Liquid Chromatographic Determination of Amikacin Sulphate after Pre-Column Derivatization

Abstract

A novel high performance liquid chromatographic (HPLC) method with a pre-column derivatization reaction has been developed and validated. The method was used for the determination of the aminoglycoside antibiotic amikacin sulphate (AMK) in the presence of its synthetic precursor kanamycin sulphate in pure form and in different pharmaceutical preparations. The pre-column derivatization was based on Hantzsch condensation reaction and the obtained coloured products were separated using an isocratic reversed-phase high performance liquid chromatographic method. The separation was achieved on a Spherisorb C18 ODS2 (250 × 4.6 mm, 5 μm) column using a mobile phase composed of acetonitrile-0.1 M sodium acetate buffer (pH 5.0; 25:75, v/v). The column temperature was adjusted at 35 °C and the flow rate at 2 mL min(-1). The detection was carried out at 330 nm by using photo-diode array detector. Different conditions for the optimization of the derivatization reaction as well as for the HPLC measurement were studied. Moreover, AMK was subjected to forced degradation by oxidation, hydrolysis, photolysis and dry heat. Degradation products did not interfere with the assay, which can thus be considered selective and specific. The proposed method was validated for linearity, precision, accuracy, specificity and robustness. Also, it was used to check the purity of AMK in the presence of KAN (related impurity) at the pharmacopoeial limit (0.5%).

A rapid HPLC–DAD method for quantification of amikacin in pharmaceuticals and biological samples using pre-column derivatization with Hantzsch reagent

“…Derivatization of AMK drug with Hantzsch reagent was carried out by following a reported method [ 14 , 15 ] with some modification. Briefly, derivatization was carried by mixing 1.0 mL of AMK drug with 1.0 mL of freshly prepared Hantzsch reagent in to a 5.0 mL screw cap test tube then heated in water bath adjusted at 60 ºC for 15 min.…”

Simultaneous determination of ten aminoglycoside antibiotics in aquatic feeds by high‐performance liquid chromatography quadrupole‐orbitrap mass spectrometry with pass‐through cleanup

“…High‐performance liquid chromatography (HPLC) with ultraviolet (UV) or fluorescence (FL) detectors is a selective chemical method, which is widespread in the analysis of drug residues. Nevertheless, the molecular structures of AGs do not present chromophores or fluorophores, a pre‐ or post‐column derivatization step is required prior to HPLC analysis . To avoid the complicated procedures, mass spectrometry (MS) is an alternative for AGs analysis.…”

“…[7][8][9] High-performance liquid chromatography (HPLC)withultraviolet(UV)orfluorescence(FL)detectorsis aselectivechemicalmethod,whichiswidespreadintheanalysisofdrugresidues.Nevertheless,themolecularstructuresof AGs do not present chromophores or fluorophores, a pre-or post-column derivatization step is required prior to HPLC analysis. 10,11 Toavoidthecomplicatedprocedures,massspectrometry (MS) is an alternative for AGs analysis. Liquid chromatography-MS (LC-MS) and liquid chromatography tandemMS(LC-MS/MS)arewidelyusedmethodsforsingleor multi-AGsresiduesdetectionwithhighselectivityandsensitivity.…”

Analysis of Amikacin in Human Serum By UHPLC With Fluorescence Detector Using Chloro-Formate Reagent With Glycine

“…Normally serum fluid is selected for quantitative analysis of amikacin as it provides an accurate estimation of amikacin in body fluids due to its low protein binding property .1 At present, various methods for the determination of amikacin and other aminoglycosides have been developed using microbiological assay, high performance liquid chromatography (HPLC) with PDA. [4][5][6][7][8][9][10] and fluorescence, 11,12 Fluorimetric, 13 catalytic derivatization with HPLC, 14 spectrofluorimetery, 15 liquid chromatography-mass spectroscopy (LC-MS). 16 Amikacin sulfate molecule lacks structural properties necessary for production of signals compatible with photo diode array detector (PDA) (ultra violet and visible) for qualitative and quantitative analysis of drug molecule.…”