A Vancomycin HPLC Assay for Use in Gut Microbiome Research

• DOI: 10.1128/spectrum.01688-21
• Publication Date: 2022-05-10T13:01:55Z
• Authors: Chenlin Hu; Nicholas D. Beyda; Kevin W. Garey
• Keywords: 0301 basic medicine; metagenomics; vancomycin; Reproducibility of Results; clinical trial; Microbiology; QR1-502; Anti-Bacterial Agents; Gastrointestinal Microbiome; 3. Good health; drug distribution; glycopeptide; 03 medical and health sciences; Vancomycin; Humans; HPLC; Chromatography, High Pressure Liquid; Research Article

The human microbiome project has revolutionized our understanding of the interaction between commensal microbes and health. By far, biggest perturbation involves use broad-spectrum antibiotics excreted in gut. Thus, pharmacodynamics changes relation to drug exposure pharmacokinetics is an emerging field. However, reproducibility studies are necessary develop A simple fast high-performance liquid chromatography-photodiode array detector (HPLC) method was validated for quantitative fecal vancomycin analysis. Reproducibility results were tested based on sample storage time, homogeneity antibiotic within stool, concentration consistency after lyophilization. HPLC enabled complete elution ~4.2 min reversed-phase C18 column under isocratic mode, with excellent recovery (85% 110%) over a 4-log, range (0.4-100 μg/mL). Relative standard derivations (RSD) intra-day inter-day ranged from 0.4% 5.4%. Using stool aliquots various weights consistently demonstrated similar concentrations (mean RSD: 6%; range: 2-16%). After correcting water concentrations, obtained lyophilization original samples (RSD less than 10%). These methodologies establish condition standards enable pharmacokinetic microbiome.