UHPLC-MS/MS法测定不同周期蒙药文冠木枝和叶中8个成分的对比研究

Comparative study of eight components in branches and leaves of Xanthoceras sorbifolia Bunge at different growth stages by UHPLC-MS/MS method

目的:建立UHPLC-MS/MS测定方法测定蒙药文冠木枝和叶中儿茶素、表没食子儿茶素、芦丁、槲皮苷、表儿茶素、二氢杨梅素、杨梅苷和二氢槲皮素的不同周期含量对比研究。方法:采用WATER-SCORTECS反相C_(18)(100 mm×2.1 mm;1.6μm)色谱柱,以0.1%甲酸水溶液为流动相A,乙腈为流动相B,梯度洗脱(0~1 min,5%B;1~10 min,5%B→28%B;10~11 min,28%B→95%B;11~14 min,95%B;14~15 min,95%B→5%B),柱温40℃,流速0.3 mL·min^(-1),进样体积2μL,多反应监测(MRM)方法中选择负离子进行检测。结果:在测定的浓度范围内8个化学成分线性关系良好(r>0.9976),精密度、重复性和稳定性良好,平均加样回收率为97.4%~106.0%,RSD≤5.0%。6批叶样品中8个成分测定结果分别为0.090~0.904 mg·g^(-1)、0.093~2.258 mg·g^(-1)、0.001~0.005 mg·g^(-1)、0.530~6.176 mg·g^(-1)、0.158~1.561 mg·g^(-1)、0.002~0.056 mg·g^(-1)、4.008~10.218 mg·g^(-1)和1.049~16.990 mg·g^(-1),枝中8个成分测定结果分别为0.384~1.025 mg·g^(-1)、0.911~2.427 mg·g^(-1)、0.008~0.127 mg·g^(-1)、0.870~2.295 mg·g^(-1)、0.659~1.746 mg·g^(-1)、0.125~1.079 mg·g^(-1)、2.296~4.681 mg·g^(-1)和1.958~4.946 mg·g^(-1)。含量测定结果显示8个成分在文冠木不同部位中的含量存在较大差异,叶中杨梅苷、芦丁、槲皮苷随生长周期变化明显,可作为文冠木叶质量控制的主要指标。结论:该方法准确度和灵敏度高,且稳定性和重复性好,适用于蒙药文冠木药材中8个成分的同时检测,为枝和叶的质量控制提供实验基础。

Objective:To establish an UHPLC-MS/MS method for determining eight primary components(catechin,epigallocatechin,rutin,quercitrin,epicatechin,(+)-dihydromyricetin,myricitrin and dihydroqurcetin)in the young branches and leaves of Xanthoceras sorbifolia Bunge,a medicinal plant from Mongolia,and to compare their contents in samples at different growth stages.Methods:A Waters CORTECS C_(18)(100 mm×2.1 mm,1.6μm)chromatographic column was adopted using the mobile phase comprised of water containing 0.1%formic acid(A)and acetonitrile(B)with gradient elution(0-1 min,5%B;1-10 min,5%B→28%B;10-11 min,28%B→95%B;11-14 min,95%B;14-15 min,95%B→5%B)at a flow rate of 0.3 mL·min^(-1).The temperature of the column was set at 40℃.Injecting volume was 2μL.Detection was conducted using electrospray ionization(ESI)in negative ion mode with multiple reaction monitoring(MRM).Results:The linearity of the eight chemical components was found to be excellent in the tested concentration ranges,with correlation coefficients above 0.9976.Precision,repeatability and stability were satisfactory and the average recoveries were between 97.4%and 106.0%with RSDs≤5.0%.In six batches of leaves,contents of catechin,epigallocatechin,rutin,quercitrin,epicatechin,(+)-dihydromyricetin,myricitrin and dihydroqurcetin were in the ranges of 0.090-0.904 mg·g^(-1),0.093-2.258 mg·g^(-1),0.001-0.005 mg·g^(-1),0.530-6.176 mg·g^(-1),0.158-1.561 mg·g^(-1),0.002-0.056 mg·g^(-1),4.008-10.218 mg·g^(-1)and 1.049-16.990 mg·g^(-1),respectively.In six batches of young branches,the contents ranged from 0.384-1.025 mg·g^(-1),0.911-2.427 mg·g^(-1),0.008-0.127 mg·g^(-1),0.870-2.295 mg·g^(-1),0.659-1.746 mg·g^(-1),0.125-1.079 mg·g^(-1),2.296-4.681 mg·g^(-1)and 1.958-4.946 mg·g^(-1),respectively.The contents of eight components varied a lot in samples from different parts.The contents of myricitrin,rutin and quercitrin in the leaves exhibited noticeable changes with the growth cycle,suggesting their potential as quality control markers for leaves of Xanthoceras sorbifolia.Conclusion:The method is accurate,sensitive,stable,repeatable,and suitable for simultaneous determination of eight components in Xanthoceras sorbifolia Bunge,offering reference for quality control of its leaves and branches.