Development and application of a method for quantitative monitoring of Isepamicin concentration in human plasma based...

A rapid and quantitative analytical method for the detection of isepamicin (ISP) in human plasma was developed utilizing ultra-high performance liquid chromatography coupled with tandem triple quadrupole mass spectrometry (UHPLC-MS/MS). This method facilitates the monitoring of ISP plasma concentrations to examine distribution trends and the incidence of adverse reactions post-administration. Furthermore, the safety profile of ISP in clinical applications was evaluated from a therapeutic perspective by correlating ISP levels with patients' clinical recovery, thereby providing a reference framework for assessing ISP's safety.

The UHPLC-MS/MS analysis was conducted using a Waters XBridge BEH Amide column with a specifically formulated mobile phase. The gradient elution procedure, flow rate, and column temperature were optimized to achieve efficient sample separation and detection within a reduced time frame. Kanamycin was employed as an internal standard to ensure accurate quantification of ISP in plasma. Prior to analysis, plasma samples underwent a straightforward protein precipitation processing using acetonitrile. The method was validated in compliance with pertinent regulations to ensure its reliability. Specifically, the established method was validated in accordance with the relevant regulations outlined in the 2020 edition of the Chinese Pharmacopoeia and FDA bioassay methods. Patient samples receiving ISP were collected and subjected to comprehensive analysis.

The UHPLC-MS/MS method exhibited exceptional performance in quantifying ISP, demonstrating acceptable level of precision and accuracy. The relative standard deviation (RSD%) for inter-day precision ranged from 4.58 % to 6.40 %, while the RSD% for intra-day precision ranged from 1.19 % to 5.84 %, the relative error (RE%) values for both precision measures were within the range of 0.64 %-10.45 %. Additionally, the matrix effect observed for ISP was approximately 114 %. The RSD% for stability was less than 10.28 %. The experimental results conformed to the acceptance criteria for bioanalytical methods as recommended by the Chinese Pharmacopoeia and the FDA.

The UHPLC-MS/MS method is a robust tool for the determination of ISP, and the validated method has been successfully applied to monitor plasma drug concentrations in patients using ISP.