Measurement of ganciclovir concentration in human plasma by ultra-performance liquid chromatography-tandem mass spectrometry

Abstract

Background: Ganciclovir/valganciclovir plays an important role in the treatment and prevention of cytomegalovirus disease after organ transplantation.

Material and methods: We developed and validated a simple chromatographic method by ultra-performance liquid chromatography tandem mass spectrometry to measure plasma concentration of ganciclovir in human plasma. Chromatographic separation was achieved using an Acquity(®) UPLC(®) BEH™ (2.1×50mm id, 1.7μm) reverse-phase C18 column, with a water/methanol linear gradient containing ammonium acetate/formic acid at a 0.4mL/min flow rate. Ganciclovir and its internal standard (acyclovir) were detected by electrospray ionization mass spectrometry in positive ion multiple reaction monitoring mode.

Results: The limit of detection and quantification were 0.03 and 0.06mg/L, respectively, and linearity was observed between 0.06 and 30.0mg/L. Intra-day and day-to-day coefficients of variation and relative biases ranged from 3.6 to 5.4%, 4.2 to 6.2%, -2.6 to -1.1% and -4.0 to -2.8%, respectively. Recovery values were greater than 81.9%. Evaluation of the matrix effect showed ion suppression for ganciclovir and acyclovir. No carry-over was observed.

Conclusions: The validated method is useful for both therapeutic drug monitoring and pharmacokinetic studies. It could be applied to the daily clinical laboratory practice to measure the concentration of ganciclovir in human plasma.

Keywords: ACV; AUC; Acyclovir; CLSI; CMV; CV; Clinical and Laboratory Standards Institute; Cytomegalovirus; EMA; ESI; European Medicines Agency; GCV; Ganciclovir; HPLC; IFCC; International Federation of Clinical Chemistry and Laboratory Medicine; LLOD; LLOQ; MRM; MS; MS/MS; QC; S/N; SOT; Therapeutic drug monitoring; UPLC; UPLC–MS/MS; UV; VGCV; Valganciclovir; acyclovir; area under the curve; coefficient of variation; cytomegalovirus; electrospray ionization; ganciclovir; high-performance liquid chromatography; lower limit of detection; lower limit of quantification; m/z; mass spectrometer; mass-to-charge; multiple reaction monitoring; quality control; signal-to-noise; solid organ transplant; tandem mass spectrometry; ultra-performance liquid chromatography; ultraviolet; valganciclovir.