Citation: LIU Yong-Li , WANG Chang-Shun , WANG Xiao-Lei , YANG Shu-Nan , LIU Xing-Guo , FENG Li . Simultaneous Quantification of Morroniside, Loganin, Peoniflorin, Paeonol in Mingmudihuang Pill by Online Two-dimensional Column Switching Ultra Performance Liquid Chromatography. Chinese Journal of Analytical Chemistry, 2016, 44(5): 828-832. doi: 10.11895/j.issn.0253-3820.150780 [复制]
Simultaneous Quantification of Morroniside, Loganin, Peoniflorin, Paeonol in Mingmudihuang Pill by Online Two-dimensional Column Switching Ultra Performance Liquid Chromatography
建立了二维柱切换-超高效液相色谱法同时测定明目地黄丸中莫诺苷、马钱苷、芍药苷、丹皮酚含量的方法。一维色谱柱为Thermo Accucore XL C18(250 mm×2.1 mm,4μm),二维色谱柱为DIONEX Acclaim phenyl-1(150 mm×4.6 mm,3μm),一维分析流动相为乙腈-水,梯度洗脱,二维分析流动相为乙腈-水,等度洗脱,14.5 min进行阀切换;检测波长:0-14.5 min为240 nm,14.5-30 min为275 nm,流速:0.5 mL/min,柱温:30℃。30 min即可完成明目地黄丸中莫诺苷、马钱苷、芍药苷、丹皮酚的含量测定,并有效地将莫诺苷同分异构体分离。莫诺苷、马钱苷、芍药苷、丹皮酚的线性范围分别为7.6~377 mg/L,9.2~459 mg/L,8.4~419 mg/L和8.2~409 mg/L,相关系数为0.9999,加样回收率为98.3%~100.2%。本方法快捷高效,可对控制明目地黄丸质量提供参考。
An online Two-dimensional column switching ultra performance liquid chromatography (2D-UPLC) method was established for simultaneous quantification of morroniside, loganin, peoniflorin, paeonol in Mingmudihuang Pill. The first-dimensional column was Thermo Accucore XL C18 (250 mm×2.1 mm, 4 μm) while the second-dimensional column was DIONEX Acclaim phenyl-1 (150 mm×4.6 mm, 3 μm). Acetonitrile and water were used as mobile phase with a gradient elution in the first-dimensional analysis and the second-dimensional analysis used acetonitrile-water as mobile phase with a isocratic elution. The valve switching time was 14.5 min, the detection wavelength was set at 240 nm in 0-14.5 min and 275 nm in 14.5-30 min. The flow rate was 0.5 mL/min, and the column temperature was 30℃. The determination of four compounds in Mingmudihuang Pill was completed in 30 min and it could effectually segregate the isomer of morroniside. The linear ranges of morroniside, loganin, peoniflorin and paeonol were 7.6-377 mg/L, 9.2-459 mg/L, 8.4-419 mg/L and 8.2-409 mg/L, respectively, and all the correlation coefficients (r) were>0.9999. The recoveries of the four compounds were 98.3%-100.2%. It was proved that this method could greatly improve the sufficiency of sample analysis and could control the quality of Mingmudihaung Pill efficiently.