HPLC-DAD同时测定红花中4种亚精胺成分

Simultaneous determination of four coumaroylspermidine constituents in Carthamus tinctorius by HPLC-DAD

采用高效液相色谱(HPLC)的方法,以Agilent Zorbax Eclipse XDB-C18(4.6 mm×250 mm,5μm)为色谱分离柱,流动相为47%甲醇/水等度洗脱,洗脱流速为1.0 m L·min-1,柱温30℃,检测波长为270,280,290,300 nm多波长检测的色谱条件同时测定红花中4种亚精胺成分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),和N1,N5,N10-(E)-tri-p-coumaroyl-spermidine(4)的含量,进样体积为10μL。各成分之间分离度良好,4种亚精胺成分的质量浓度分别在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),0.005 0~0.100 4(r=0.999 8)g·L^(-1)内与峰面积呈良好的线性关系,加样回收率为98.61%~100.9%,RSD为2.3%~3.0%。表明所建立的方法快速简便、具有较好的重复性和稳定性,可用于红花中4种亚精胺成分的含量测定。

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-C18column（ 4. 6 mm × 250 mm,5 μm） eluted with 47% methanol in an isocratic program. The flow rate was 1 m L·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%（ RSD2. 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.