A Vancomycin HPLC Assay for Use in Gut Microbiome Research

Area of Science:

• Microbiology
• Pharmacology
• Analytical Chemistry

Background:

• The human microbiome plays a critical role in health.
• Broad-spectrum antibiotics significantly perturb the gut microbiome.
• Standardized methods are needed to study microbiome pharmacodynamics.

Purpose of the Study:

• To validate a high-performance liquid chromatography (HPLC) method for quantitative fecal vancomycin analysis.
• To establish reproducible methodologies for vancomycin pharmacokinetic studies in the context of the human microbiome.

Main Methods:

• Validated a rapid HPLC-photodiode array detector method for fecal vancomycin quantification.
• Assessed reproducibility based on sample storage, homogeneity, and lyophilization.
• Determined vancomycin recovery (85-110%) and quantitative range (0.4-100 μg/mL).

Main Results:

• Achieved vancomycin elution in ~4.2 min with excellent recovery and a wide quantitative range.
• Demonstrated low intra-day and inter-day variability (RSD 0.4-5.4%).
• Confirmed consistent vancomycin concentrations across different stool aliquot weights and after lyophilization (RSD < 10%).

Conclusions:

• The validated HPLC method provides a reliable and reproducible approach for fecal vancomycin analysis.
• Established sample handling standards crucial for accurate vancomycin pharmacokinetic studies.
• This work supports research into the effects of antibiotics on the human microbiome.