Liquid chromatography–tandem mass spectrometry for the simultaneous quantitation of ceftriaxone, metronidazole and hydroxymetronidazole in plasma from seriously ill, severely malnourished children

Wellcome Open Research

article

Martin Ongas¹  Joseph Standing³  Bernhards Ogutu¹  Joseph Waichungo⁵  James A. Berkley⁵  Karin Kipper⁴ 

[1] Center for Research in Therapeutic Sciences, Strathmore University;KEMRI-Centre for Clinical Research;Inflammation, Infection and Rheumatology Section, UCL Great Ormond Street Institute of Child Health;Paediatric Infectious Diseases Research Group, Institute for Infection and Immunity, St. George's, University of London;KEMRI-Wellcome Trust Research Programme;Centre for Tropical Medicine & Global Health, Nuffield Department of Medicine, University of Oxford;The Childhood Acute Illness & Nutrition;Analytical Services International, St George’s University of London;Institute of Chemistry, University of Tartu

关键词: LC-MS/MS;    ceftriaxone;    metronidazole;    complicated severe acute malnutrition;    ultrafiltration;    protein binding;   

DOI  :  10.12688/wellcomeopenres.11728.2

学科分类：内科医学

来源: Wellcome

【 摘 要 】

We have developed and validated a novel, sensitive, selective and reproducible reversed-phase high-performance liquid chromatography method coupled with electrospray ionization mass spectrometry (HPLC–ESI-MS/MS) for the simultaneous quantitation of ceftriaxone (CEF), metronidazole (MET) and hydroxymetronidazole (MET-OH) from only 50 µL of human plasma, and unbound CEF from 25 µL plasma ultra-filtrate to evaluate the effect of protein binding. Cefuroxime axetil (CEFU) was used as an internal standard (IS). The analytes were extracted by a protein precipitation procedure with acetonitrile and separated on a reversed-phase Polaris 5 C18-Analytical column using a mobile phase composed of acetonitrile containing 0.1% (v/v) formic acid and 10 mM aqueous ammonium formate pH 2.5, delivered at a flow-rate of 300 µL/min. Multiple reaction monitoring was performed in the positive ion mode using the transitionsm/z555.1→m/z396.0 (CEF),m/z172.2→m/z 128.2 (MET),m/z188.0→m/z125.9 (MET-OH) andm/z528.1→m/z 364.0 (CEFU) to quantify the drugs. Calibration curves in spiked plasma and ultra-filtrate were linear (r2 ≥ 0.9948) from 0.4–300 µg/mL for CEF, 0.05–50 µg/mL for MET and 0.02 – 30 µg/mL for MET-OH. The intra- and inter- assay precisions were less than 9% and the mean extraction recoveries were 94.0% (CEF), 98.2% (MET), 99.6% (MET-OH) and 104.6% (CEF in ultra-filtrate); the recoveries for the IS were 93.8% (in plasma) and 97.6% (in ultra-filtrate). The validated method was successfully applied to a pharmacokinetic study of CEF, MET and MET-OH in hospitalized children with complicated severe acute malnutrition following an oral administration of MET and intravenous administration of CEF over the course of 72 hours.

【 授权许可】

CC BY