LC-UV和LC-MS在山楂叶多元酚类成分分析中的应用比较

目的:对LC-UV和LC-MS在山楂叶多元酚类成分分析中的应用进行方法学比较。方法:采用高效液相色谱并联双检测器(VWD和MSD)法。色谱柱为Lichrospher C18(250×4.6 mm I.D.,5μm);流动相A为乙腈,B为0.5%甲酸溶液,梯度洗脱,0-12 min A由11%-17%,12-30 min A由17%-18%,30-45 min A由18%-40%,45-60 minA由40%-100%;流速为1ml/min,三向分流阀分流进入MSD和VWD进行检测;柱温30℃;进样量10μl。结果:LC-MS灵敏度比HPLC法高10倍以上,在检测样品中微量成分时具有很大的优势;另外,LC-MS是根据tR和m/z两项指标来确定成分的类型,对于某些用LC-UV方法不能得到良好分离的组分或在LC-UV图谱上保留时间与对照品相近的其它成分,可以选用选择性离子检测(SIM)进行分析,能避免这些成分的干扰,因而具有高度的选择性和专属性。而LC-UV法操作简便,成本低,流动相选择面较广,因而分离度往往比LC-MS高,在检测样品中具有较好的紫外吸收、较好的分离度以及含量较高的成分时,LC-UV法以其高精密度和高稳定性完全可以达到LC-MS同样的检测结果。结论:LC-MS和LC-UV在山楂叶多元酚类成分的定量分析方面各有特色,可根据不同的检测指标选用不同的检测器。

The Comparison between LC-UV and LC-MS in Analysis of Polyphenolic Components from Leaves of Crataegus L.

OBJECTIVE: To compare the analytical method of LC-UV and LC-MS determination of major polyphenolic components in leaves of Crataegus L. METHODS: By high-performance liquid chromatography method with VWD and MSD, Lichropsher C18 column (250 x 4.6 mm I.D., 5 microm); mobile phase consisted of solvent A (acetonitrile) and solvent B (0.05% formic acid) ; elution profile was: 0-12 min 11% to 17% A in B (linear gradient), 12-30 min 17% to 18% A in B (linear gradient), 30-45 min 18% to 40% A in B (linear gradient), 45-60 min 40% to 100% A in B (linear gradient); flow rate was 1.00 ml/min, flow into MSD and VWD by diffluence, column temperature 30 degrees C and the injection volume 10 microl. RESULT: The sensitivity of LC-MS was 10 times more than that of LC-UV, so it is preponderance for microanalysis. Additionally, because LC-MS can identify the component by retention time (t(R)) and m/z, it has high selectivity and exclusion for the determined component. However, the method of LC-UV is simple; the cost is lower; the separate effect is better. So it is preponderance to determine the higher content component, which has better separate effect. CONCLUSION: LC-UV and LC-MS exhibited their own predominance for determination of major polyphenolic components in leaves of Crataegus L. So the detector should be selected according to the determined targets.