High-performance liquid chromatographic analysis of a new neuroprotective agent for ischemia-reperfusion damage, KR-31378.
A high-performance liquid chromatographic method was developed for the determination of a neuroprotective agent for ischemia-reperfusion damage, KR-31378, in human plasma and urine and in rat tissue homogenates. The method involved deproteinization of the the biological samples with 0.5 volumes of saturated Ba(OH)2, 0.5 volumes of 0.04 M ZnSO4 and 1 volume of acetonitrile. A 80-microl aliqout of the supernatant was injected onto a reversed-phase C18 column. The mobile phase, 50 mM triethylamine acetate : acetonitrile : tetrahydrofuran (65:30:5, v/v/v), was run at a flow rate of 1.0 ml/min. The column effluent was mornitored by a ultraviolet detector set at 310 nm. The retention time of KR-31378 was approximately 6.5 min. The detection limits of KR-31378 in human plasma and urine and rat tissue homogenates were 0.2, 0.5 and 0.5 microg/ml, respectively. The coefficients of variation (within-day and between-day) were below 13.6% for human plasma and urine and rat homogenates. No interferences from endogenous substances were found.
Lee
M H
MH
College of Pharmacy, Seoul National University, Shinlim-Dong, Kwanak-Gu, South Korea.
Kim
H J
HJ
Kim
S H
SH
Kim
S O
SO
Lee
D H
DH
Lim
H
H
Yoo
S E
SE
Lee
M G
MG
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D006146:Guanidines / Q000032:analysis* / Q000097:blood / Q000652:urine; D006801:Humans; D007202:Indicators and Reagents; D018696:Neuroprotective Agents / Q000032:analysis* / Q000097:blood / Q000652:urine; D011714:Pyrans / Q000032:analysis* / Q000097:blood / Q000652:urine; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D015427:Reperfusion Injury / Q000188:drug therapy*; D013056:Spectrophotometry, Ultraviolet; D014018:Tissue Distribution
D006146:Guanidines; D007202:Indicators and Reagents; C438686:N''-cyano-N-(6-amino-3,4-dihydro-3-hydroxy-2-methyl-2-dimethoxymethyl-2H-benzopyran-4-yl)-N'-benzylguanidine; D018696:Neuroprotective Agents; D011714:Pyrans