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Determination of urinary pyrraline by solid-phase extraction and high performance liquid chromatography
K Yoshihara1, R Kiyonami, Y Shimizu
1Laboratory of Public Health, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Japan.
Biological & Pharmaceutical Bulletin
|August 21, 2001
Summary
This study introduces a new method to measure pyrraline, an advanced glycation end product, in urine. The developed technique is simple, rapid, and accurate for assessing pyrraline levels in healthy individuals.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Clinical Chemistry
Background:
- Advanced glycation end products (AGEs) like pyrraline form in vivo under non-enzymatic, non-oxidative conditions.
- Pyrraline is a biomarker of glycation stress, but its accurate measurement in biological samples is challenging.
Purpose of the Study:
- To develop and validate a novel, efficient method for determining urinary pyrraline levels.
- To establish a baseline excretion level of pyrraline in healthy adults.
Main Methods:
- A new method utilizing solid-phase extraction (SPE) with an Oasis HLB cartridge for urine sample pretreatment.
- High-performance liquid chromatography (HPLC) for the quantitative determination of pyrraline post-SPE.
- Urine samples from healthy individuals (n=30, ages 20-77) were analyzed without prior hydrolysis.
Main Results:
- The SPE method effectively removed interfering substances, enabling clear detection of the pyrraline peak.
- High recovery rate of pyrraline (97.2+/-3.3%) was achieved during the SPE process.
- The mean urinary pyrraline excretion level in healthy controls was determined to be 1.42+/-0.65 micromol/mmol creatinine.
- Daily variations in pyrraline excretion levels were found to be insignificant in the study population.
Conclusions:
- The developed SPE-HPLC method is a simple, rapid, and accurate approach for quantifying urinary pyrraline.
- This method facilitates reliable assessment of pyrraline as a biomarker in clinical and research settings.
- The established baseline levels provide a reference for future studies on pyrraline-related health conditions.