Related Experiment Videos
Determination of elastase activity by reversed-phase high-performance liquid chromatography.
T A Stein1, J R Cohen, C Mandell
1Department of Surgery, Long Island Jewish Medical Center, New Hyde Park, NY 11042.
Journal of Chromatography
|January 6, 1989
Summary
This study introduces a new method to measure elastase activity using high-performance liquid chromatography. This technique accurately quantifies elastolytic activity, potentially aiding in the diagnosis of certain diseases.
Area of Science:
- Biochemistry
- Analytical Chemistry
Background:
- Elastase is a key enzyme involved in elastin degradation.
- Accurate measurement of elastolytic activity is crucial for understanding physiological and pathological processes.
Purpose of the Study:
- To develop and validate a novel method for quantifying elastase activity.
- To assess the utility of this method for measuring elastolytic activity in diseased tissues.
Main Methods:
- High-performance liquid chromatography (HPLC) with on-line post-column derivatization was employed.
- Elastin hydrolysis products, specifically Val-Pro, were separated and quantified.
- Fluorescamine derivatization was used for sensitive detection of fluorescent products.
Main Results:
- A linear relationship was established between peak area and Val-Pro concentration (0.2-16 µg).
- The method demonstrated consistent elastase activity measurements over 6 to 24 hours.
- The measured elastase activity was 0.95 ± 0.02 µg/h.
Conclusions:
- The developed HPLC method provides a reliable and sensitive means to measure elastase activity.
- This technique shows promise for assessing elastolytic activity in clinical settings, particularly in diseased tissues.