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Comparative Study
. 2014 Jun 15:961:71-6.
doi: 10.1016/j.jchromb.2014.05.002. Epub 2014 May 14.

Determination of meropenem in bacterial media by LC-MS/MS

Liusheng Huang  1 , Janus Haagensen  2 , Davide Verotta  3 , Patricia Lizak  4 , Francesca Aweeka  4 , Katherine Yang  4
Affiliations
Comparative Study

Determination of meropenem in bacterial media by LC-MS/MS

Liusheng Huang et al. J Chromatogr B Analyt Technol Biomed Life Sci. .

Abstract

To support the development of a dynamic in vitro human pharmacokinetic/pharmacodynamic simulation model for biofilm-mediated infections and study stability of meropenem, an LC-MS/MS method for the determination of meropenem in Luria Bertani (LB) media was developed and validated in an API2000 LC-MS/MS system. A partial validation was also performed in M9 media. Sample aliquots of 100μL (or 25μL for M9 media) were mixed with the internal standard (IS) ceftazidime and filtered. The filtrate was directly injected onto a C8 column eluted with ammonium formate (10mM, pH 4) and acetonitrile (0.1% formic acid) in a gradient mode. ESI(+) and MRM with ion pair m/z 384→68 for meropenem and m/z 547→468 for the IS were used for quantification. The calibration curve concentration range was 50 to 25,000ng/mL. The recovery was over 98%. In LB media, significant signal suppression was observed throughout the time period of detection when compared with mobile phase solvents, but the matrix effect was compensated well with the IS. In M9 media, much less signal suppression was observed. The method is simple, fast, and reliable. Using the method, stability of meropenem in LB and M9 media were tested. No significant degradation was observed for at least 8h in both LB media (37°C) and M9 media (30°C), but more than 15% degradation was observed overnight (∼20h). The method was transferred to an API5000 LC-MS/MS system using meropenem-d6 as the IS.

Keywords: LB media; LC–MS/MS; M(9); Meropenem; Stability.

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Figures

Figure 1
Figure 1
Chemical structures of meropenem, meropenem-d6 and ceftazidime.
Figure 2
Figure 2
Chromatograms of meropenem at 50ng/mL (LLOQ) in LB and M9 media. The same ion pair m/z 384/68 was monitored. In M9 media, API5000 (lower panel) is about 100-fold more sensitive compared to API2000 (middle panel), counting on a half of injection volume and 5-fold additional sample dilution.
Figure 3
Figure 3
A representative concentration-time profile of meropenem in an in vitro PK/PD biofilm model.
See this image and copyright information in PMC

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