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Assay of metronidazole by HPLC compared with microbial method
Scandinavian Journal of Gastroenterology. Supplement
|January 1, 1984
Summary
High-performance liquid chromatography (HPLC) offers superior sensitivity and precision for quantifying metronidazole and its active metabolite in serum compared to traditional microbiological assays. This advanced method accurately measures total drug and metabolite levels, crucial for therapeutic drug monitoring.
Area of Science:
- Analytical Chemistry
- Pharmacokinetics
- Microbiology
Background:
- Accurate quantification of metronidazole and its metabolites in biological fluids is essential for pharmacokinetic studies and therapeutic drug monitoring.
- Existing microbiological assays may not differentiate between active metabolites or account for protein binding.
Purpose of the Study:
- To compare a novel high-performance liquid chromatography (HPLC) method with a standard microbiological assay for metronidazole and its 2-hydroxymethyl metabolite in human serum.
- To evaluate the sensitivity, speed, and precision of the HPLC method.
Main Methods:
- Development and application of an HPLC method using a mu-Bondapak C18 column and UV detection for separating metronidazole and its metabolites.
- Utilized tinidazole as an internal standard for quantification by peak height ratios.
- Comparison with an agar well diffusion microbiological assay using Clostridium perfringens.
Main Results:
- The HPLC method demonstrated superior sensitivity (0-1 microgram/ml serum), speed, and precision.
- HPLC effectively separated metronidazole from its 2-hydroxymethyl and 1-carboxymethyl metabolites.
- The microbiological method determined total antibacterial activity, including free moieties, while HPLC quantified specific compounds.
Conclusions:
- The developed HPLC method is a highly sensitive, precise, and rapid technique for the simultaneous assay of metronidazole and its 2-hydroxymethyl metabolite in serum.
- HPLC provides a more detailed pharmacokinetic profile by distinguishing between the parent drug and its active metabolites.
- The findings support the use of HPLC for improved therapeutic drug monitoring of metronidazole therapy.