Simultaneous Determination of 12 β-Lactam Antibiotics in Human Plasma by High-Performance Liquid Chromatography with UV Detection: Application to Therapeutic Drug Monitoring

Abstract

A rapid and specific high-performance liquid chromatography method with UV detection (HPLC-UV) for the simultaneous determination of 12 beta-lactam antibiotics (amoxicillin, cefepime, cefotaxime, ceftazidime, ceftriaxone, cloxacillin, imipenem, meropenem, oxacillin, penicillin G, piperacillin, and ticarcillin) in small samples of human plasma is described. Extraction consisted of protein precipitation by acetonitrile. An Atlantis T3 analytical column with a linear gradient of acetonitrile and a pH 2 phosphoric acid solution was used for separation. Wavelength photodiode array detection was set either at 210 nm, 230 nm, or 298 nm according to the compound. This method is accurate and reproducible (coefficient of variation [CV] < 8%), allowing quantification of beta-lactam plasma levels from 5 to 250 g/ml without interference with other common drugs. This technique is easy to use in routine therapeutic drug monitoring of beta-lactam antibiotics.

Simultaneous determination of seven β-lactam antibiotics in human plasma for therapeutic drug monitoring and pharmacokinetic studies

“…For a comprehensive review of advances in the determination of -lactam antibiotics by liquid chromatography using different detector systems, the reader is referred to the paper by Lara et al [44]. In general, a number of HPLC methods with ultraviolet detection are described for -lactams including simultaneous determination of several -lactams [45][46][47][48][49].…”

“…The primary stocks were then diluted with water to give 5 mg/mL standard solutions for each antibiotic and were stored in 1 mL aliquots at -80 °C under which condition -lactams are known to be stable at least for few months [45,46,50,51]. These standard solutions were used to prepare fresh standards and QCs in plasma for every analytical run.…”

Sample Cleanup Using Solid-Phase Dispersive Extraction for Determination of Vancomycin in Serum

“…Some physicochemical methods such as liquid chromatography/ ultraviolet spectrometry (LC/UV), sensitive fluorometry (FL), and mass spectrometry (MS) were also used to correctly determine blood concentration. [11][12][13][14][15] However, a sample cleanup procedure (deproteinization extraction and solid-phase extraction (SPE)) is indispensable for eliminating the impurities from the sample matrix when performing such instrumental analysis.…”

The use of liquid chromatography-tandem mass spectrometry for therapeutic drug monitoring of antibiotics in cancer patients

“…Therefore, with the current limitations the use of immunoassays for antibiotics' TDM is not considered as a priority for further development and efforts have been directed to explore other alternatives such as the use of chromatographic techniques. Consequently, liquid-chromatography (LC) coupled to UV [28,[56][57][58][59], chemiluminescence (CL) [60], electrochemical (EC) [61], and mass spectrometry (MS) [62,63], have been widely/successfully used for the detection of β-lactams. Although as compared to mass spectrometry less sophisticated analysis is performed with the use of the other conventional detectors, the speed and sensitivity of the former one surpass those offered by the latter ones (UV, CL, EC).…”

“…Several studies published so far support the idea that using the conventional dosage regimens recommended for patients with minor infections is not adequate for critically ill patients with severe infections. Compelling evidence demonstrates that the dosing recommendation of β-lactam antibiotics is inadequate in critically ill patients [19] and that the poor clinical outcome observed to date is related to poor antibiotics exposure [13,18,19,[26][27][28].…”