Sensitive determination of gentamicin in plasma using ion-exchange solid-phase extraction followed by UHPLC-MS/MS analysis

Ana Laura Anibaletto Dos Santos; Anne Caroline Cezimbra da Silva; Lilian de Lima Feltraco Lizot; Anelise Schneider; Roberta Zilles Hahn; Yasmin Fazenda Meireles; Lidiane Riva Pagnussat; Julia Livia Nonnenmacher; Siomara Regina Hahn; Rafael Linden

doi:10.1016/j.plabm.2021.e00246

Sensitive determination of gentamicin in plasma using ion-exchange solid-phase extraction followed by UHPLC-MS/MS analysis

Pract Lab Med. 2021 Jul 16:26:e00246. doi: 10.1016/j.plabm.2021.e00246. eCollection 2021 Aug.

Affiliations

¹ Analytical Toxicology Laboratory, Universidade Feevale, Novo Hamburgo, RS, Brazil.
² Graduate Program on Toxicology and Analytical Toxicology, Universidade Feevale, Brazil.
³ Hospital São Vicente de Paulo, Passo Fundo, RS, Brazil.
⁴ Multidisciplinary Residency Program, Universidade de Passo Fundo, RS, Brazil.
⁵ College of Pharmacy, Universidade de Passo Fundo, RS, Brazil.

Abstract

Background: Therapeutic drug monitoring (TDM) of gentamicin sulfate (GEN) is usually recommended, particularly in critical patients. Only a few reports had described the determination of GEN in plasma or plasma using LC-MS/MS.

Objective: This study aimed to develop and validate a sensitive ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) assay for the quantification of GEN in small volumes of human plasma.

Results: The use of a very low concentration of the ion-pairing agent HFBA allowed significant retention of the very polar GEN forms in a reversed phase UHPLC column. The solid-phase extraction (SPE) procedure allowed clean extracts, with no interferences detected in blank samples, and high sensitivity. The assay was linear on the range of 0.2-40 mg L^-1 of GEN complex. The combined GEN complex had inter-assay CV of 8.8-10.0%, intra-assay CV of 10.2-11.0%, and accuracy of 96.8-104.0%. The assay was applied to 17 clinical samples obtained from neonate patients. Measured concentrations were in the range of 0.15-3.57 mg L^-1 for GEN C1, 0.12-3.55 mg L^-1 for GEN C1a, 0.20-5.77 mg L^-1 for GEN C2, and 0.47-12.88 mg L^-1 for the GEN complex, all within the linear range of the assay.

Conclusion: A sensitive assay for the quantification of gentamicin in plasma using anion-exchange SPE and UHPLC-MS/MS was validated. The assay can be used for TDM of gentamicin, particularly in centers with access to proper instrumentation and with a low demand for gentamicin measurements, where immunoassays are not cost-effective.

Keywords: Gentamicin; Neonates; Solid-phase extraction; Therapeutic drug monitoring; UHPLC-MS/MS.