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

引用本文: 宋青青, 张珂, 李婷, 管朋维, 龚兴成, 许霞, 李军, 屠鹏飞, 宋月林. 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[J]. 分析化学, 2020, 48(11): 1573-1582. doi: 10.19756/j.issn.0253-3820.201104 shu

Citation: SONG Qing-Qing, ZHANG Ke, LI Ting, GUAN Peng-Wei, GONG Xing-Cheng, XU Xia, LI Jun, TU Peng-Fei, SONG Yue-Lin. Simultaneous Determination of Multiple Components of Cistanches Herba Based on Reversed Phase-Hydrophilic Interaction Chromatography-Tailored Multiple Reaction Monitoring[J]. Chinese Journal of Analytical Chemistry, 2020, 48(11): 1573-1582. doi: 10.19756/j.issn.0253-3820.201104 shu

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

宋青青 ^1, ,
张珂 ^1, ,
李婷 ^1, ,
管朋维 ^1, ,
龚兴成 ^1, ,
许霞 ^1, ,
李军 ^1,2, ,
屠鹏飞 ^{1,
,} ,
宋月林 ^{1,2,
,}

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

通讯作者: 屠鹏飞,pengfeitu@163.com; 宋月林,syltwc2005@163.com

基金项目:
本文系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）和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，1.8 μm）用于RPLC，Xbridge Amide色谱柱（150 mm×4.6 mm，3.5 μm）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0.0032~160 μg/g，定量限为0.032~320 μg/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。

关键词:

English

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

1.
Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100029, China;
2.
Beijing Key Lab for Quality Evaluation of Chinese Meteria Medica, Beijing University of Chinese Medicine, Beijing 100029, China

Corresponding author: TU Peng-Fei, ; SONG Yue-Lin,

Received Date: 28 February 2020
Revised Date: 08 September 2020
Fund Project:
This work was supported by the National Key Research and Development Project (Nos. 2018YFC1707300, 2017YFC702400), the National Natural Science Foundation of China (Nos. 81773875, 81530097), the Young Elite Scientists Sponsorship Program by CAST (No. 2017QNRC001) and the Young Scientist Program by Beijing University of Chinese Medicine (No. BUCM-2019-QNKXJB006).

Abstract: 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, 1.8 μm) and Xbridge Amide column (150 mm×4.6 mm, 3.5 μm) 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 0.0032-160 μg/g and the limits of quantification (LOQs) were 0.032-320 μg/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.

Key words:

Reversed phase-hydrophilic interaction chromatography
/ Tailored multiple reaction monitoring
/ Polarity span
/ Content span
/ Cistanches herba

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沈阳化工大学材料科学与工程学院沈阳 110142

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

通讯作者: 屠鹏飞,pengfeitu@163.com; 宋月林,syltwc2005@163.com

English

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

Corresponding author: TU Peng-Fei, ; SONG Yue-Lin,

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

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

通讯作者: 屠鹏飞,pengfeitu@163.com; 宋月林,syltwc2005@163.com

English

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

Corresponding author: TU Peng-Fei, ; SONG Yue-Lin,

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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