Validation and Application of a Dried Blood Spot Ceftriaxone Assay

Abstract

dDried blood spot (DBS) antibiotic assays can facilitate pharmacokinetic/pharmacodynamic (PK/PD) studies in situations where venous blood sampling is logistically and/or ethically problematic. In this study, we aimed to develop, validate, and apply a DBS ceftriaxone assay. A liquid chromatography-tandem mass spectroscopy (LC-MS/MS) DBS ceftriaxone assay was assessed for matrix effects, process efficiency, recovery, variability, and limits of quantification (LOQ) and detection (LOD). The effects of hematocrit, protein binding, red cell partitioning, and chad positioning were evaluated, and thermal stability was assessed. Plasma, DBS, and cell pellet ceftriaxone concentrations in 10 healthy adults were compared, and plasma concentration-time profiles of DBS and plasma ceftriaxone were incorporated into population PK models. The LOQ and LOD for ceftriaxone in DBS were 0.14 mg/liter and 0.05 mg/liter, respectively. Adjusting for hematocrit, red cell partitioning, and relative recovery, DBS-predicted plasma concentrations were comparable to measured plasma concentrations (r > 0.95, P < 0.0001), and Bland-Altman plots showed no significant bias. The final population PK estimates of clearance, volume of distribution, and time above threshold MICs for measured and DBS-predicted plasma concentrations were similar. At 35°C, 21°C, 4°C, ؊20°C, and ؊80°C, ceftriaxone retained >95% initial concentrations in DBS for 14 h, 35 h, 30 days, 21 weeks, and >11 months, respectively. The present DBS ceftriaxone assay is robust and can be used as a surrogate for plasma concentrations to provide valid PK and PK/PD data in a variety of clinical situations, including in studies of young children and of those in remote or resource-poor settings.

Validation and Application of a Dried Blood Spot Assay for Biofilm-Active Antibiotics Commonly Used for Treatment of Prosthetic Implant Infections

“…Our detailed approach to laboratory-based and clinical validation of DBSs for PK studies in this and a recent study (32) supports the use of this method for research and therapeutic drug monitoring in a variety of health care settings. Staff training is straightforward, disposables are relatively inexpensive, and the essential equipment required at a field site is modest.…”

Penicillin Dried Blood Spot Assay for Use in Patients Receiving Intramuscular Benzathine Penicillin G and Other Penicillin Preparations To Prevent Rheumatic Fever

“…Chromatographic data were processed using LAB Solution (Version 5.56; Shimadzu, Japan). Matrix effects (ion suppression/enhancement), absolute recovery, and process efficiency were determined at 0.1, 1, 10, and 100 mg/liter using our established method (13,14). The lower limit of quantification (LOQ) and lower limit of detection (LOD) were determined at signal-to-noise ratios of 10:1 and 3:1, respectively.…”

“…We have previously demonstrated the accuracy and feasibility of this method for assay of other antibiotics (13,14). Very low (20-l) volumes of whole blood can be collected from finger or heel prick samples and impregnated into filter paper, which is then stored in foil bags without the need for processing of plasma and immediate frozen storage.…”

A Critical Review of Analytical Methods for Determination of Ceftriaxone Sodium

“…Blood [49] HPLC-UV: High-performance liquid chromatography with detection by ultraviolet; HPLC-MS: High-performance liquid chromatography coupled with mass spectrometry detection; UPLC-MS/MS: Ultra-performance liquid chromatography coupled with sequential mass spectrometry; HPLC-MS/MS: High-performance liquid chromatography coupled with sequential mass spectrometry and UPLC-UV: Ultra-performance liquid chromatography with detection by ultraviolet; ND: not declared. Absorption spectroscopy VIS Dilution of 250 mg of sample in water, after filtration was diluted with distilled water and made up to 100 mL.…”