Simultaneous determination of four active components in rat plasma by ultra-high performance liquid chromatography tandem-mass spectrometry/mass spectrometry and its application to a pharmacokinetic study after oral administration of Callicarpa nudiflora extract

Pharmacogn Mag. 2015 Jul-Sep;11(43):509-17. doi: 10.4103/0973-1296.160458.


Background: Callicarpa nudiflora has been commonly used as a Chinese folk medicine for resolving toxin, dispersing edema and hemostasis; however, its pharmacokinetic (PK) behavior remains unknown. In our present study, a simple and sensitive ultra-high performance liquid chromatography tandem mass spectrometry method was firstly developed on simultaneous determination and PK study of four active components (luteoloside, dracocephaloside, juncein and nudifloside) following the oral administration of C. nudiflora extract to investigate their PK profiles.

Materials and methods: Chromatographic separation was achieved on a Phenomenex(®) Kinetex C18 column (50 mm × 2.1 mm, 1.7 μm) with gradient elution using a mobile phase consisted of acetonitrile (A) and 0.05‰ formic acid in water (B). The quantitation was carried out by multiple reaction monitoring using electrospray ionization in the negative ion mode.

Results: Calibration curves offered satisfactory linearity, with correlation coefficients >0.99 for all compounds within the concentration range. The low limits of quantification were 1.03 ng/mL for luteoloside, 1.16 ng/mL for dracocephaloside, 0.82 ng/mL for juncein and 0.88 ng/mL for nudifloside, respectively. The intra- and inter-day precisions (relative standard deviation) were within 7.4% and the accuracies (relative error) ranged from -7.4% to 7.9%.

Conclusion: This method was successfully applied to the PK studies of luteoloside, dracocephaloside, juncein and nudifloside in rat plasma after oral administration of C. nudiflora extract, four analytes exhibited quick absorption with peak concentrations occurring at around 25 min and eliminated rapidly.

Keywords: Callicarpa nudiflora; determination; pharmacokinetics; rat plasma; ultra-high performance liquid chromatography tandem mass spectrometry.