Affiliation:
1. Doshisha Women’s College of Liberal Arts, Faculty of Pharmaceutical Sciences, Department of Analytical Chemistry , 97-1 Minamihokodate Kodo Kyotanabe , Kyoto 610-0395, Japan
2. Institute of Microbial Chemistry (BIKAKEN) , 3-14-23, Kamiosaki, Shinagawa-ku , Tokyo 141-0021, Japan
3. Antimicrobial Resistance Research Center, National Institute of Infectious Diseases , Tokyo 189-0002, Japan
4. Pharmaceutical Reference Standards Center, Pharmaceutical and Medical Device Regulatory Science Society of Japan , Osaka 541-0046, Japan
Abstract
Abstract
Background
Through the recent development of analytical technology, antibiotics quantification in the Japanese Pharmacopoeia (JP) has changed from traditional microbiological assays to physicochemical methods with high specificity and precision. However, for several multicomponent antibiotics without typical UV absorption, potency cannot be directly determined using instrumental methods such as high-performance liquid chromatography; therefore, traditional microbiological assays are still used. Gentamicin sulfate (GmS), which consists of three major components, C1, C1a, and C2, is such a typical antibiotic, and its antimicrobial potency continues to be assayed using microbiological methods in JP monographs. Introduction of a physicochemical assay for GmS is needed to help ensure its quality and quantity.
Objective
This study aimed to develop quality control measures for GmS that could be complementary to quantitative assays and purity tests specified in the JP.
Methods
For each gentamicin C component (C1, C2, and C1a), theoretical potencies were determined based on the quantitative relationship between purity and potency, as measured by quantitative 1H NMR and microbiological assays, respectively. Two lots of the JP reference standard (RS) were used as test samples, with the contents of each component and impurity (sisomicin and garamine) being determined using hydrophilic interaction liquid chromatography–tandem mass spectrometry (HILIC-MS/MS).
Results
The ratios of theoretical potency for C1, C2, and C1a were 1.00, 1.21, and 1.80, respectively. The potencies of the GmS JP RSs, which were estimated based on the contents and theoretical potency of each C component, corresponded well with those determined through microbiological assays. Marked differences in impurities (%) between the two RS lots were highlighted by quantifying sisomicin and garamine.
Conclusions
The developed analytical procedure enabled the characterization of two different JP RSs in terms of content ratio, potencies, and impurities.
Highlights
Novel analytical procedures useful for routine quality control of GmS were developed using HILIC-MS/MS.
Funder
Japan Agency for Medical Research and Development
Publisher
Oxford University Press (OUP)
Subject
Pharmacology,Agronomy and Crop Science,Environmental Chemistry,Food Science,Analytical Chemistry