Development of a high performance liquid chromatography method for quantification of PAC-1 in rat plasma.
A sensitive and specific high performance liquid chromatography method with UV detection was developed and validated for the determination of PAC-1 in rat plasma. After extraction with ethyl acetate, the chromatographic separation was carried out on a Diamonsil C(18) column (150mmx4.6mm i.d., 5microm particle size, Zhonghuida) protected by a ODS guard column (10mmx4.6mm i.d., 5microm particle size), using acetonitrile-methanol-phosphate buffer (pH 3.0, 30mM) (31:3:66, v/v/v) as mobile phase at a flow rate of 1.0mL/min, and wavelength of the UV detector was set at 281nm. No interference from any endogenous substances was observed during the elution of PAC-1 and internal standard (IS, indapamide). The calibration curve was linear over the range of 0.05-20microg/mL (r>0.99). The lower limit of quantification was evaluated to be 50ng/mL. The method was successfully applied to the pharmacokinetic study of PAC-1 after intravenous and oral administration in rats, respectively.
Fang
Li-Na
LN
Shenyang Pharmaceutical University, Shenyang 110016, People's Republic of China.
Chen
Xiao-Hui
XH
Song
Zhen
Z
Wang
Gang
G
Zhao
Xu
X
Ren
Lei
L
Gong
Ping
P
Bi
Kai-Shun
KS
D000818:Animals; D016588:Anticarcinogenic Agents / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D019540:Area Under Curve; D001682:Biological Availability; D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D004353:Drug Evaluation, Preclinical; D004355:Drug Stability; D004356:Drug Storage; D005215:Fasting; D005615:Freezing; D006207:Half-Life; D006835:Hydrazones / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008297:Male; D015394:Molecular Structure; D010316:Particle Size; D010710:Phosphates / Q000737:chemistry; D010879:Piperazines / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D011897:Random Allocation; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012997:Solvents / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
C514579:(4-benzylpiperazin-1-yl)acetic acid (3-allyl-2-hydroxybenzylidene)hydrazine; D016588:Anticarcinogenic Agents; D002021:Buffers; D006835:Hydrazones; D010710:Phosphates; D010879:Piperazines; D012997:Solvents