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PC-32

Advanced SWATH-MS for comprehensive profiling and retrospective

adulteration analysis in Danshen (Salvia miltiorrhiza)

1
1
2,3
Yi-Feng Zheng, Han-Ju Chien, Chen-Yuan Tseng, Wei-Chen Wang, Cheng-Yu Kuo,
#,1
1
Y a -Ting Pan, Chao-Yu Chen, Chien-Chen Lai* ,1,4,5,6
1
1

1 Institute of Molecular Biology, National Chung Hsing University, Taichung 402202, Taiwan.
2 Department of Biochemical Science and Technology, National Chiayi University, Chiayi
600355, Taiwan.
3 The Education and Research Center of Intelligent Technology for Food and Agriculture,
National Chiayi University, Chiayi 600355, Taiwan.
4 Graduate Institute of Chinese Medical Science, China Medical University, Taichung 404328,
Taiwan.
5 Advanced Plant Biotechnology Center, National Chung Hsing University, Taichung, 402202,

Taiwan.
6 Graduate Institute of Food Safety, National Chung Hsing University, Taichung, 402202,
Taiwan.
* E-mail: lailai@dragon.nchu.edu.tw

Abstract
The processing methods applied to Danshen (Salvia miltiorrhiza), a traditional Chinese
medicine (TCM), significantly influence the types and concentrations of its compounds. To
systematically profile bioactive compounds and retrospectively analyze adulterations in
Danshen, a robust and efficient analytical platform was established. This platform integrates
ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass
spectrometry (UPLC-QTOF-MS/MS), employing both sequential window acquisition of all
theoretical fragment ion spectra (SWATH) and multiple reaction monitoring (MRM) modes.
The developed method exhibited excellent linearity, precision, accuracy, and recovery for
compound profiling in Danshen. Moreover, the SWATH-MS dataset facilitated the retrospective
identification of three additional compounds—salvianolic acid A, lithospermic acid, and
przewaquinone A—and enabled the detection of adulterants, including hydroxyzine, an
antihistamine. This comprehensive SWATH-MS platform not only enables rapid preprocessing
and accurate quantification but also supports retrospective identification of bioactive
compounds and detection of adulterations. Its application holds significant promise for reducing
adulteration with Western medicines in Danshen and other TCMs.

Keywords: Salvia miltiorrhiza; UPLC-QTOF MS; Data-independent acquisition; SWATH-MS;
Retrospective analysis