ABSTRACT
Objective: To establish the methods for determination of gelsemine (GM) and koumine (KM) in human plasma. Methods :GM and KM in human plasma were extracted by SPE tubes and determined by HPLC-UV. The chromatographic conditions were as following: The column was Shim-Pack C18 (250 mm × 4.6 mm, 5 μm); The mobile phase consisted of methanol, water, and dibutylamine (58∶42∶0.01); The flow rate was 1.0 mL/min; The injection volume was 50 μL; Column temperature was 30℃; Detective wavelength was set at 263 nm. Results: GM and KM were in good linearity in 0.05-20 μg/mL (n = 6, r = 0.999 4) and 0.05-20 μg/mL (n = 6, r = 0.999 5), respectively. The limits of detection for GM and KM were both 50 ng/mL (S/N ≥ 10), respectively. The average recoveries of extraction for GM and KM were 90.88% and 87.84%, respectively. The average recoveries of method for GM and KM were 98.65% and 96.31%, respectively. Average inter-day RSD values for GM and KM were 9.81% and 10.63% as well as 7.79% and 8.24% in intra-day RSD, respectively. Conclusion: The established method for the determination of GM and KM in human plasma by SPE is sensitive, simple, accurate, reliable, and suitable for pharmacokinetics and toxicity study on Gelsemii Elegantis Herba.
ABSTRACT
Objective: To observe the action of Salvia miltiorrhiza extracts (SmE2, liposoluble constituents) on the formation of physical dependence induced by morphine in mice and the effects of SmE2 on the production of cyclic adenosine monophosphate (cAMP) and nitric oxide (NO) in brain of morphine-dependent mice. Methods: Morphine-dependent mice model was established by sc injection of morphine at a gradual increasing dosage. The effect of SmE2 on the withdrawal symptoms induced by Naloxone in morphine-dependent mice was observed. cAMP and NO content in brain were determined through radioimmunoassay and nitrate reductase, respectively. Results: SmE2 (80 mg/kg) could remarkably reduce the withdrawal symptoms in morphine-dependent mice. The cAMP concentration in brain showed no obvious change in morphine-dependent mice treated with SmE2, but NO content in brain could significantly be reduced. Conclusion: SmE2 has effect on relieving the withdrawal symptoms in morphine-dependent mice with down-regulation of NO content in brain as possible mechanism.