ESTIMATION OF GILTERITINIB IN HUMAN PLASMA BY LC-ESI-MS/MS: METHOD DEVELOPMENT AND VALIDATION

Ms. Sharma Supriya Jitendra*, Ms. Rashmi R. Shukla, Dr. Tulsidas Mishra, Dr. Pinak Patel, Dr. Pritesh Contractor

ABSTRACT

A simple, rapid and sensitive bioanalytical method developed to validate the quantity of Gilteritinib drug in human
plasma. The development of method was done with the help of LC-ESI-MS/MS system. This method employs
binary normal phase phase liquid chromatography-tandem mass spectroscopy operating in Multiple Reaction
Monitoring (MRM) mode for detection. This method uses Electron spray ionization source and operates in positive
mode. Solid phase extraction method was used to prepare samples, yielding high recovery rates for both Gilteritinib
and its internal standard (ISTD). Chromatographic separation was achieved on column Kinetex, 2.6 μm Biphenyl
(50mm × 4.6mm) with a mobile phase comprising Pump-A: 0.1% Formic acid in 10Mm Ammonium format and
Pump B: Organic mixture (Methanol: Acetonitrile, 50:50 v/v) at yellow monochromatic light under Ambient
temperatuare (25 ±5°C). The analyte and ISTD were eluted at approximately 2.7 minutes with total runtime of 5
minutes. Sample prepration was done by Solid phase extraction and this method yelds recover 75.91% for drug
and 77.72% for ISTD respectively. The method demonstrated excellent linearity over a dynamic range of 0.200
ng/mL to 130.000 ng/mL, with correlation coefficients consistently within the acceptable range. The regression
analysis were found in range of 0.9989. The accuracy of the method met regulatory criteria, within acceptance
limit of ±20 at lower limit of quantification(LLOQ) and ±15% at other concentrations. The method was validated
following ICH M10, ANVISA and USFDA guidelines, and confirming its reliability for bioanalytical application in
pharmacokinetic and clinical studies.

Keywords: Electron spray ionization, Bioanalytical method development, Gilteritinib, Solid-phase Extraction, Validation, LC-ESI-MS/MS.