Simple determination of acetylcholine and choline within 4 min by HPLC-ECD and immobilized enzyme column in mice brain areas.
A rapid and simple method using a combination of high-performance liquid chromatography (HPLC) with electrochemical detection (ECD) and an immobilized enzyme column herein is described for simultaneous assay of acetylcholine (ACh) and choline (Ch) in discrete brain areas of the mouse. Perchloric acid extracts of small brain tissues were injected directly onto the HPLC system with no pre-cleanup procedure. The ACh and Ch levels were determined within 4 min for each chromatographic run. Recoveries were 100.0 +/- 4.8% for ACh and 96.1 +/- 1.4% (mean +/- S.D., n = 10) for Ch. Detection limits for both compounds were 0.25 pmol (signal-to-noise ratio of three). The present method has been applied to the measurement of Ch and ACh in several discrete brain areas of the mouse.
Murai
S
S
Department of Pharmacology, School of Dentistry, Iwate Medical University, Morioka, Japan.
Miyate
H
H
Saito
H
H
Nagahama
H
H
Masuda
Y
Y
Itoh
T
T
D000109:Acetylcholine / Q000032:analysis*; D000818:Animals; D001923:Brain Chemistry; D002794:Choline / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry; D004800:Enzymes, Immobilized; D008297:Male; D051379:Mice; D010472:Perchlorates; D013997:Time Factors
D004800:Enzymes, Immobilized; D010472:Perchlorates; D002794:Choline; D000109:Acetylcholine