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Updated: Aug 30, 2025
Multiplex Therapeutic Drug Monitoring by Isotope-dilution HPLC-MS/MS of Antibiotics in Critical Illnesses
Published on: August 30, 2018
Simple determination of urine cefazolin concentration in pediatric patients with urinary tract infections using
Kurumi Abe1,2, Kenji Momo2, Yoshifusa Abe3
1Department of Pharmacy, Showa University Koto Toyosu Hospital, Tokyo, Japan.
Insights
A new HPLC-UV method accurately measures cefazolin in pediatric urine, aiding antibiotic stewardship for urinary tract infections (UTIs) and combating multidrug resistance.
Area of Science:
- Analytical Chemistry
- Pharmacology
- Pediatrics
Background:
- Rising global concern over multidrug resistance necessitates effective antibiotic monitoring.
- Urinary tract infections (UTIs) in children require appropriate antibiotic therapy.
- First-generation cephem antibiotics like cefazolin are crucial for treating UTIs.
Purpose of the Study:
- To develop a simple, rapid, and inexpensive HPLC-UV method for quantifying urinary cefazolin concentrations.
- To validate the method for use in pediatric patients with UTIs.
- To support therapeutic drug monitoring and antibiotic stewardship.
Main Methods:
- High-performance liquid chromatography-ultraviolet (HPLC-UV) analysis at 254 nm.
- Urine samples diluted 10-fold with cephapirin as an internal standard.
- Linear calibration curve for cefazolin (31.25–500 μg/ml, r² > 0.999).
Main Results:
- The method demonstrated good linearity and precision (within- and between-day CVs: 1.2–19.2%).
- Cefazolin and internal standard retention times were 4.2 and 4.9 min, respectively.
- A pediatric UTI patient showed a urinary cefazolin concentration of 1,476.6 μg/ml.
Conclusions:
- The developed HPLC-UV method is suitable for therapeutic drug monitoring of cefazolin in pediatric UTIs.
- Accurate monitoring can ensure appropriate antibiotic use, potentially preventing antimicrobial resistance.
- This contributes to responsible antibiotic prescribing in pediatric healthcare.
Abstract:
Recently, global health concerns regarding increasing multidrug resistance have arisen. This study aimed to develop a simple, inexpensive and rapid high-performance liquid chromatography-ultraviolet (HPLC-UV) method for determining urinary concentrations of a first-generation cephem antibiotic in pediatric patients with urinary tract infections (UTIs). HPLC-UV was used to analyze urinary cefazolin concentrations at a detection wavelength of 254 nm. The assay used contained 10-fold diluted urine with an internal standard (cephapirin). The standard calibration curve for cefazolin was linear in the concentration range of 31.25-500 μg/ml (r2 > 0.999). The retention times of cefazolin and the internal standard were 4.2 and 4.9 min, respectively. The within- and between-day coefficients of variation were in the concentration ranges 1.2-15.2 and 5.5-19.2%, respectively. The urinary cefazolin concentration of a pediatric patient with a UTI was 1,476.6 μg/ml, which was over 700-fold higher than the minimum inhibitory concentration of cefazolin (≤2 μg/ml). The developed method is applicable to the confirmation of appropriate use for UTI treatment as therapeutic drug monitoring of cefazolin. Therefore, the findings of this study may contribute to the appropriate use of antibiotics to prevent antimicrobial resistance in pediatric patients with UTIs.
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