A simple HPLC method for the determination of S-adenosylmethionine and S-adenosylhomocysteine in rat tissues: the effect of vitamin B6 deficiency on these concentrations in rat liver.
A simple method using isocratic HPLC with ultraviolet detection was established for the simultaneous measurement of S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH) in rat tissues. The method provides rapid resolution of both compounds in a single run by direct injection of the perchloric acid extract of tissue, so that sampling procedures and analytical errors can be reduced in determining the SAM/SAH ratio, a measure of transmethylation reactions. The assay has a detection limit of 25 pmol and is linear in the range 50-200 pmol. This procedure was applied to determine the change of SAM/SAH ratio in liver caused by vitamin B6 deficiency. The marked reduction of the SAM/SAH ratios in vitamin B6-deficient rats were due to a drastic rise in SAH concentration with a concurrent striking decrease in SAM concentration.
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United Graduate School of Agricultural Science, Gifu University, Japan.
Nagao
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I
Hayakawa
T
T
Tsuge
H
H
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D008099:Liver / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012435:S-Adenosylhomocysteine / Q000378:metabolism*; D012436:S-Adenosylmethionine / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D026681:Vitamin B 6 Deficiency / Q000378:metabolism*
D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine