Simple and reliable extraction and a validated high performance liquid chromatographic assay for quantification of amoxicillin from plasma.

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dc.contributor.author Purohit, Trusha en
dc.contributor.author Wu, Zimei en
dc.contributor.author Hanning, Sara en
dc.date.accessioned 2020-02-13T03:41:06Z en
dc.date.issued 2020-01 en
dc.identifier.issn 0021-9673 en
dc.identifier.uri http://hdl.handle.net/2292/49999 en
dc.description.abstract This study presents the development of an efficient extraction protocol for amoxicillin from plasma with improved solubility and stability using pH control. Solubility and stability of amoxicillin in commonly used extraction solvents were determined using a newly developed stability-indicating high-performance liquid chromatography (HPLC) method. Following this, protein precipitation (PP) mediated sample purification protocol was developed and validated along with the HPLC method for the extracted amoxicillin from rabbit plasma. The protocol was applied in a pharmacokinetic study in rabbits. A five-fold increase in solubility and two-fold increase in stability of amoxicillin was found by addition of acetate buffer (0.1 M, pH 5.0) in acetonitrile. PP mediated extraction protocol containing acetate buffer-acetonitrile (1:18 v/v) resulted in an extraction recovery of >80% for all the samples. The HPLC assay following extraction was found linear (R2   >0.9999) over the range of 0.2-20 µg/mL with a lower limit of quantification of 0.2 µg/mL. The accuracy of the quality control samples was found between 97-115% and the relative standard deviation (RSD) was found to be below 6% for all samples. The samples were stable in the mobile phase (pH 5.0) for 72 h post-extraction. Amoxicillin-spiked plasma samples were found stable for up to three freeze-and-thaw cycles but, nearly 50% samples had degraded following storage for two months at -20 °C. Pharmacokinetic analysis indicated a half-life of amoxicillin of nearly 1 h following intravenous injection in rabbits, which is similar to that in humans. Thus, a simple and repeatable, extraction protocol was developed using pH control for quantification of amoxicillin from plasma based on its physicochemical properties. en
dc.format.medium Print-Electronic en
dc.language eng en
dc.relation.ispartofseries Journal of chromatography. A en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.subject Animals en
dc.subject Rabbits en
dc.subject Humans en
dc.subject Amoxicillin en
dc.subject Solvents en
dc.subject Chromatography, High Pressure Liquid en
dc.subject Reproducibility of Results en
dc.subject Kinetics en
dc.subject Solubility en
dc.title Simple and reliable extraction and a validated high performance liquid chromatographic assay for quantification of amoxicillin from plasma. en
dc.type Journal Article en
dc.identifier.doi 10.1016/j.chroma.2019.460611 en
pubs.begin-page 460611 en
pubs.volume 1611 en
dc.rights.holder Copyright: The author en
pubs.publication-status Published en
dc.rights.accessrights http://purl.org/eprint/accessRights/RestrictedAccess en
pubs.subtype Validation Study en
pubs.subtype Journal Article en
pubs.elements-id 784993 en
pubs.org-id Medical and Health Sciences en
pubs.org-id Pharmacy en
dc.identifier.eissn 1873-3778 en
pubs.record-created-at-source-date 2019-10-20 en
pubs.dimensions-id 31627968 en


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