A novel high throughput 96‐well‐based fluorimetric method to measure amikacin in pharmaceutical formulations: development using response surface methodology. (10th April 2022)

• Record Type: Journal Article
• Authors: Maxwell, Amala; Mary, Elizabeth; Ghate, Vivek; Aranjani, Jesil; Lewis, Shaila
• Is Part Of: Luminescence. Volume 37:Number 6(2022)
• Page Start: 930
• Page End: 943
• Publication Date: 2022-04-10
• Subjects: amikacin -- pharmaceutical formulation -- response surface methodology -- spectrofluorimetry; Luminescence -- Periodicals; Bioluminescence -- Periodicals; Chemiluminescence -- Periodicals; Luminescence -- Periodicals; 535.35
• DOI: 10.1002/bio.4238
• Languages: English
• ISSNs: 1522-7235

Abstract

Abstract: An aminoglycoside antibiotic, amikacin, is used to treat severe and recurring bacterial infections. Due to the absence of a chromophore, however, amikacin must be extensively derivatized before being quantified, both in analytical and bioanalytical samples. In this study, for the first time, we developed a simple and sensitive method for measuring amikacin sulfate using spectrofluorimetry with a 96‐well plate reader, based on the design of the experiment's approach. To develop a robust and reproducible spectrofluorimetric method, the influence of essential attributes, namely pH of the buffer, heating temperature, and concentration of reagents, were evaluated using univariate analysis followed by multivariate analysis (central composite design). International Conference of Harmonization guidelines were used to validate the optimized method. The developed technique is linear from 1.9 to 10 μg/ml with a regression coefficient of 0.9991. The detection and quantification limits were 0.649 μg/ml and 1.9 μg/ml, respectively. For the developed method, both intraday and interday precision (%RSD) were less than 5%. Using the method, amikacin concentrations were quantified in prepared amikacin liposomes and commercial formulations of Amicin®. The developed method greatly reduces sample volume and is a rapid, high throughput microplate‐based fluorescence approach for the convenient and cost‐effective measurement of amikacin in pharmaceutical formulations. In comparison with previously published approaches, the suggested method allowed for quick analysis of a high number of samples in a short amount of time (96 samples in 125 sec), resulting in an average duration of analysis of 1.3 sec per sample. Abstract : Optimized a simple, rapid, convenient, sensitive and cost‐effective spectrofluorimetric method for amikacin by an experimental design approach using 96‐well plate fluorescence microplate reader. Validated the spectrofluorimetric method according to ICH Q2 (R1) and USFDA guidelines by analysing, linearity, sensitivity, specificity, accuracy, precision, bench‐top stability and established the stability of the analytical method at room temperature, refrigeration and dark. Quantified amikacin concentration in formulated nano carriers and compared by estimating amikacin concentration in the marketed amikacin injection.