Development and validation of a volumetric absorptive microsampling- liquid chromatography mass spectrometry method for the analysis of cefepime in human whole blood: Application to pediatric pharmacokinetic study

Abstract

Cefepime is a fourth-generation cephalosporin antibiotic with an extended spectrum of activity against many Gram-positive and Gram-negative bacteria. There is a growing need to develop sensitive, small volume assays, along with less invasive sample collection to facilitate pediatric pharmacokinetic clinical trials and therapeutic drug monitoring. The volumetric absorptive microsampling (VAMS™) approach provides an accurate and precise collection of a fixed volume of blood (10 μL), reducing or eliminating the volumetric blood hematocrit assay-bias associated with the dried blood spotting technique. We developed a high-performance liquid chromatographic method with tandem mass spectrometry detection for quantification of cefepime. Sample extraction from VAMS™ devices, followed by reversed-phase chromatographic separation and selective detection using tandem mass spectrometry with a 4 minute runtime per sample was employed. Standard curves were linear between 0.1 -100 μg/mL for cefepime. Intra

Therapeutic Drug Monitoring Is a Feasible Tool to Personalize Drug Administration in Neonates Using New Techniques: An Overview on the Pharmacokinetics and Pharmacodynamics in Neonatal Age

“…To date, VAMS is still a novel microsampling method and is not yet routinely used in neonates. Since first data published in 2014 [40], several drugs have been monitored through VAMS such as antiepileptics, anthelmintic and immunosuppressants [41][42][43][44]. VAMS can be utilized to collect blood samples from a single finger or heel prick, allowing to obtain accurate volumes, without hematocrit's influence.…”

Microsampling Assays for Pharmacokinetic Analysis and Therapeutic Drug Monitoring of Antimicrobial Drugs in Children: A Critical Review

“…The VAMS method was applied for quantifying cefepime in 7 clinical samples obtained from 1 pediatric subject. 23 The results demonstrated that the range and sensitivity of the assay were suitable for this VAMS-based LC-MS/MS assay. Piperacillin, tazobactam, meropenem, ceftazidime, and linezolid were simultaneously analyzed in 11 samples obtained from 7 pediatric patients.…”

“…Table 2 provides an overview of the validation results of recently published antimicrobial VAMS-based assays. The interassay accuracy and precision of VAMS-based assays for quantifying cefepime, 23 piperacillin, 17 tazobactam, 17 meropenem, 17 ceftazidime, 17 and linezolid 17 were acceptable. These VAMS assays have been validated and applied to clinical sample analysis.…”

“…There are validated methods for quantifying antimicrobial drugs by VAMS-based LC-MS/MS methods, which have been developed, validated, and implemented for analyzing clinical samples. 17,23 Sample Analysis LC-MS/MS is a common analytical technique that is used for quantifying antibiotics in human matrices, typically in plasma or serum samples. 13 With the advancement in technology, more advanced instruments have been developed that readily detect and quantify very low concentrations of analytes with little sample volume, which make DBS and VAMS suitable for quantitative analysis.…”

“…It has been demonstrated that 1 hour of drying at RT is sufficient for methods using 10 mL VAMS. 17,23 The linearity is evaluated for defining a wide dynamic range that includes the expected therapeutic concentration range of antimicrobials. The upper limit of quantitation should be sufficiently high to limit the number of samples that require dilution.…”