Determination of temazepam and temazepam glucuronide by reversed-phase high-performance liquid chromatography.
A rapid and sensitive method for extracting temazepam from human serum and urine is presented. Free temazepam is extracted from plasma and urine samples using n-butyl chloride with nitrazepam as the internal standard. Temazepam glucuronide is analyzed as free temazepam after incubating extracts with beta-glucuronidase. Separation is achieved using a C8 reversed-phase column with a methanol-water-phosphate buffer mobile phase. An ultraviolet detector operated at 230 nm is used and a linear response is observed from 20 ng/ml to 10 micrograms/ml. The limit of detection is 15.5 ng/ml and the limit of quantitation is 46.5 ng/ml. Coefficients of variation are less than 10% for concentrations greater than 50 ng/ml. Application of the methodology is demonstrated in a pharmacokinetic study using eight healthy male subjects.
Kunsman
G W
GW
Department of Pharmacology, Louisiana State University Medical Center, Shreveport 71130-3932.
Manno
J E
JE
Przekop
M A
MA
Manno
B R
BR
Llorens
K A
KA
Kunsman
C M
CM
D002851:Chromatography, High Pressure Liquid; D006801:Humans; D013056:Spectrophotometry, Ultraviolet; D013693:Temazepam / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism* / Q000652:urine
C072757:temazepam glucuronide; D013693:Temazepam