Development of a validated HPLC method for the determination of four penicillin antibiotics in pharmaceuticals and human biological fluids

Abstract

A quantitative method for the determination of four penicillin antibiotics, amoxicillin (AMO), oxacillin (OXA), cloxacillin (CLO), and dicloxacillin (DICLO), has been developed. Separation was achieved on an Inertsil ODS-3 (250 x 4 mm, 5 microm) column after selective extraction of penicillin drugs from biological matrices by means of SPE. Gradient elution with a mobile phase consisting of 0.1% TFA (pH 1) and ACN, and PDA detection with monitoring at 240 nm was applied. Salicylic acid (5 ng/microL) was used as the internal standard. RP-8 Adsorbex Merck cartridges provided high absolute recoveries (98-101%). The developed method was fully validated in terms of selectivity, linearity, accuracy, precision, stability, and sensitivity. Repeatability (n = 8) and between-day precision (n = 8) revealed RSD <10%. Recoveries from biological samples ranged from 91 to 103%. The detection limits were estimated as 3.3 ng for AMO, OXA, and CLO, and 6.6 for DICLO in blood plasma. LOD in whole blood and urine was 6.6 ng. Injection volume was 20 microL. The method was applied to commercially available AMO containing pharmaceuticals and spiked biological matrices. The method was also applied to biological samples after AMO oral administration, where the drug was successfully identified and quantified.

Quantification of seven β-lactam antibiotics and two β-lactamase inhibitors in human plasma using a validated UPLC-MS/MS method

“…The major advantage of this method is its speed of analysis (5.5 min/sample), which is considerably faster than other methods for multiple analytes previously reported [15][16][17][18][19][20][21][22][23]. This allows high sample throughput and enables fast reporting of the results if this method were to be used for therapeutic drug monitoring.…”

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

“…There are several proposed methods for assaying beta-lactams in biological fluids, either for a particular compound (5,11,24,30) or for a group of compounds (9,10,13,17,18,21,29,33,34). Many other methods were developed for food industry and agriculture applications (1).…”

Identification of Penicillinase-Resistant Penicillins in Human Urine by RPLC Method

“…The basic structure of penicillin is a thiazolidine ring attached to a β-lactam ring. Five members of this class of antibiotics are determined in the present study: Oxacillin, methicillin, nafcillin, dicloxacillin and cloxacillin (Figure 1) [1][2].…”