HPLC-ESI-MS/MS法测定连翘心中的8种化学成分

Simultaneous quantification of eight constituents in the seed of Forsythia suspense(Thunb.) Vahl by HPLC-MS/MS method

目的建立HPLC-ESI-MS/MS法同时测定连翘心中连翘酯苷A、连翘苷、连翘脂素、芦丁、槲皮素、异槲皮苷、阿魏酸和橙皮苷的量。方法色谱柱为Diamonsil C18柱(150 mm×4.6 mm,5μm),流动相为甲醇-0.1%甲酸水,梯度洗脱,体积流量800μL/min,进样量10μL;质谱采用电喷雾离子源(ESI源),负离子模式监测,源喷射电压-4 500 V,离子源温度650℃,定量分析采用多反应监测模式(MRM)。结果连翘心中8种成分在线性范围内峰面积与质量浓度均呈良好的线性关系,加样回收率为98.1%~101.8%,精密度RSD值小于0.84%。连翘酯苷A、连翘苷、连翘脂素和芦丁的量较高,平均质量分数分别为1 609.78、80.08、86.64和373.86μg/g,槲皮素、异槲皮苷、阿魏酸和橙皮苷的量较低。结论该法灵敏度高、专属性好、结果可靠,适用于连翘心中多组分的测定。

Objective To develop an HPLC-MS/MS method for the simultaneous determination of forsythoside A, phillyrin, forsythiaside, rutin, quercetin, isoquereitrin, femlic acid, and hesperidin in the seed of Forsythia suspense （Thunb.） Vahl. Methods Chromatographic separation was performed on a Diamonsil Cl8 column （150 mm×4.6 ram, 5 μm） with linear gradient elution of methanol and 0.1% formic acid at a flow rate of 800 μL/min, and the injection volume was 10 μL. Multiple-reaction monitoring （MRM） was employed with switching electrospray ion source polarity in negative mode. The ion spray voltage was set at -4 500 V and the turbo spray temperature was maintained at 650℃. Results All calibration curves showed good linearity within the test ranges. The average recoveries of the compounds ranged from 98.1% to 101.8%. The precisions （RSD） for the investigated components were less than 0.84%. The contents of forsythoside A, phillyrin, forsythiaside and mtin are higher than quercetin, isoquerciain, ferulic acid and hesperidin. And the average contents of forsythoside A, phillyrin, forsythiaside and mtin are 1 609.78, 80.08, 86.64 and 373.86μg/g, respectively. Condusion An efficient, rapid, and sensitive method is first established for the qualitation and quantification of eight major components in the seed of Forsythia suspense （Thunb.） Vahl. The satisfactory results demonstrate that the method could be applied as a reliable quality control method for the seed of Forsythia suspense （Thunb.） Vahl.