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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 857; no. 1; pp. 32 - 39
• Main Authors: Xia, Su-Mei, Shen, Rong, Sun, Xue-Ying, Shen, Li-Li, Yang, Yi-Ming, Ke, Ying, Wang, Yun, Chen, Dong-Ying, Han, Xing-Mei
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 15.09.2007; Elsevier Science
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Animals; Anti-Inflammatory Agents, Non-Steroidal - analysis; Anti-Inflammatory Agents, Non-Steroidal - chemistry; Anti-Inflammatory Agents, Non-Steroidal - pharmacokinetics; Benzoates - analysis; Benzoates - chemistry; Benzoates - pharmacokinetics; Biological and medical sciences; Brain - metabolism; Brain Chemistry; Brain distribution; Bridged-Ring Compounds - analysis; Bridged-Ring Compounds - chemistry; Bridged-Ring Compounds - pharmacokinetics; Calibration; Chromatography, Liquid - methods; Chromatography, Liquid - standards; Drugs, Chinese Herbal - analysis; Drugs, Chinese Herbal - chemistry; Drugs, Chinese Herbal - pharmacokinetics; Fundamental and applied biological sciences. Psychology; General pharmacology; Glucosides - analysis; Glucosides - chemistry; Glucosides - pharmacokinetics; Injections, Subcutaneous; Iridoids - standards; LC–MS/MS; Male; Matrix effects; Medical sciences; Molecular Structure; Monoterpenes; Paeonia - chemistry; Paeoniflorin; Pharmacokinetics; Pharmacology. Drug treatments; Plant Roots - chemistry; Pyrans - standards; Rats; Rats, Sprague-Dawley; Reference Standards; Reproducibility of Results; Sensitivity and Specificity; Solid Phase Extraction; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry - methods; Tandem Mass Spectrometry - standards; Tissue Distribution; Matrix effects; Paeoniflorin; Brain distribution; Pharmacokinetics; LC–MS/MS; Validation; Terpenoid; Aldose; Rat; Rodentia; Glucoside; Central nervous system; Liquid chromatography; Matrix effect; Determination; Encephalon; LC-MS/MS; Vertebrata; Mammalia; Animal; Distribution; Development; Monoterpene; Glycoside; Quantitative analysis
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2007.06.022

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 × 2.1 mm i.d., 5 μm), using a mobile phase of methanol/water with 0.1% formic acid (50:50, v/v) at a flow-rate of 200–300 μL/min in 4 min. 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.