An improved method for assaying phosphocholine and glycerophosphocholine in mouse tissue.
BACKGROUND
To measure the levels of phosphocholine (PCh) and glycerophosphocholine (GPCh) in the tissues and organs of mice, we developed a simple and rapid method using high-performance liquid chromatography (HPLC) with electrochemical detection (ECD) and an immobilized enzyme column.


METHODS
Under our modifications of the separation procedure of Klein et al. [Neurochem. Int. 2 (1993) 293], PCh and GPCh in the hydrophilic phase of the homogenate samples were selectively hydrolyzed into free choline by alkaline phosphatase and a 0.4-N perchloric acid solution, respectively, and the resulting hydrolyzed mixtures were directly injected into the HPLC system for analysis.


RESULTS
The present method permits PCh or GPCh assay within 5 min in one chromatographic run. Recoveries from tissue samples were 97% for PCh and 101% for GPCh. The percentages of the crossover reaction to the authentic PCh and GPCh were 0.4% and 3.8%, respectively. The within-run coefficients of variation for choline derived from PCh and GPCh in the tissue samples were 1.2% and 1.4%, respectively.


CONCLUSIONS
The method is effective and has been applied to the measurement of PCh and GPCh levels in several tissues of mice.
Murai
S
S
Department of Pharmacology, School of Dentistry, Iwate Medical University, Morioka 020-8505, Japan. smurai@iwate-med.ac.jp
Saito
H
H
Shirato
R
R
Kawaguchi
T
T
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005997:Glycerylphosphorylcholine / Q000032:analysis*; D008297:Male; D051379:Mice; D010767:Phosphorylcholine / Q000032:analysis*; D014018:Tissue Distribution
D010767:Phosphorylcholine; D005997:Glycerylphosphorylcholine