Simultaneous Liquid Chromatography–Mass Spectrometry Quantification of Cefixime and Clavulanic Acid in Human Plasma

Abstract

A simple and specific liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry (LC-APCI-MS) assay method has been developed and fully validated for the simultaneous quantification of cefixime (CX) and clavulanic acid (CA) in human plasma. Analytes and internal standard were extracted from human plasma by a solid phase extraction technique using a Sam prep (3 mL, 100 mg) extraction cartridge. The extracted samples were chromatographed on a reverse phase C18 column using a mixture of methanol : acetonitrile : 2 mM ammonium acetate (pH 3.5) (25 : 25 : 50, v/v/v) as the mobile phase at a flow rate of 0.8 mL/min. Quantification of the analytes were carried out using single quadrupole LC-APCI-MS through selected ion monitoring at m/z 452 and 198, respectively, for CX and CA. The assay was linear over the concentration range of 0.05-10.0 and 0.1-10.0 μg/mL, respectively, for CX and CA. The mean plasma extraction recoveries of the CX and CA were found to be 95.20-96.27% and 94.67-95.58%, respectively. The method was successfully applied for the determination of pharmacokinetics of CX and CA after oral administration of single dosage CX/CA (200/125 mg) pill to the humans (n = 12).

Quantification of 16 cephalosporins in the aquatic environment by liquid chromatography‐tandem mass spectrometry

“…The core structure of cephalosporins Environmental monitoring of antimicrobials usually requires the use of powerful analytical chemistry tools such as LC-MS/MS to achieve reasonable separation, identification, and quantification of both unknown and known pharmaceutical compounds. A scientific literature survey for cephalosporins estimation revealed their determination by LC-MS/MS in different matrices, such as biological fluids [17], human plasma [18][19][20], and environmental samples [21][22][23][24]. Table 1 represents an objective comparison of the reported LC-MS methods for estimating cephalosporins in different matrices [17][18][19][20][21][22].…”

An improved HPLC-DAD method for clavulanic acid quantification in fermentation broths of Streptomyces clavuligerus

“…The internal standard used is not structurally related to CA LC-MS [8] 0.1 (LOQ) Applied for the analysis of plasma obtained after oral administration of cefixime/CA combination (200/125 mg)…”

“…Bioassay, spectrophotometry and high performance liquid chromatography (HPLC) methods have been developed and are widely used for CA quantification [5][6][7][8][9][10]. Early developed HPLC methods for the detection of CA applied a pre-column derivatization step, as CA has a low detection wavelength in the UV region and is poorly retained on C-18 reverse phase columns [6,11,12].…”

“…Foroutan et al [13] and Hoizey et al [15] reported an HPLC-UV method for the simultaneous quantification of CA and amoxicillin in human plasma, with an internal standard and a direct deproteinization of plasma with methanol, respectively. Recently, methods with high selectivity and sensitivity, such as liquid chromatography-mass spectrometry (LC-MS), have been reported for CA determination in biological samples [8][9][10]16,17]. Dubala et al [8] described an LC-MS method based on solid phase extraction for the simultaneous determination of CA and cefixime in human plasma using chloramphenicol as internal standard.…”

“…Recently, methods with high selectivity and sensitivity, such as liquid chromatography-mass spectrometry (LC-MS), have been reported for CA determination in biological samples [8][9][10]16,17]. Dubala et al [8] described an LC-MS method based on solid phase extraction for the simultaneous determination of CA and cefixime in human plasma using chloramphenicol as internal standard. However, for all the above mentioned methods, that use either isocratic or gradient elution, CA is simultaneously quantified with another antibiotic, such as amoxicillin, in human samples after oral administration [8][9][10]15].…”