Chiral gas chromatographic assay with flame ionization detection for amphetamine enantiomers in microsomal incubates.
A chiral assay for amphetamine enantiomers in rat liver microsomal incubates is based on derivatization with (S)-(-)-N-(trifluoroacetyl)-prolyl chloride (S-TFPC), capillary chromatographic separation of the diastereomeric amide derivatives, and detection by a flame ionization detector. The method is capable of detecting low levels of S- or R-amphetamine. The assay is linear from 5 to 250 micrograms/mL for each enantiomer, and the limit of detection is 0.5 microgram/mL. The analytical method affords the average recoveries of 77.53 +/- 5.22% for R-amphetamine and 74.47 +/- 3.08% for S-amphetamine. The method allows the study of the metabolic depletion of S- and R-amphetamine in rat liver microsomal incubates. The time-dependent concentration of amphetamine enantiomers in rat liver microsomes was determined, and the stereoselectivity of amphetamine phase I metabolism was observed.
Zeng
S
S
Department of Chemistry, Zhejiang University, People's Republic of China.
Zhang
L
L
Chen
Y Z
YZ
D000662:Amphetamines / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D002138:Calibration; D002849:Chromatography, Gas / Q000379:methods*; D005410:Flame Ionization; D008862:Microsomes, Liver / Q000737:chemistry*; D051381:Rats; D013237:Stereoisomerism
D000662:Amphetamines