High-performance liquid chromatographic analyser for guanidino compounds using benzoin as a fluorogenic reagent

Area of Science:

• Biochemistry
• Analytical Chemistry
• Clinical Diagnostics

Background:

• Biogenic guanidino compounds play crucial roles in human physiology.
• Accurate quantification of these compounds is essential for diagnosing various metabolic disorders.
• Existing analytical methods can be time-consuming or lack sensitivity.

Purpose of the Study:

• To develop an automated analytical system for quantifying biogenic guanidino compounds.
• To achieve rapid and sensitive separation and detection of fourteen key guanidino compounds.
• To establish a reliable method for analyzing these compounds in human physiological fluids.

Main Methods:

• Utilized high-performance liquid chromatography (HPLC) with a cation-exchange column.
• Employed a stepwise gradient elution system with varying pH and ionic strength.
• Automated derivatization to fluorescent products using benzoin for detection.

Main Results:

• Successfully separated fourteen guanidino compounds within 35 minutes.
• Achieved low limits of detection: 5-50 pmol for monosubstituted compounds, 1 nmol for creatine, and 20 nmol for creatinine.
• Demonstrated a simple, rapid, and sensitive analytical procedure.

Conclusions:

• The developed automated HPLC system provides efficient quantification of biogenic guanidino compounds.
• The method's sensitivity and speed make it suitable for clinical and research applications.
• This analyzer offers a valuable tool for studying guanidino compound metabolism and related diseases.