Abstract:

Objective: To develop a sensitive and selective high performance liquid chromatography (HPLC) method
for identification and quantification of alkylamide from Sichuan pepper (Zanthoxylum bungeanum) in various tissues
of rats administered with it. Methods: Chromatographic separation was performed on an intersil ODS-SP C18 column
(250 mm × 4.6 mm, 5 μm) using a mobile phase composed of a mixture of methanol and water with a ratio of
70:30 (V/V) at a flow rate of 0.5 mL/min. The detection wavelength was 254 nm. Results: Good linearity was obtained
for alkylamide in heart, liver, spleen, lung, kidney, brain, stomach, small intestine, fat, muscle, skin, testicular and
prostate over the ranges of 0.1–10 (r = 0.998 6), 0.1–50 (r = 0.999 9), 0.1–10 (r = 0.999 7), 0.1–10 (r = 0.998 9),
0.1–20 (r = 0.999 3), 0.1–10 (r = 0.999 5), 0.1–100 (r = 0.999 9), 0.1–50 (r = 0.998 2), 0.1–20 (r = 0.995 6), 0.1–10
(r = 0.998 8), 0.1–10 (r = 0.999 7), 0.1–10 (r = 0.999 8) and 0.1–10 μg/mL (r = 0.9998), respectively. The average
recovery rate of the method was over 82%, the intra- and inter-day precisions (RSD) were both smaller than 11%.
Alkylamide from Sichuan pepper was proved to be stable during sample storage, preparation and analysis periods.
Conclusion: This method is accurate, rapid and reproducible, and can be applied to determine the content of alkylamide
from Sichuan pepper in tissues rats administered with it.

Key words: alkylamide from Zanthoxylum bungeanum, high performance liquid chromatography (HPLC), rat, tissues