Development of sensitive benzofurazan‐based spectrometric methods for analysis of spectinomycin in vials and human biological samples

Abstract

Spectinomycin hydrochloride (SPEC) is an aminoglycoside antibiotic that has a broad spectrum against several bacterial strains of humans and veterinary infections.However, SPEC lacks a fluorophore or chromophore and this lack makes its analysis a challenge. Our study provides a simple, sensitive and low-cost spectrofluorimetric/spectrophotometric method based on the reaction between secondary amine groups and a benzofurazan reagent using borate buffer (pH 9.2). The yielding compound was measured fluorimetrically at 530 nm (excitation at 460 nm) with colorimetry at 410 nm. The calibration curves ranged from 30 to 400 ng ml −1 and from 0.2 to 6 μg ml −1 for spectrofluorimetric and spectrophotometric analyses, respectively. The limits of detection were calculated to be 4.15 ng ml −1 and 0.05 μg ml −1 for spectrofluorimetric and spectrophotometric processes, respectively.The ultra-affectability and high selectivity of the proposed method permitted analysis of SPEC in the dosage form and in human plasma samples, with good recoveries of about 101.19 and 97.11%, respectively, without any matrix interference. The proposed strategy was validated using International Conference on Harmonization standards and validated bioanalytically using USFDA recommendations.

Bio‐analytically fluorimetric method for estimation of ertapenem in real human plasma and commercial samples; application to pharmacokinetics study

“…NBD‐Cl is a fluorometric reagent that reacts with primary and secondary amine moieties to produce highly fluorescent products in alkaline medium. [ 1,3 ]…”

“…During the reaction of EPM with NBD‐Cl reagent, a bi‐product, 4‐hydroxy‐7‐nitrobenzo‐2‐oxa‐1,3‐diazole (NBD‐OH), was formed. [ 3,5 ] Therefore it was necessary to acidify the medium used in the reaction before dilution to decrease the fluorescence intensity of NBD‐OH without affecting the reaction product. [ 3,5 ] The fluorescence of NBD‐OH was quenched using 1 ± 0.25 ml of 2 M HCl without affecting the fluorescence of the reaction product.…”

“…4‐Chloro‐7‐nitrobenzofurazan (NBD‐Cl) is chemical fluorimetric derivative reagent used for analysis of primary and secondary amine moieties producing fluorescence products in alkaline medium. [ 1,3 ]…”

“…[1,2] In recent years, fluorimetric techniques have been the most applicable approach for estimation of varying pharmaceutical products in biomedical analysis because they are simple, sensitive, cost-effective, easily applied in a clinical laboratory, and selective. [1,[3][4][5][6] Varying methods have been reported for estimation of EPM such as spectrofluorimetry, [7] high performance liquid chromatography (HPLC) [8] and spectrophotometry. [9,10] Many drawbacks have been observed in the reported methods such as the use of highly expensive reagents, complicated equipment, and limited application.…”

“…4-Chloro-7-nitrobenzofurazan (NBD-Cl) is chemical fluorimetric derivative reagent used for analysis of primary and secondary amine moieties producing fluorescence products in alkaline medium. [1,3] The aim of this study was to establish a simple, selective, and sensitive method for analysis of ertapenem in real human plasma and pharmaceutical vials. Pharmacokinetic parameters such as maximum concentration (C max ) of EPM, half-life, and area under the curve were recorded without any matrix interference.…”