Development and validation of a sensitive liquid chromatography-tandem mass spectrometry method for the determination of paeoniflorin in rat brain and its application to pharmacokinetic study.
A sensitive and specific method was developed and validated for the determination of paeoniflorin in rat brain with liquid chromatography-tandem mass spectrometry. Sample pretreatment involved protein precipitation following solid-phase extraction. Paeoniflorin and geniposide (internal standard) were separated isocratically on a Waters Symmetry C18 column (150 mm x 2.1 mm i.d., 5 microm), using a mobile phase of methanol/water with 0.1% formic acid (50:50, v/v) at a flow-rate of 200-300 microL/min in 4min. A Finngan LTQ tandem mass spectrometer equipped with electrospray ionization source was operated in the positive ion mode. Selective reaction monitoring was performed to quantify paeoniflorin and the internal standard at m/z transitions of 503-->381 and 411-->231, respectively. A good linearity was found in the range of 2-500 ng/mL (R(2)=0.9939). The intra- and inter-batch assay precisions (coefficient of variation, CV) at 5, 50 and 400 ng/mL (n=5) ranged from 6.3% to 9.7% and 1.2% to 7.2%, respectively, and the accuracies were from 95.9% to 101.6% and 99.4% to 102.9%, respectively. The mean recoveries of paeoniflorin were 81.2%, 80.9% and 82.3% at 5, 50 and 400 ng/mL (n=5), respectively, and the mean recovery of the internal standard was 76.7% with a concentration of 50 ng/mL (n=5). Stability studies showed that paeoniflorin was stable in different conditions. Finally, the method was successfully applied to the pharmacokinetic study of paeoniflorin in rat brain following a single subcutaneous administration (10 mg/kg) to rats.
Xia
Su-Mei
SM
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, PR China.
Shen
Rong
R
Sun
Xue-Ying
XY
Shen
Li-Li
LL
Yang
Yi-Ming
YM
Ke
Ying
Y
Wang
Yun
Y
Chen
Dong-Ying
DY
Han
Xing-Mei
XM
D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D001565:Benzoates / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D001952:Bridged-Ring Compounds / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods / Q000592:standards*; D004365:Drugs, Chinese Herbal / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D005960:Glucosides / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D007279:Injections, Subcutaneous; D039823:Iridoids / Q000592:standards; D008297:Male; D015394:Molecular Structure; D039821:Monoterpenes; D029596:Paeonia / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D011714:Pyrans / Q000592:standards; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods / Q000592:standards*; D014018:Tissue Distribution; D006027:Glycosides
D000894:Anti-Inflammatory Agents, Non-Steroidal; D001565:Benzoates; D001952:Bridged-Ring Compounds; D004365:Drugs, Chinese Herbal; D005960:Glucosides; D039823:Iridoids; D039821:Monoterpenes; D011714:Pyrans; C007835:geniposide; C015423:peoniflorin; D006027:Glycosides