Determination of gelsemine and koumine in human plasma by HPLC-UV / 中草药

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.