Analysis of kanamycin A in human plasma and in oral dosage form by derivatization with 1-naphthyl isothiocyanate and high-performance liquid chromatography

Abstract

A simple and sensitive HPLC method has been developed for trace determination of kanamycin A by derivatization. Plasma proteins are precipitated by acetonitrile and chemical derivatization is performed on the supernatant containing kanamycin A with 1-naphthyl isothiocyanate in pyridine at 70 degrees C. After the derivatization reaction, a methylamine/acetonitrile solution was added to the reaction mixture to eliminate the excess of derivatizing agent and shorten the analysis time. The resulting derivative was separated using a Lichrocart Purospher STAR RP-18e column and water/methanol (33:67, v/v) as a mobile phase (detection at 230 nm). Optimization conditions for the derivatization of kanamycin A were investigated by HPLC. The linear range for the quantitation of kanamycin A in spiked plasma was over 1.2-40 microg/mL; the detection limit (signal to noise ratio = 3; injection volume, 10 microL) was about 0.3 microg/mL. The relative standard deviation was less than 2.9% for intra-day assay (n = 6) and inter-day assay (n = 6) and relative recoveries were found to be greater than 98%. Preliminary application of the method for monitoring kanamycin A in humans upon intramuscular injection of the injection product demonstrated the usefulness of the assay for clinical studies. The proposed method can also be used to analyze the compound in pharmaceutical formulations.

Determination of kanamycin A, amikacin and tobramycin residues in milk by capillary zone electrophoresis with post-column derivatization and laser-induced fluorescence detection

“…For the proposed method, the LODs of amikacin, kanamycin A and tobramycin for standard solution and milk sample were 8 × 10 −6 and 1 × 10 −5 g/L, 7 × 10 −6 and 7 × 10 −6 g/L, and 2 × 10 −5 and 3 × 10 −5 g/L, respectively, which were better than those of nonfluorescence detection methods of CE, such as UV [12,15,17] and ED [20,21,23] detection, but inferior to those of the CE-LIF with pre-column derivatization method [25]. The LODs of the proposed method were also better than those of LC-UV for the derivatized AGs and LCchemiluminescence (CL) [6,7]. It was difficult to compare with LC-MS because of the lack of the reported works on the three AGs [8,9].…”

“…Chromatographic methods have been used in the analysis of AGs [2], including thin-layer chromatography (TLC) [3], gas chromatography (GC) [4], liquid chromatography (LC) [5][6][7] and liquid chromatography-mass spectrometry (LC-MS) [8,9]. LC and LC-MS are the main analytical techniques of AGs because of their capabilities of separation, identification and quantification.…”

Development and validation of a capillary electrophoresis method with capacitively coupled contactless conductivity detection (CE‐C4D) for the analysis of amikacin and its related substances

“…Several methods have been applied for determination of amikacin in various matrices including microbiological assays , radioenzymatic assays , and immunoassays . Chromatographic methods have been used in the analysis of AGs including TLC , GC , and LC . LC was combined with pre‐ and postcolumn derivatization with O ‐phthalaldehyde , precolumn derivatization with 1‐fluoro‐2,4‐dinitrobenzene , or trinitrobenzenesulphonic acid .…”

LC method for the analysis of kanamycin residue in swine tissues using derivatization with 9‐fluorenylmethyl chloroformate

“…Numerous methods for the detection of Kan in various biological matrixes have been reported in literature. These methods involved immunoassays 5–7, CE 8, 9, GC‐MS 10, LC 11–13, and LC‐MS/MS 14–16. Immunoassay is generally used as the screening method because of its susceptibility to environmental factors, and the suspected noncompliant results detected by immunoassay should be further validated by instrumental methods.…”

“…However, the instruments involved in the two methods are not routinely available in most laboratories because of their low usage frequency or high cost. Because of its convenience and low demand for instrument, HPLC method is suitable as a routine method to analyze Kan in different samples 11–13. Owing to the lack of strong chromophore in molecule, Kan has been derivatized with phenylisocyanate 11 or 1‐naphthyl isothiocyanate 12 and detected by UV detector.…”