HPLC and LC-MS analysis of sinomenine and its application in pharmacokinetic studies in rats.
OBJECTIVE
To improve and validate analytical methods based on HPLC and liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) for the quantitative measurement of sinomenine in rat plasma and brain tissue.


METHODS
The separation of analytes and the internal standard (IS), chloramphenicol, was performed on an Agilent TC-C18 column (250×4.6 mm, 5 μm). Blood samples were measured with a Surveyor photodiode array (PDA) detector at a wavelength of 263 nm. The LCQ DECA XP(Plus) mass spectrometer was operated in the multiple reactions monitoring mode using positive electrospray ionization, and the transition from the precursor ion (m/z 279) to the product ion (m/z 224) for sinomenine was measured in brain tissue.


RESULTS
Measurements were linear over the concentration range of 0.1-100 μg/mL for sinomenine in plasma and over the range of 0.01-5.00 μg/g for sinomenine in brain tissue. The intra- and inter-day variabilities were less than 10% of the relative standard deviation (RSD), and the extraction and recovery of sinomenine was 72.48%-80.26% from plasma and 73.75%-80.26% from brain tissue. The limit of quantification (LOQ) was 0.1 μg/mL for plasma, and 0.01 μg/g for brain tissue. Identification of sinomenine was reproducible at 0.5, 5, and 50 μg/mL in the plasma and at 0.05, 0.50, and 2.00 μg/g in brain tissue. The concentration of sinomenine measured in brain tissue after a single ip dose had a neuroprotective effect on H₂O₂-induced injury in PC12 cells in vitro.


CONCLUSIONS
Our methods offered a sensitivity within a wide linear concentration range for sinomenine. These methods were successfully applied to evaluate sinomenine pharmacokinetics over time in rat brain tissue after a single ip dose of 30 mg/kg.
Long
Li-hong
LH
Department of Pharmacology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Wu
Peng-fei
PF
Chen
Xiang-long
XL
Zhang
Zui
Z
Chen
Yu
Y
Li
Yi-yong
YY
Jin
You
Y
Chen
Jian-guo
JG
Wang
Fang
F
D000818:Animals; D001921:Brain / Q000378:metabolism; D002138:Calibration; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006861:Hydrogen Peroxide / Q000633:toxicity; D008297:Male; D009019:Morphinans / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics* / Q000494:pharmacology; D018696:Neuroprotective Agents / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics* / Q000494:pharmacology; D016877:Oxidants / Q000633:toxicity; D016716:PC12 Cells; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D014018:Tissue Distribution
D009019:Morphinans; D018696:Neuroprotective Agents; D016877:Oxidants; C009271:sinomenine; D006861:Hydrogen Peroxide