Determination of Huperzine A in rat plasma by high-performance liquid chromatography with a fluorescence detector.
A simple reversed-phase high-performance liquid chromatography (HPLC)-fluorescence method for the determination of Huperzine A in rat plasma was developed and validated. Separation was achieved on Kromasil C(8) column (5 microm, 150 mm x 4.6mm i.d.). The mobile phase, methanol-water-triethanol amine (45:55:0.05, v/v/v), was delivered at a flow rate of 1.0 ml/min. The eluent was monitored by a fluorescence detector with excitation wavelength at 310 nm and emission wavelength at 370 nm. No interfering peaks were observed in rat blank plasma. The relationship between Huperzine A concentration and peak-area ratio of Huperzine A to the IS was linear over the range of 2.5-250 ng/ml. The intra- and inter-day coefficients of variation were <or=10.1% and <or=14.1%, respectively. The method and extraction recovery of Huperzine A were 101.9-108.9% and 73.5-84.6%, respectively. Huperzine A was found to be stable for at least 5h at RT and 1 week at -20 degrees C. The method was applied to a pharmacokinetic study of Huperzine A in rats following intranasal administration.
Yue
Peng
P
Division of Pharmaceutics, Shanghai Institute of Pharmaceutical Industry, ZhongShanBeiYi Road 1111, Shanghai 200437, China.
Tao
Tao
T
Zhao
Yan
Y
Ren
Jinfeng
J
Chai
Xuyu
X
D000042:Absorption; D000281:Administration, Intranasal; D000470:Alkaloids; D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004355:Drug Stability; D004983:Ethanolamines / Q000737:chemistry; D005615:Freezing; D006207:Half-Life; D008657:Metabolic Clearance Rate; D000432:Methanol / Q000737:chemistry; D015394:Molecular Structure; D018696:Neuroprotective Agents / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012717:Sesquiterpenes / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors; D014867:Water / Q000737:chemistry
D000470:Alkaloids; D004983:Ethanolamines; D018696:Neuroprotective Agents; D012717:Sesquiterpenes; C050426:huperzine A; D014867:Water; C009546:triethanolamine; D000432:Methanol