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. 2021 Oct 2;9(10):1379.
doi: 10.3390/biomedicines9101379.

A UHPLC-MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots

Federica Pigliasco  1 , Alessia Cafaro  1 , Raffaele Simeoli  2 , Sebastiano Barco  1 , Alberto Magnasco  3 , Maura Faraci  4 , Gino Tripodi  1 , Bianca Maria Goffredo  2 , Giuliana Cangemi  1
Affiliations

A UHPLC-MS/MS Method for Therapeutic Drug Monitoring of Aciclovir and Ganciclovir in Plasma and Dried Plasma Spots

Federica Pigliasco et al. Biomedicines. .

Abstract

The role of therapeutic drug monitoring (TDM) of valaciclovir (VA)/aciclovir (A) and valganciclovir/ganciclovir (VG/G) in critically ill patients is still a matter of debate. More data on the dose-concentration relationship might therefore be useful, especially in pediatrics where clinical practice is not adequately supported by robust PK studies. We developed and validated a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) micro-method to simultaneously quantify A and G from plasma and dried plasma spots (DPS). The method was based on rapid organic extraction from DPS and separation on a reversed-phase C-18 UHPLC column after addition of deuterated internal standards. Accurate analyte quantification using SRM detection was then obtained using a Thermo Fisher Quantiva triple-quadrupole MS coupled to an Ultimate 3000 UHPLC. It was validated following international (EMA) guidelines for bioanalytical method validation and was tested on samples from pediatric patients treated with A, VG, or G for cytomegalovirus infection following solid organ or hematopoietic stem cell transplantation. Concentrations obtained from plasma and DPS were compared using Passing-Bablok and Bland-Altman statistical tests. The assay was linear over wide concentration ranges (0.01-20 mg/L) in both plasma and DPS for A and G, suitable for the expected therapeutic ranges for both Cmin and Cmax, accurate, and reproducible in the absence of matrix effects. The results obtained from plasma and DPS were comparable. Using an LC-MS/MS method allowed us to obtain a very specific, sensitive, and rapid quantification of these antiviral drugs starting from very low volumes (50 μL) of plasma samples and DPS. The stability of analytes for at least 30 days allows for cost-effective shipment and storage at room temperature. Our method is suitable for TDM and could be helpful for improving knowledge on PK/PD targets of antivirals in critically ill pediatric patients.

Keywords: LC-MS/MS; aciclovir; dried plasma spot; ganciclovir; therapeutic drug monitoring; valaciclovir; valganciclovir.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The mean calibration curves (9 point calibration curve) of ganciclovir and aciclovir ranging from 0.01 to 20 mg/L in plasma (panel a) and in DPS (panel b).
Figure 2
Figure 2
Chromatograms obtained: a calibrator at the LLOQ in plasma (panel 1.a, aciclovir; panel 2.a, ganciclovir); deuterated internal standards (panel 3.a, ganciclovir-d5) and a calibrator at the LLOQ in DPS (panel 1.b, aciclovir; panel 2.b, ganciclovir); deuterated internal standards (panel 3.b, ganciclovir-d5). RT, retention time; AA, automatic area; SN, signal-to-noise ratio; NL, normalized level.
Figure 3
Figure 3
Passing–Bablok correlation plots between aciclovir (A) and ganciclovir (B) concentrations extracted from plasma. Aciclovir/ganciclovir concentrations (mg/L) measured from plasma are represented on the x-axis and aciclovir/ganciclovir concentrations (mg/L) from DPS on the y-axis. Thick line, regression line; Thin line, identity line; Dashed line, confidence interval for the regression line.
Figure 4
Figure 4
Bland–Altman plots between aciclovir (A) and ganciclovir (B) concentrations extracted from plasma. On the x-axis are the average of the plasma concentration and DPS concentration of aciclovir/ganciclovir (mg/L) measured from plasma and DPS, and the y-axis represents the difference between the plasma concentrations and DPS concentrations of aciclovir/ganciclovir (mg/L).
See this image and copyright information in PMC

References

    1. Abdul-Aziz M.H., Alffenaar J.W.C., Bassetti M., Bracht H., Dimopoulos G., Marriott D., Neely M.N., Paiva J.A., Pea F., Sjovall F., et al. Antimicrobial therapeutic drug monitoring in critically ill adult patients: A Position Paper#. Intensive Care Med. 2020;46:1127–1153. doi: 10.1007/s00134-020-06050-1. - DOI - PMC - PubMed
    1. Roberts J.A., Abdul-Aziz M.H., Lipman J., Mouton J.W., Vinks A.A., Felton T.W., Hope W.W., Farkas A., Neely M.N., Schentag J.J., et al. Individualised antibiotic dosing for patients who are critically ill: Challenges and potential solutions. Lancet Infect. Dis. 2014;14:498–509. doi: 10.1016/S1473-3099(14)70036-2. - DOI - PMC - PubMed
    1. Tabah A., de Waele J., Lipman J., Zahar J.R., Cotta M.O., Barton G., Timsit J.F., Roberts J.A. The ADMIN-ICU survey: A survey on antimicrobial dosing and monitoring in ICUs. J. Antimicrob. Chemother. 2015;70:2671–2677. doi: 10.1093/jac/dkv165. - DOI - PubMed
    1. Wong G., Brinkman A., Benefield R.J., Carlier M., De Waele J.J., El Helali N., Frey O., Harbarth S., Huttner A., McWhinney B., et al. An international, multicentre survey of β-lactam antibiotic therapeutic drug monitoring practice in intensive care units. J. Antimicrob. Chemother. 2014;69:1416–1423. doi: 10.1093/jac/dkt523. - DOI - PubMed
    1. Gustinetti G., Cangemi G., Bandettini R., Castagnola E. Pharmacokinetic/pharmacodynamic parameters for treatment optimization of infection due to antibiotic resistant bacteria: A summary for practical purposes in children and adults. J. Chemother. 2018;30:65–81. doi: 10.1080/1120009X.2017.1377909. - DOI - PubMed