HPLC同时测定隔山消中5个成分的含量

目的: 建立同时测定隔山消中5个成分含量的高效液相色谱方法。方法: 采用Synergi 4μ Hydro-RP 80A色谱柱(4.6 mm×250 mm,4 μm),以甲醇-0.1%磷酸水溶液为流动相,梯度洗脱,流速1.0 mL·min^-1,检测波长260 nm(3-羟基-4-甲氧基苯甲酸、2,5-二羟基苯乙酮)、280 nm(4-羟基苯乙酮、白首乌二苯酮、2,4-二羟基苯乙酮),柱温35 ℃。结果: 3-羟基-4-甲氧基苯甲酸、2,5-二羟基苯乙酮、4-羟基苯乙酮、白首乌二苯酮和2,4-二羟基苯乙酮线性范围分别为0.010 72~0.343 0 μg(r=0.999 9)、0.005 470~0.175 0 μg(r=0.999 9)、0.011 04~0.353 3 μg(r=0.999 9)、0.042 80~1.370 μg(r=0.999 9)和0.021 76~0.696 3 μg(r=0.999 9);平均加样回收率(n=6)分别为96.9%、100.3%、97.1%、101.4%和97.9%。32批样品中上述5个成分的含量范围分别为12.90~272.4、6.00~171.3、4.200~661.8、12.47~1439、7.300~683.1 μg·g^-1。结论: 经方法学验证,该方法可用于隔山消药材的质量控制。

Objective: To establish an HPLC method for simultaneous determination of 3-hydroxy-4-methoxybenzoic acid, 4-hydroxyacetophenone, 2, 5-dihydroxyacetophenone, baishouwubenzophenone and 2, 4-dihydroxyacetophenone in Cynanchum auriculatum.Methods: The five constituents were measured on a Synergi 4μ Hydro-RP 80A(4.6 mm×250 mm, 4 μm)column with a gradient solvent system of methanol-0.1% phosphoric acid as the mobile phase at a flow rate of 1.0 mL·min^-1 and the column temperature was kept at 35 ℃.The detection wavelength was set at 260 nm for detecting 3-hydroxy-4-methoxybenzoic acid, 2, 5-dihydroxyacetophenone as well as 280 nm for detecting 4-hydroxyacetophenone, baishouwubenzophenone, and 2, 4-dihydroxyacetophenone.Results: The linear ranges of 3-hydroxy-4-methoxybenzoic acid, 2, 5-dihydroxyacetophenone, 4-hydroxyacetophenone, baishouwubenzophenonea and 2, 4-dihydroxyacetophenone were 0.010 72-0.343 0 μg(r=0.999 9), 0.005 470-0.175 0 μg(r=0.999 9), 0.011 04-0.353 3 μg(r=0.999 9), 0.042 80-1.370 μg(r=0.999 9), and 0.021 76-0.696 3 μg(r=0.999 9), respectively.The average recoveries(n=6)of above five components were 96.9%, 100.3%, 97.1%, 101.4% and 97.9%.The contents of above five components in 32 samples were in the range of 12.90-272.4, 6.00-171.3, 4.200-661.8, 12.47-1 439, and 7.300-683.1 μg·g^-1 respectively.Conclusion: It is proved that the method can be used to control the quality of C.auriculatum.