Analytical method development and validation for simultaneous estimation of meropenem and vaborbactam in bulk and pharmaceutical dosage form by RP-HPLC

Abstract

A simple, specific and accurate reverse phase high performance liquid chromatographic method was developed for the simultaneous determination Meropenemand Vaborbactaminpharmaceutical dosage form. The column used was KromosilC18(150mm x 4.6 mm, 5m)in isocratic mode, with mobile phase containing phosphatebufferandacetonitrile (45:55v/v). The buffer is prepared by adding accurately weighed 1.36gm of PotassiumdihyrogenOrtho phosphate in a 1000ml of Volumetric flask add about 900ml of milli-Q water added and degasto sonicate and finally make up the volume with water then pH adjusted to 5.0with dil. Orthophosphoric acid solution. The flow rate was 1.0ml/ min and effluents were monitored at 260nm. The retention times of Meropenemand Vaborbactamwere found to be 2.299 min and 3.102 min, respectively. The linearity for Meropenemand Vaborbactamwere in the range of 25-150μg/mland 25-150 μg/ml respectively. Regression equation of Meropenem is y = 4826.x + 2593, and y = 4887.x + 6194 of Vaborbactamrespectively. The proposed method was validated and successfully applied to the estimation of Meropenemand Vaborbactamin combined tablet dosage forms.

Fast and Sensitive Method for Simultaneous Quantification of Meropenem and Vaborbactam in Human Plasma Microsamples by Liquid Chromatography–Tandem Mass Spectrometry for Therapeutic Drug Monitoring

“…Single-charge positive ion mass transitions were selected for optimal sensitivity and specificity by analyzing the MS/MS fragmentation pattern spectra of analytes (data not shown) and by reviewing the literature [ 17 , 18 , 19 ]. Proton ion adducts of MRP and meropenem-d6 (MRP-d6) were detected by means of the following mass transitions: 384.2–141.0 m / z and 390.2–147.1 m / z and daughter ion intensity over retention time were used to generate multiple reaction monitoring (MRM) ion extraction chromatograms.…”

“…We are well aware that in LC-MS/MS methods isotopically labeled analogue should be used as the internal standard of the corresponding analyte, but this was unfeasible due to commercial unavailability at the time of the study. Consequently, we used AVI-C13 as IS, similar to what was previously done by other methods assessing MER/VBR plasma concentrations [ 17 , 18 , 19 ], with good results. Finally, the short stability of MRP, namely an issue very well-known in literature [ 17 , 20 ], was confirmed in the stability test conducted in our study, whereas VBR showed longer stability.…”

“…Consequently, we used AVI-C13 as IS, similar to what was previously done by other methods assessing MER/VBR plasma concentrations [ 17 , 18 , 19 ], with good results. Finally, the short stability of MRP, namely an issue very well-known in literature [ 17 , 20 ], was confirmed in the stability test conducted in our study, whereas VBR showed longer stability. Accordingly, to prevent degradation during transport, blood samples must be delivered to the lab without any delay and must be processed promptly or alternatively should be stored at −80 °C after blood centrifugation and plasma separation.…”

“…Several high-performance liquid chromatography (HPLC) or mass spectrometry (LC-MS/MS) methods have been developed for measuring MRP in biological fluids, either in plasma [ 8 , 9 , 10 , 11 ] or in serum [ 12 , 13 , 14 , 15 ] However, there are only a few available methods for measuring simultaneously MRP and VBR in biological fluids by means of HPLC [ 8 , 16 ] or of LC-MS/MS methods [ 17 , 18 , 19 ], and all of these are based on traditional blood sampling volume, namely 3–5 mL.…”