Development and Validation of an HPLC-ESI/MS/MS Method for the Determination of Amoxicillin, Its Major Metabolites, and Ampicillin Residues in Chicken Tissues

Abstract

A method for the simultaneous analysis of amoxicillin (AMO), amoxicillin metabolites, and ampicillin residues in edible chicken muscle, liver, and kidney samples via high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI/MS/MS) was developed and verified. The extraction and purification procedures involved the extraction of the sample using a liquid-liquid extraction method with acetonitrile to eliminate the proteins. The chicken tissue extract was then injected directly onto an HPLC column coupled to a mass spectrometer with an ESI(+) source. The HPLC-ESI/MS/MS method was validated according to specificity, sensitivity, linearity, matrix effects, precision, accuracy, decision limit, detection capability, and stability, as defined by the European Union and Food and Drug Administration. The linearity was desirable, and the determination coefficients (r2 values) ranged from 0.9968 and 0.9999. The limits of detection and limits of quantification were 0.10–2.20 μg/kg and 0.30–8.50 μg/kg, respectively. The decision limits were 57.71–61.25 μg/kg, and the detection capabilities were 65.41–72.50 μg/kg, and the recoveries of the four target analytes exceeded 75% at the limits of quantification and exceeded 83% at 25, 50, and 100 μg/kg (n = 6 at each level), confirming the reliability of this method for determining these analytes and providing a new detection technology. For real sample analysis, this experiment tested 30 chicken tissue samples, only one chicken muscle, liver, and kidney sample were contaminated with 5.20, 17.45, and 7.33 μg/kg of AMO values, respectively, while other target compounds were not detected in the 30 tested chicken tissue samples.

Novel Development of RP-HPLC Method to Quantify Amoxicillin, Omeprazole and Rifabutin in Combination

“…27 The relative standard deviation values of the peak areas of ACN, OPE and RFN corresponded to the precision of less than 2% relative standard deviation. 28 The proposed method was preciseness than methods of Shahrokhian et al 22 (2.8%) and Hemanth et al 25 14 Raju et al 15 Fatma, 16 Marcel et al 17 Chen et al 18 Ademar et al 19 Vital et al 20 Shahrokhian et al 22 Alamen et al 23 Hemanth et al 25 Singh and Srivastava, 26 and Sachin et al 23 Specificity was assured by ample separation of ACN, OPE and RFN peaks from each other and from additional other peaks originated during 0.1N HCl/0.1N NaOH induced hydrolysis, 30% peroxide induced oxidation and dry heat/sun light induced degradation conditions. 29 In the specificity test, the angles of purity for ACN, OPE and RFN peaks were observed to be decreased than for the purity thresholds, in samples of 0.1N HCl/0.1N NaOH induced hydrolysis, 30% peroxide induced oxidation and dry heat/sun light induced degradation conditions.…”

“…Quanmin and Zhanjun (2006, spectrophotometry), 14 Raju et al (2016, RP-HPLC), 15 Fatma (2016, Electrochemical), 16 Marcel et al (2018, HPLC), 17 Chen et al (2019, HPLC-MS/MS) 18 and Ademar et al (2020, Electrochemical) 19 reported methods to quantify ACN. Vital et al (2009, LC-MS), 20 Preeta et al (2010, HPTLC), 21 Shahrokhian et al (2015, Voltametry), 22 and Alamen et al (2018, spectrophotometry) 23 reported methods to quantify OPE.…”

“…None of the methods reported quantified ACN, OPE and RFN simultaneously. [14][15][16][17][18][19][20][21][22][23] The methods of Fatma, 16 Chen et al 18 Vital et al 20 and Hemanth et al 25 were not utilized for analysing drug in tablet dose type. A stability demonstrating HPLC methodology was developed for the first time to analyse ACN, OPE and RFN simultaneously.…”

“…Thus, OPE suppresses the release of the gastric acid. Few analytical methods were described for quantifying ACN, [14][15][16][17][18][19] OPE [20][21][22][23] and RFN [24][25][26][27] alone in pharmaceutical formulations and samples of biological nature. No analytical method, for the ACN, OPE and RFN combined assay, has been published to date.…”

Optimized and Validated Spectrophotometric Method for the Determination of Ampicillin in Pharmaceutical Formulations

“…The extensive literature survey revealed that several analytical techniques, likely as high-performance liquid chromatography (HPLC) [2,3], capillary electrophoresis (CE) [4,5], Fouriertransform infrared spectroscopy (FTIR) [6] were available for the determination in bulk and pharmaceutical formulations. It can be quantified with liquid chromatography-mass spectrophotometry (LCMS) [7] and ultra-performance liquid chromatography-mass spectrophotometry (UPLC-MS) [8] in biological fluids. Most of the techniques are involved with the complicated procedure, extraction with toxic solvents, and expensive instrumentation and pretreatment of the sample before analysis.…”