials: AH
authors.lastname: Abo-Gharam
mesh_terms: D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D020911:Ketorolac Tromethamine / Q000032:analysis*; D057230:Limit of Detection; D010656:Phenylephrine / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
chemical_list: D010656:Phenylephrine; D020911:Ketorolac Tromethamine
keywords: Derivative spectrophotometry; HPLC-DAD; HPTLC; Ketorolac tromethamine; Kétorolac trométhamine; Phenylephrine hydrochloride; Phényléphrine chlorhydrate; Ratio spectra; Spectres ratio; Spectrophotométrie dérivée