Development and Validation of Ecofriendly HPLC-MS Method for Quantitative Assay of Amoxicillin, Dicloxacillin, and Their Official Impurity in Pure and Dosage Forms

Abstract

Novel, accurate, selective, and rapid high-performance liquid chromatography mass spectrometry method was developed for simultaneous analysis of amoxicillin trihydrate, dicloxacillin sodium, and their official impurity 6-aminopenicillanic acid. The chromatographic separation was carried out by applying the mixture on a C18 column (3.5 µm ps, 100 mm × 4.6 mm id) using acetonitrile:water (65 : 35 by volume) as a mobile phase within only 4 min. The quantitative analysis was executed using single quadrupole mass spectrometer in which electrospray ionization, selected ion monitoring, and negative mode were operated. The retention times were 1.61, 2.54, and 3.50 mins for amoxicillin, 6-aminopenicillanic acid, and dicloxacillin, respectively. The method was validated in linear ranges of 2–28 µg mL−1, 2–35 µg mL−1, and 1–10 µg mL−1 for amoxicillin, dicloxacillin, and 6-aminopenicillanic acid, respectively. The results obtained from the suggested HPLC/MS were statistically compared with those obtained from the reported HPLC method, where no significant difference appeared respecting accuracy and precision. According to the analytical eco-scale assessment method, the proposed method was proved to be greener than the reported one, where the analysis time and the amount of the wasted effluent decreased.

A Validated HPLC Multichannel DAD Method for the Simultaneous Determination of Amoxicillin and Doxycycline in Pharmaceutical Formulations and Wastewater Samples

“…The LOQs of the optimized method were low enough to perform the quantitative determination of Amox and Dox in a variety of matrices. Our study's LOD and LOQ values are much lower than those of 0.436 and 1.32 µg/mL [20], 2.14 and 7.15 µg/mL [54], 1.579 and µg/mL, 0.1 and 0.3 mg/mL [41], and 4.785 µg/mL [17] reported for the quantitative determination of Amox, and 1.08 and 3.27 µg/mL [55], and 16 and 54 µg/L [6] stated for Dox in pharmaceutical formulations and wastewater samples using the HPLC method.…”

“…No antibiotics (Amox and Dox) were detected in the wastewater collected from the sewage system of the two stations from Cluj-Napoca. Lower recovery (97.0 ± 1.6%) for the determination of Amox in spiked wastewater at 10 mg/L was found by Unutkan et al [6], while Almaki et al reported 100.99 ± 1.632% for the assay of Amox in capsules [20].…”

“…Amox has better absorption from the intestinal tract, higher capacity for achieving effective concentrations at the receptors, and an enhanced ability to penetrate Gram-negative microorganisms' cellular wall compared to other β-lactam antibiotics [7]. [8,9], voltammetry [10,11], fluorimetry [12,13], microbiological methods [14], capillary electrophoresis [15,16], and high-performance liquid chromatography (HPLC) [2,6,[17][18][19][20][21][22].…”

“…HPLC is a valuable tool in the quantification of antibiotics in various types of samples due to its simplicity, speed, low cost, and applicability to various analyte types. Moreover, HPLC has been renowned as a robust separation technique with high selectivity [20]. Unfortunately, several HPLC methods validated for the simultaneous determination of different antibiotics in the same sample suffer from several drawbacks, such as the need for extensive runtime or large sample volumes.…”

“…These characteristics make the designed method particularly appropriate for performing several daily measurements. Moreover, this method is applicable to small sample The literature depicts various analytical approaches for the quantitative determination of Amox in commercial drugs and wastewater samples, namely spectrophotometry [8,9], voltammetry [10,11], fluorimetry [12,13], microbiological methods [14], capillary electrophoresis [15,16], and high-performance liquid chromatography (HPLC) [2,6,[17][18][19][20][21][22].…”