反相-亲水作用色谱-定制多反应监测法同步测定肉苁蓉多成分含量  

Simultaneous Determination of Multiple Components of Cistanches Herba Based on Reversed Phase-Hydrophilic Interaction Chromatography-Tailored Multiple Reaction Monitoring

作　　者：宋青青[1] 张珂 李婷 管朋维 龚兴成 许霞 李军[1,2] 屠鹏飞[1] 宋月林 SONG Qing-Qing;ZHANG Ke;LI Ting;GUAN Peng-Wei;GONG Xing-Cheng;XU Xia;LI Jun;TU Peng-Fei;SONG Yue-Lin(Modern Research Center for Traditional Chinese Medicine,School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 100029,China;Beijing Key Lab for Quality Evaluation of Chinese Meteria Medica,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院中药现代研究中心,北京100029 [2]北京中医药大学中药品质评价北京市重点实验室,北京100029

出　　处：《分析化学》2020年第11期1573-1582,I0035-I0045,共21页Chinese Journal of Analytical Chemistry

基　　金：国家重点研发计划项目(Nos.2018YFC1707300,2017YFC702400);国家科学自然基金项目(Nos.81773875,81530097);北京中医药大学青年科学家培育计划项目(No.BUCM-2019-QNKXJB006)资助。

摘　　要：建立了反相色谱-亲水作用色谱-定制多反应监测(RPLC-HILIC-tailored MRM)方法,对肉苁蓉中极性跨度大、含量差异大的27个化学成分的含量进行同步测定。利用RPLC与HILIC的在线直接偶联实现大、中、小极性化学成分的同步色谱保留和分离。Acquity UPLC HSS T3色谱柱(100 mm×2.1 mm,1.8μm)用于RPLC,Xbridge Amide色谱柱(150 mm×4.6 mm,3.5μm)用于HILIC,色谱柱间通过混合器引入稀释泵来解决色谱机制不一致的问题。泵A(0.1%甲酸-水)和泵B(乙腈)输送流动相至RPLC柱进行梯度洗脱,流速为0.15 mL/min;泵C(0.1%甲酸-水)和泵D(乙腈)以1.0 mL/min的流速输送流动相至混合器,改变RPLC柱洗脱液的组成,促进极性大的化学成分在HILIC柱中的保留。通过灵活调节在线碰撞能,提高微量成分的质谱响应,抑制常量成分的质谱响应,形成定制多反应监测模式。最终同步测定36批肉苁蓉中27个化学成分,包括氨基酸类、有机酸类、糖类、苯乙醇苷类、环烯醚萜类、木质素类等类型。方法检出限为0.0032~160μg/g,定量限为0.032~320μg/g,线性相关系数大于0.9929,加标回收率为74.7%~125.1%,相对标准偏差(RSD)为1.6%~13.7%。本方法线性范围宽、准确度高、适用性广,为肉苁蓉及其它中药化学成分的深入定量分析提供了有效手段。Reversed phase-hydrophilic interaction chromatography-tailored multiple reaction monitoring(RPLC-HILIC-tailored MRM)was developed for simultaneous determination of 27 components with wide polarity and wide range of content in Cistanches herba.RPLC was directly coupled with HILIC to achieve comprehensive retention of numerous compounds regardless of the polarity.Acquity UPLC HSS T3 column(100 mm×2.1 mm,1.8μm)and Xbridge Amide column(150 mm×4.6 mm,3.5μm)were employed for reversed phase separation and HILIC mode,respectively.To overcome the obstacle of incompatible mobile phases,dilution pumps were involved between two columns via a mixer.Pump A and pump B were responsible for delivering 0.1%formic acid and acetonitrile,respectively,into the RPLC column with a flow rate of 0.15 mL/min by gradient elution.Pump C and pump D,served as dilution pumps,introduced the same mobile phase as pump A and pump B into the mixer at a flow rate of 1.0 mL/min,which contributed to retention of polar compounds on HILIC column.For mass detection,tailored MRM with optimal collision energy was carried out by online energy-resolved MS to suppress mass response of abundant constituents,and to advance mass response of trace compounds.A total of 27 components in thirty-six batches of Cistanches Herba,such as amino acids,organic acids,phenylethanoid glycosides,lignans,and iridoids,were determined by RPLC-HILIC-tailored MRM.Satisfactory correlation coefficients were observed for all calibration equations with greater than 0.9929 over their corresponding concentration ranges.The limits of detection(LODs)were 0.0032-160μg/g and the limits of quantification(LOQs)were 0.032-320μg/g.The average recoveries at three spiked levels were in the range of 74.7%-125.1%with relative standard deviations(RSDs)of 1.6%-13.7%.The developed method showed wide linear range,good accuracy and wide applicability,and had the potential for large-scale quantitative analysis of traditional Chinese medicine.

关 键 词：反相色谱-亲水作用色谱 定制多反应监测模式 极性跨度 含量跨度 肉苁蓉 

分 类 号：O657.7[理学—分析化学] R284.1[理学—化学]