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An improved method for assaying phosphatidylcholine in mouse tissue.
Shigeo Murai1, Hiroko Saito, Rhuichi Shirato
1Department of Pharmacology, School of Dentistry, Iwate Medical University, Morioka, 020-8505, Japan.
Journal of Pharmacological and Toxicological Methods
|November 3, 2004
Summary
This study introduces a fast and accurate method for measuring phosphatidylcholine (PtdCh) in mouse tissues using HPLC-ECD. The new technique successfully detected changes in PtdCh levels in response to methoxamine, proving its utility for research.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Pharmacology
Background:
- Phosphatidylcholine (PtdCh) is a key phospholipid in cell membranes.
- Accurate measurement of PtdCh is crucial for understanding cellular processes and disease states.
- Existing methods for PtdCh quantification can be time-consuming and complex.
Purpose of the Study:
- To develop a rapid and precise method for measuring phosphatidylcholine (PtdCh) levels in mouse tissues.
- To validate the method's reliability and efficiency.
- To demonstrate the method's application by examining the effect of methoxamine on PtdCh levels in mouse artery and submandibular gland.
Main Methods:
- Development of a high-performance liquid chromatography (HPLC) method with electrochemical detection (ECD) and an immobilized enzyme column.
- Sample preparation involved direct injection of a hydrolyzed mixture of lipophilic choline metabolites.
- Utilized ethylhomocholine as an internal standard for choline assay.
Main Results:
- The developed HPLC-ECD method allowed for PtdCh assay within 5 minutes.
- High recovery (99%) and low coefficients of variation (0.6% and 1.3%) demonstrated method precision.
- Methoxamine administration significantly decreased PtdCh levels and increased free choline in mouse artery and submandibular gland.
Conclusions:
- The method enables direct injection of hydrolyzed tissue samples, simplifying PtdCh assay.
- The assay is highly reliable, rapid, and suitable for routine analysis of large sample numbers.
- This technique provides a valuable tool for PtdCh determination in various biological samples.