Development and Validation of an Ultra High-Performance Liquid Chromatographic Method with Ultraviolet Detection for Simultaneous Quantification of Filgrastim and Motixafortide in Human Plasma

Abstract

Multiple myeloma is a hematologic malignancy characterized by the uncontrolled proliferation of malignant plasma cells in the bone marrow. Filgrastim and motixafortide are used in combination to mobilize hematopoietic stem cells for collection prior to autologous stem cell transplantation, an important component of therapy for eligible patients with multiple myeloma. Accurate quantification of these agents in plasma is essential for therapeutic drug monitoring and bioequivalence studies. The objective of current work is to develop and validate a simple, accurate, and cost-effective ultra-high-performance liquid chromatographic method for the simultaneous estimation of filgrastim and motixafortide in human plasma. Protein precipitation was used for sample preparation. Chromatographic separation was achieved on a CHS C18 column with a mobile phase of 0.01 M ammonium acetate buffer pH 4.8 and acetonitrile (40:60, v/v) at a flow rate of 0.3 mL/min. Metformin was employed as the internal standard, and UV detection was performed at 220 nm. The retention times of filgrastim, motixafortide were 1.638 and 2.031 min,, respectively. The method was linear in the range of 5–200 ng/mL for filgrastim and 100–4000 ng/mL for motixafortide, with regression coefficients of 0.9993 and 0.9996, respectively. The lower limit of quantification was 5 ng/mL for filgrastim and 100 ng/mL for motixafortide. Accuracy values were within the range of 98.2% to 101.8%, and precision, evaluated as the percentage coefficient of variation, was below 2.0%. Mean recovery exceeded 95% for both analytes and the internal standard. The developed method is suitable for the simultaneous quantification of filgrastim and motixafortide in human plasm.