Validated HPLC method for ceftriaxone from dried blood spots for pharmacokinetic studies and therapeutic drug monitoring in neonatal population.

Background: Pharmacokinetic evaluation is essential for the precise dosing of ceftriaxone in neonates. There is a need for developing a sensitive, affordable and convenient analytical method that can estimate ceftriaxone from dried blood spot (DBS) samples of neonates. Method: An HPLC-UV method was developed and validated as per ICH M10 for ceftriaxone from DBS and plasma using an Inertsil-ODS-3V column with gradient elution. DBS samples were extracted with methanol. Clinical validation was performed using neonatal samples. Results: The developed plasma- and DBS-based-HPLC method were linear from 2-700 μg/ml and 2-500 μg/ml, respectively, for ceftriaxone. Bland-Altman analysis indicated a strong interconvertibility between the plasma and DBS assays. Conclusion: Observed concentrations in clinical samples were comparable to the predicted concentrations, proving the clinical validity of the method.