A Validated Stability-Indicating HPLC Method for Simultaneous Determination of Amoxicillin and Enrofloxacin Combination in an Injectable Suspension

Abstract

The combination of amoxicillin and enrofloxacin is a well-known mixture of veterinary drugs; it is used for the treatment of Gram-positive and Gram-negative bacteria. In the scientific literature, there is no high-performance liquid chromatography (HPLC)-UV method for the simultaneous determination of this combination. The objective of this work is to develop and validate an HPLC method for the determination of this combination. In this regard, a new, simple and efficient reversed-phase HPLC method for simultaneous qualitative and quantitative determination of amoxicillin and enrofloxacin, in an injectable preparation with a mixture of inactive excipients, has been developed and validated. The HPLC separation method was performed using a reversed-phase (RP)-C18e (250 mm × 4.0 mm, 5 μm) column at room temperature, with a gradient mobile phase of acetonitrile and phosphate buffer containing methanol at pH 5.0, a flow rate of 0.8 mL/min and ultraviolet detection at 267 nm. This method was validated in accordance with the Food and Drug Administration (FDA) and the International Conference on Harmonisation (ICH) guidelines and showed excellent linearity, accuracy, precision, specificity, robustness, ruggedness, and system suitability results within the acceptance criteria. A stability-indicating study was also carried out and indicated that this method can also be used for purity and degradation evaluation of these formulations.

Development and validation of RP-HPLC method for determination of amoxicillin residues and application to NICOMAC coating machine

“…2 AMO is official in British Pharmacopeia (BP), 1 European Pharmacopeia (EP) 3 and United States Pharmacopeia (USP) 4 , they include HPLC method for its determination. It is still a limited number of analytical methods that are reported for the determination of AMO including kinetics degradation, [5][6][7] spectrophotometric, [8][9][10][11][12][13] UHPLC UPLC and mass spectrometry, [14][15][16][17][18][19] thin layer chromatography (TLC), [20][21][22] capillary electrophoresis, [23][24][25][26] high performance liquid chromatography (HPLC), [27][28][29][30][31] in vitro dissolution studies, [32][33][34][35][36] amoxicillin residues in animal tissues using SPE-LC, 37 SPE-cation exchange, 38 in eggs using HPLC-FLD, 39 or HPLC-MS 40 and in commercial meat and milk samples 41 using HPLC-FLD. According to the best of our knowledge there is no validated method for the determination of amoxicillin residues and application to cleaning machine in pharmaceutical industries.…”

Stability-indicating HPLC-DAD method for the simultaneous determination of fluoroquinolone in combination with a non-steroidal anti-inflammatory drug in pharmaceutical formulation

“…Another stability-indicating study with ENRO in combination with amoxicillin found no degradation for ENRO. There was no evidence of degradation of the drug when exposed to acidic (HCl 0.1 N for 4 h; room temperature), alkaline (NaOH 0.1 N; 65 °C for 7 days), oxidative (H 2 O 2 0.3% for 7 days; room temperature), thermal (70 °C for 3 weeks), and photo (UV light for 3 days) stress conditions (Batrawi, Wahdan, Al-Rimawi, 2017).…”

“…Several analytical methods have been described for the estimation of ENRO or PIRO alone or combined with other drugs in pharmaceutical formulations. Studies have employed high-performance liquid chromatography (HPLC), thin-layer chromatography (TLC), UV spectrophotometry, spectrofluorimetry, infrared spectroscopy, potentiometry, voltammetry, phosphorimetry, or chemiluminescence (Batrawi, Wahdan, Al-Rimawi, 2017;Starek et al, 2009;Kormosh, Hunka, Bazel, 2011;Ulu, 2009;Golovnev, Vasiliev, Kiriki, 2012;Khaled et al, 2012;Ensaifi, Khayamian, Taei, 2009;Souza et al, 2013;Pulgarin, Molina, Munoz, 2011).…”

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“…The following quantitative procedures of penicillin determination are described: using potentiometry titration and ionometry, spectrophotometry, extraction photometry, voltammetry and polarography, micelle electrokinetic capillary and paper chromatography, chemiluminescence, electro-phoresis, and kinetic analysis methods [3][4][5][6][7][8][9][10][11][12][13][14] . A new procedure for the quantitative determination of Mezl in the Baypen® preparation by the method of back iodometric titration using potassium hydroperoxymonosulfate (KHSO 5 ) as an analytical reagent was developed.…”