Development and validation of a UPLC-PDA method for quantifying ceftazidime in dried blood spots
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High-throughput and Comprehensive Drug Surveillance Using Multisegment Injection-Capillary Electrophoresis-Mass Spectrometry
Published on: April 23, 2019
Jianmei Lv1, Qiping Wu1, Sanwang Li2
1Division of Biopharmaceutics and Pharmacokinetics, Xiangya School of Pharmaceutical Sciences, Central South University, Changsha 410013, China.
Abstract:
Bacterial infection is a leading cause of neonatal death. Ceftazidime, commonly used for neonatal infections, is often used off-label. Blood sampling limits pharmacokinetic (PK) studies in neonatal patients. The dried blood spots (DBS) are a potential matrix for microsampling. Herein, we describe an ultra-performance liquid chromatography with a photodiode array (UPLC-PDA) to determine ceftazidime in DBS from neonatal patients in support of pharmacokinetic studies. The Capitainer® device-based DBS samples containing 10 µL blood were extracted in 70% methanol/water (v/v) with acetaminophen as the internal standard (IS). The extraction process was carried out at 20 °C using a block bath shaker at 1000 rpm for 30 min. The extracted ceftazidime was subsequently eluted through an Acquity UPLC HSS T3 column (2.1 × 50 mm, 1.8 µm). Elution was achieved using a water (containing 0.1% trifluoroacetic acid)/acetonitrile linear gradient at a flow rate of 0.5 mL/min, and the analytical time was 3.2 min. The PDA detection wavelength was set at 259 nm. The method underwent thorough validation following the recommendation of the European Bioanalysis Forum (EBF) and the bioanalytical guideline established by the European Medicines Agency (EMA). No interfering peaks were detected at the retention times of ceftazidime and IS. The ceftazidime exhibited a quantification range spanning from 0.5 to 200 µg/mL, and the assay demonstrated good accuracy (intra/inter-assay ranging from 90.1% to 104.8%) and precision (intra/inter-assay coefficient of variations ranging from 4.8% to 11.7%). The method's applicability was demonstrated by analyzing clinical DBS samples collected from neonatal patients.
Insights
This study developed a new method using dried blood spots (DBS) to measure ceftazidime levels in newborns. This technique supports crucial pharmacokinetic studies for treating neonatal bacterial infections.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Neonatal Medicine
Background:
- Bacterial infections are a major cause of neonatal mortality.
- Ceftazidime is vital for treating neonatal infections, but its use is often off-label.
- Traditional blood sampling for pharmacokinetic studies in neonates is challenging.
Purpose of the Study:
- To develop and validate an ultra-performance liquid chromatography with a photodiode array (UPLC-PDA) method.
- To quantify ceftazidime in dried blood spots (DBS) from neonatal patients.
- To support pharmacokinetic studies for neonatal infections.
Main Methods:
- Utilized Capitainer® device for 10 µL DBS collection.
- Extracted ceftazidime using 70% methanol/water with acetaminophen as internal standard.
- Employed an Acquity UPLC HSS T3 column with a 3.2-minute run time and PDA detection at 259 nm.
Main Results:
- The UPLC-PDA method demonstrated high accuracy (90.1–104.8%) and precision (4.8–11.7% CV).
- The quantification range for ceftazidime was 0.5–200 µg/mL.
- No interfering peaks were observed, confirming method specificity.
Conclusions:
- A validated UPLC-PDA method for ceftazidime in neonatal DBS was successfully developed.
- This microsampling technique facilitates pharmacokinetic studies in neonates, aiding treatment optimization.
- The method is applicable to clinical samples from neonatal patients.
