Detection of minor constituents in TongMai Granules using NP×RP off-line two-dimensional liquid chromatography coupled with orbitrap mass spectrometry

doi:10.5246/jcps.2020.11.072

Abstract

Abstract:

TongMai Granules is a Chinese medicine patent drug to treat cardiovascular diseases. It is prepared from the water extract of Dan-shen, Chuan-xiong and Ge-gen. The main compounds of TongMai Granules are isoflavone glycosides, isoflavones, and phenolic acids. However, it is difficult to detect low-polar compounds such as tanshinones and phthalides though they may also contribute to the therapeutic effect. In this work, an off-line two-dimensional chromatography method was developed to enrich the minor constituents in TongMai Granules. The extract was subjected to an SEP-PAK silica SPE column and eluted with petroleum ether, ethyl acetate–methanol (6:1, v/v), and methanol, successively. The three fractions were then analyzed using ultra-high performance liquid chromatography coupled with orbitrap mass spectrometry (UHPLC/orbitrap-MS). The isoflavones from Ge-gen, phenolic acids from Dan-shen and phthalides from Chuan-xiong were distributed in different fractions. A total of 131 compounds, including 59 from Ge-gen, 26 from Dan-shen, 43 from Chuan-xiong, and 3 common ones from Dan-shen and Chuan-xiong were tentatively identified. Among them, 26 compounds were identified by comparing with reference standards. This study set a good example for comprehensive analysis of the constituents in traditional Chinese medicine formula.

Key words: TongMai Granules, Off-line two-dimensional chromatography, NP×RP, UHPLC/orbitrap-MS

CLC Number:

R284

Supporting:

Supporting Information

Figure S1. LC/UV chromatograms for fractions of TongMai Granule extract eluted from a silica gel column.

Figure S2. LC/MS chromatograms for fractions of TongMai Granule extract eluted from an SEP-PAK silica SPE column.

Figure S3. LC/MS chromatograms for fractions of TongMai Granule extract eluted from an SEP-PAK silica SPE column using the optimized method.

Figure S4. LC/MS chromatograms for fractions of a blank control eluted from an SEP-PAK silica SPE column using the optimized method.

Figure S5. Repeatability for the SPE pretreatment method of TongMai Granules (n = 3).

Lulu Xu, Zhanpeng Shang, Yue Chai, Yang Zhang, Rong Yu, Min Ye, Xue Qiao. Detection of minor constituents in TongMai Granules using NP×RP off-line two-dimensional liquid chromatography coupled with orbitrap mass spectrometry[J]. Journal of Chinese Pharmaceutical Sciences, 2020, 29(11): 793-803.

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