ABSTRACT
The HPLC-DAD method was established to simultaneously determine the contents of four coumaroylspermidine[ N1, N5, N10-(Z)-tri-p-coumaroylspermidine (1), N1, N5-(Z)-N10-(E)-tri-p-coumaroylspermidine (2), N1(E)-N5-(Z)-N10-(E)-tri-p-coumaroylspermidine (3), and N1, N5, N10-(E)-tri-p-coumaroyl-spermidine (4) ] in Carthamus tinctorius. The method was performed on an Eclipse XDB-C18 column (4.6 mm×250 mm, 5 μm) eluted with 47% methanol in an isocratic program. The flow rate was 1 mL•min⁻¹; the injection volume was 10 μL, and the column temperature was 30 ℃. The detective wavelength was 270, 280, 290, and 300 nm, respectively. Four coumaroylspermidine constituents showed a good linearity in the range of 0.002 1-0.041 6 (r=0.999 5), 0.002 6-0.051 2 (r=0.999 7), 0.002 7-0.054 0 (r=0.999 8) g•L⁻¹, and 0.005 0-0.100 4 (r=0.999 8) g•L⁻¹, respectively. The average recoveries of these four coumaroylspermidine constituents were in the range of 98.61%-100.9% (RSD 2.3%-3.0%). In conclusion, the method is simple, rapid, and sensitive, which could be used as a quantitative determination method for the four coumaroylspermidine components in C.tinctorius.