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

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.