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Assays for angiotensin converting enzyme inhibitory activity.

B W Chang¹, R L Chen, I J Huang

¹Department of Healthcare Administration, Hungkuang Institute of Technology, Taichung, Taiwan, Republic of China

Analytical Biochemistry

|March 23, 2001

Summary

New methods quantify histidyl-leucine and hippurate. A sensitive colorimetric assay and a rapid capillary electrophoresis technique offer efficient analysis for biochemical research and enzyme activity measurement.

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Area of Science:

Analytical Chemistry
Biochemistry

Background:

Accurate quantification of specific biomolecules is crucial for biochemical research.
Histidyl-leucine and hippurate are relevant metabolites with implications in various biological processes.
Existing methods for their quantification may lack sensitivity or require extensive sample preparation.

Purpose of the Study:

To develop and validate a colorimetric method for quantifying histidyl-leucine.
To develop and validate a capillary electrophoresis (CE) method for quantifying hippurate.
To propose protocols for measuring angiotensin converting enzyme (ACE) inhibitory activity using these methods.

Main Methods:

A colorimetric assay utilizing o-phthalaldehyde for selective chromogenic reaction with histidyl-leucine.

Related Experiment Videos

Capillary electrophoresis with in-capillary microextraction for hippurate quantification.

Application of developed methods to assess ACE inhibitory activity.

Main Results:

The colorimetric method demonstrated high sensitivity (extinction coefficient = 7.5 mM(-1) cm(-1)) and reproducibility (CV = 1.7%).
The CE method allowed rapid ( < 4 min) and pretreatment-free analysis of hippurate, even in complex samples.
The methods were successfully applied to measure ACE inhibitory activity.

Conclusions:

Both developed methods provide sensitive and efficient means for quantifying histidyl-leucine and hippurate.
The CE method is particularly advantageous for complex samples and high-throughput analysis.
These methods offer valuable tools for biochemical analysis and drug discovery, specifically in evaluating ACE inhibitors.