Quantitative analysis of intracellular glutathione levels in murine bone marrow cells by using reverse-phase high-performance liquid chromatography.
Intracellular reduced glutathione (GSH) and oxidized glutathione (GSSG) amounts in murine bone marrow cells were determined by using a simple and rapid quantitative analysis that used reverse-phase high-performance liquid chromatography. Freshly prepared murine bone marrow cells lysed with water were applied to C18 double columns, were eluted with 0.1% trifluoroacetic acid-H2O at 35EC and were detected by ultraviolet absorption (220 nm). GSH and GSSG peaks with retention times of 9.2 and 27.5 min, respectively, were clear and were free of interference from the peaks of contaminated molecules in the cells. The results of this study indicate that our simple method to determine intracellular GSH and GSSG amounts can be used for a small number of cells (>3 x 10(6) cells).
Murakami
Miho
M
Department of Pharmaceutical Administration, Hokkaido College of Pharmacy, 7-1, Katsuraoka-cho, Otaru, 047-0264, Japan. miho-c-m@hokuyakudai.ac.jp
Kashiwakura
Ikuo
I
Hayase
Yukitoshi
Y
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D002138:Calibration; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D003114:Colony-Forming Units Assay; D005978:Glutathione / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000502:physiology; D051379:Mice; D010084:Oxidation-Reduction; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet
D019009:Proto-Oncogene Proteins c-kit; D005978:Glutathione