MSMS Tips No. 433
GC-TOFMS Application

Qualitative Analysis of Chemical Components in an Herbal Medicine
Product: Mass spectrometer (MS)Introduction
Herbal medicines contain multiple effective ingredients, making it difficult to study them systematically compared to Western drugs. Initial steps
in examining effective ingredients of an herbal medicine include qualitative analysis of chemical components contained in its raw material.
For this application, gas chromatography - mass spectrometer (GC-MS) is widely used. However, typical database search based on GC-MS often
fails to fully identify such chemical components.
JEOL’s GC-high resolution mass spectrometer (GC-HRMS), JMS-T2000GC, is capable of: 1) high precision mass analysis; 2) detection ofmolecular and fragment ions through electron ionization (EI) and soft ionization (SI); and 3) auto analysis of acquired data by msFineAnalysis AI
to determine chemical formulas and predict chemical structures. In this work, we identified the chemical components contained in an herbal
medicine using a JMS-T2000GC and msFineAnalysis AI.

Measurement
Dry wild chrysanthemum flowers (Flos Chrysanthemi Indici),a commercial herbal medicine, was used as a sample. Following thesteps shown in Figure 1, 1.1 g of the sample was immersed in 10 mL ofethanol (≥99.5%) and subjected to ultrasonic extraction at room
temperature for 30 minutes.
The supernatant of the extracted liquid was collected, added
anhydrous sodium sulfate, and let stand for 30 minutes.
The resulting solution was injected to the GC-HRTOFMS. EI and FI(Field Ionization) were used as ionization methods for GC-HRTOFMS
measurement, and msFineAnalysis AI was subsequently used foranalysis of the acquired data. Table 1 shows the measurement
conditions.

Results and Discussion
Figure 2 shows the total ion current chromatograms (TICCs) of EI andFI. A total of 141 components were detected, including eucalyptol,camphor, borneol, and bornyl acetate marked by ● in the figure.
These 4 components are reported to be active compounds(1).
For 81 components, their chemical formulas predicted from themolecular ions did not agree with the results of NIST database search.
However, msFineAnalysis AI contains an AI database of EI mass
spectra estimated from the chemical structures in addition to the NIST
database. The 81 components, when examined against the AI
database, turned out to be the compounds, which were in good
agreement with EI fragment ions patterns and had the chemical
formulas predicted by molecular ions. Figure 4 shows the results.
These peaks were identified primarily as terpenes, terpenoids, fatty
acids, alkanes, and sterols. The figure shows their groups anddetection ranges.

1. Added ethanol 10mL.
2. Sonication 30min.

SampleTable 1. Measurement conditions
GC-HRMS
Gas Chromatograph
Mode

Splitless

Inlet temperature

280 °C

ColumnDB-5MS, 30m x 0.25mm, 0.25μm(Agilent Technologies, Inc.)

Oven

50°C (1min) →10°C/min →320°C(10min)Carrier gas

He, 1.0mL/min

Injection volume1μL

TOFMS

JMS-T2000GC (JEOL Ltd.)

Ionization

EI+：70eV, 300μA
FI+：-10kV, 40mA

Monitor ion range

m/z 10-800

Analysis software

3. Took supernatant of ethanol solution.
4. Added anhydrous sodium sulfate.
5. GC-HRTOFMS measurement.

Sample preparation

8890 GC (Agilent Technologies, Inc.)msFineAnalysis AI (JEOL Ltd.)6. Analysis

JMS-T2000GCmsFineAnalysis

Measurement

Qualitative Analysis

Figure 1. Experimental process

Figure 2. EI and FI TICCs of Flos Chrysanthemi Indici ethanol extract.

Figures 3 and 4 respectively show the mass spectra and predicted chemical structure of the peak marked by ▼ in Figure 2, which wasdetected at a retention time of 21.96 minutes.
Its chemical formula was estimated to be C17H18O5 from the exact mass verified in the FI mass spectrum in Figure 3. NIST database searchfor the EI mass spectrum did not result in any compounds showing a similarity of 700 or more.
Meanwhile, Figure 4 shows the results of msFineAnalysis AI structural analysis of (2R)-4,7-dimethyl-1-propan-2-yl-1,2,3,5,6,8ahexahydronaphthalen-2-ol. The AI-predicted mass spectrum and measured mass spectrum of the compound have high levels of AI score andAI probability, providing effective structural information.

EI

M＋・FI

Figure 3. EI and FI mass spectra of the component at a retention time of 21.96 minutes
in Flos Chrysanthemi Indici ethanol extract.

Measured

PredictedFigure 4. Structural Analysis results of the component at a retention time of 21.96 minutes bymsFineAnlaysis AI.
SummaryThe results of this work demonstrates the effectiveness of msFineAnalysis AI. It is a powerful and efficient tool for qualitative analysis,saving time and labor in chemical formula determination and chemical structure prediction. Accurate qualitative analysis of herbal
components is useful for discovery and quality control in the field of herbal medicine. The GC-HRMS system with msFineAnalysis AI is
expected to be used for wide-ranging qualitative analysis of chemical components in natural resources including herbal medicines.

References[1] Jiesheng Ye,. Chemical Papers, 2009, 63, 5, 506-511. DOI: 10.2478:-009 to -0056:-0