HPLC and LC-MS analysis of sinomenine and its application in pharmacokinetic studies in rats

Abstract

Aim: 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 H 2 O 2 -induced injury in PC12 cells in vitro. Conclusion: 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.

Sinomenine inhibits microglial activation by Aβ and confers neuroprotection

“…It has been shown that intraperitoneally injected or orally administered sinomenine can reach the brain [39,40] suggesting that it can cross blood-brain barrier. In addition, intraperitoneally injected sinomenine confers protection in ischemic brain injury [41].…”

Sinomenium acutum: A review of chemistry, pharmacology, pharmacokinetics, and clinical use

“…The experimental studies using beagle dog models demonstrate that the absolute bioavailability of sinomenine is low, and the elimination of sinomenine tablet is fast . After the quantitative measurement of sinomenine, Long et al (2010) found that the numerical values are 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 extraction and recovery of sinomenine from plasma and brain tissue are 72.…”

Determination of sinomenine sustained-release capsules in healthy Chinese volunteers by liquid chromatography–tandem mass spectrometry

“…Several liquid-liquid extractions or protein precipitation procedures [14][15][16][17][18][19][20][21][22] were exploited for plasma sample preparation. Acetonitrile, methanol and 7% perchloric acid were chosen and tested for protein precipitation because of high throughput characteristics and the suitable sensitivities obtained.…”

“…Currently, the primary focus of quantification of sinomenine hydrochloride in biological samples is based on the high-performance liquid chromatography with ultraviolet detector (LC-UV) [14][15][16][17][18][19][20], which required either a complicated extraction or long analysis time (>10 min). Furthermore, the lower limits of quantitation (LLOQ) of these methods in a range of 6-320 ng/mL were not suitable for sinomenine pharmacokinetic study.…”

“…Furthermore, the lower limits of quantitation (LLOQ) of these methods in a range of 6-320 ng/mL were not suitable for sinomenine pharmacokinetic study. Some more sensitive methods using LC/MS have been reported to detect this drug in rat's brain [20] or skin [21]. Only one study using ion trap mass spectrometry has been published for the determination of sinomenine in human plasma with an LLOQ of 0.5 ng/mL [22], but the analyses were performed only with spiked plasma samples so pharmacokinetic parameters were not available.…”