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Fast analysis using pillar array columns: Quantification of branched-chain α-keto acids in human plasma samples
Takuya Fujiwara1, Takashi Funatsu1, Makoto Tsunoda1
1Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 1130033, Japan.
Journal of Pharmaceutical and Biomedical Analysis
|March 16, 2021
Summary
A new method rapidly quantifies branched-chain α-keto acids (BCKAs) in human plasma. This technique offers sensitive and precise analysis for BCKAs, aiding in clinical diagnosis of related diseases.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Clinical Diagnostics
Background:
- Branched-chain α-keto acids (BCKAs) are linked to serious diseases like cancer and diabetes.
- Accurate quantification of BCKAs is crucial for disease monitoring and diagnosis.
Purpose of the Study:
- To develop a rapid, sensitive, and precise analytical method for quantifying BCKAs.
- To apply the developed method for BCKA analysis in human plasma samples.
Main Methods:
- BCKAs were labeled with 1,2-diamino-4,5-methylenedioxybenzene (DMB).
- Separation was performed on octadecylsilane-treated pillar array columns using a MeOH/H2O mobile phase.
- Analysis time was reduced to 160 seconds for five DMB-labeled α-keto acids.
Main Results:
- Lower limits of quantification for DMB-α-keto acids ranged from 2-5 μM.
- Intra- and interday precisions were 2.9-6.6% and 5.2-10.7%, respectively.
- Plasma concentrations of KIV, KIC, and KMV were determined as 13.8, 24.2, and 15.2 μM.
Conclusions:
- The developed method provides rapid, sensitive, and precise BCKA quantification.
- This analytical approach is suitable for clinical diagnosis and disease management.
