Simultaneous determination of liensinine, isoliensinine and neferine from seed embryo of Nelumbo nucifera Gaertn. in rat plasma by a rapid HPLC method and its application to a pharmacokinetic study

Abstract

A rapid and specific HPLC method has been developed and validated for the simultaneous determination of liensinine (CAS 2586-96-1), isoliensinine (CAS 6817-41-0) and neferine (CAS 2292-16-2) in rat plasma. The sample was prepared by a liquid-liquid extraction with diethyl ether and the recovery was above 80 % from the plasma for the three compounds. Chromatographic separation was achieved with a Hypersil BDS C₁₈ column (4.0 mm × 250 mm, particle size 5 μm). A mobile phase consisting of methanol : 0.2 M KH₂P0₄:0.2 M NaOH : Methylamine (71:17:12:0.002, v/v/v/v, pH 9.2-9.3) was slowly delivered at 0.8 ml/min in isocratic mode with a detection wave-length of 282 nm. The linearity of calibration curves were good (r > 0.999) in the concentration range of 0.031 -2.00 μg/ml. The lower limit of quantification can reach 0.03 μg/ml for the three compounds. The intra-day and inter-day variations estimated with QC samples were less than 8 % for the three tested concentration levels. This developed method was applied in the plasma pharmacokinetic study of total bisbenzylisoquinoline alkaloids (TAL) of the lotus flower (Lian Zi Xin) following a single oral and intravenous administration of TAL in rats.

Key words

HPLC - Isoliensinine - Liensinine - Neferine - Pharmacokinetics

• References

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