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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 mobile phase was eluted with isocratic 47% methanol / water using a high performance liquid chromatographic (HPLC) method with an Agilent Zorbax Eclipse XDB-C18 (4.6 mm × 250 mm, 5 μm) column with an elution flow rate of 1.0 m L · min-1, the column temperature was 30 ℃, the detection wavelength was 270,280,290,300 nm, and the determination of four kinds of spermidine components in safflower rhizomes (N1, N5, N10- (Z) -tri-p-coumaroylspermidine N1-N5- (Z) -N10- (E) -tri-p-coumaroylspermidine (3) ), And the content of N1, N5, N10- (E) -tri-p-coumaroyl-spermidine (4) The separation between each component was good. The concentrations of the four spermidine components were 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). The linear range of the peak area was within the range of 0.005 0 ~ 0.100 4 (r = 0.999 8) g · L ~ (-1). The average recoveries were 98.61% ~ 100.9% with RSDs of 2.3% 3.0%. The results showed that the established method was rapid and simple, with good repeatability and stability, which could be used to determine the content of four kinds of spermidine in safflower.