Abstract
Limitations in the separation ability of conventional liquid chromatography system remains a challenge in developing a versatile method for simultaneously determining both hydrophilic and lipophilic constituents in herbal medicines (HMs). To measure compounds covering a broad polarity span in HMs, we developed a directly-coupled reversed-phase and hydrophilic interaction liquid chromatography-tandem mass spectrometry system. Samples were firstly separated according to lipophilicity by using a C18 column. Utilizing a T-piece as connector, the eluent was then pumped into an amide column to get further separation that mainly based on the hydrogen bonding effects. Dan-Qi pair, an extensively used herb-combined prescription in China, was selected to test the practicability and performance of the established system. A total of 27 components, containing 9 hydrophilic and 18 lipophilic constituents, were simultaneously determined using a schedule multiple reaction monitoring method in 15 min. Up to 69.9% content could be monitored in one injection in Dan-Qi pair extract, showing a significant advantage over previous methods. The proposed method was expected to benefit the controllability of herbal medicines.
Highlights
Herbal medicines (HMs) are usually derived from one or several crude materials and thereby contain numerous hydrophilic and lipophilic constituents[1,2,3,4]
Liquid chromatography coupled with tandem mass spectrometry (MS/MS) has been established as an important tool for the quantitative analysis of multiple non-volatile molecules in HMs13
Owing to the approachable construction of devices and good repeatability attributing to the online system, direct-coupled RP-hydrophilic interaction chromatography (HILIC) system is potential to be a powerful tool in quantitative analysis of samples with complex components, e.g., human urine[19], toxins in marine harmful algae[20], phospholipids[27], and Cistanche tubulosa[28]
Summary
Herbal medicines (HMs) are usually derived from one or several crude materials and thereby contain numerous hydrophilic and lipophilic constituents[1,2,3,4]. The directly-coupled RP-HILIC combination and valve switching-based assembly are both on-line systems. The former one is typically connected using a T-piece, which can bring in another mobile phase corresponding to the subsequent separation. This type of system is simple to set up and implement, but sacrifices some resolution and sensitivity compared with each single mode[25]. At least 8 preparations of this prescription are officially registered in Chinese Pharmacopoeia[31, 32], and one of them
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