Assessing Plasma Meropenem Concentrations: Validation of an HPLC Protocol
Student Research Paper
Purpose: The purpose of this study was to validate a laboratory protocol for comparing the assessing the concentration of meropenem in plasma samples.
Methods: To validate the laboratory component of a clinical protocol, plasma samples with meropenem and an internal standard of ceftazidime were analyzed using a Dionex® Ultimate 3000 HPLC and both an Acclaim® 120 C18 guard column (4.6 x 10 mm, 5 µm silica), and an Acclaim® 120 C18 analytical column (4.6 x 150 mm, 5 µm silica). Samples analyzed included quality control, calibration standards, and spiked unknowns blinded to the analysts. Each sample was prepared according to protocol and run at ambient temperature with a flow rate of 1 mL/min and an injection volume of 10 µL, in a mobile phase composed of 92% 0.1M mixed phosphate buffer (pH 6.8) and 8% acetonitrile with a stabilizer buffer of 3-[N-morpholino]propanesulfonic acid (MOPS).
Results: After analysis of a calibration curve and spiked samples, the correct concentration (± 10 µg/mL) was determined for nine out of ten plasma samples. However, there was significant variation in the data obtained by individual analysts.
Conclusions: The HPLC method is a valid analytical protocol for determining plasma concentration of meropenem. However, further laboratory training of the analysts is needed before analyzing patient samples.
Meropenem, HPLC, cystic fibrosis, pediatric, nausea
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