Determination of Eucalyptol and Borneol in Mugwort Leaves By Wayeal Gas Chromatograph

As a traditional Chinese medicinal herb, the medicinal value of artemisiae argyi folium primarily depends on the content of its volatile components. Eucalyptol and borneol are two key active substances among them. Eucalyptol exhibits pharmacological effects such as anti-inflammatory, cough suppression, and asthma relief, while borneol possesses functions like clearing orifices and reviving consciousness, as well as reducing swelling and alleviating pain. The content of these components is susceptible to factors such as the geographical origin, harvesting season, and processing techniques of mugwort leaves. Therefore, accurate determination of their content holds significant value for ensuring the quality, efficacy, and safety of mugwort leaves as a medicinal material.

This study, with reference to the relevant methods outlined in the 2025 edition of the Chinese Pharmacopoeia, utilized the Wayeal gas chromatograph GC6100 equipped with a hydrogen flame ionization detector (FID) to determine the content of eucalyptol and borneol in mugwort leaves.

Keywords: Mugwort leaves; Eucalyptol; Borneol; Gas chromatography; FID detector.

1. Experiment Method

1.1 Instrument Configuration

Table 1 Configuration List of Gas Chromatograph

1.2 Experiment Materials and Auxiliary Equipment

Eucalyptol reference standard;

Borneol reference standard;

Ethyl acetate;

Volatile oil determination apparatus;

Carrier gas: High-purity nitrogen;

Hydrogen generator;

Air generator.

1.3 Test Conditions

Gas chromatography conditions

Column: DB-17 capillary column, 30m×0.25mm×0.25μm

Temperature programming: initial column temperature 45°C, increased to 75°C at a rate of 2°C/min, held for 5 minutes, then increased to 90°C at a rate of 1°C/min, held for 6 minutes, increased to 150°C at a rate of 5°C/min, then increased to 250°C at a rate of 10°C/min, and held for 5 minutes.

Column flow rate: 0.6mL/min

Inlet temperature: 240°C

Detector temperature: 250°C

Air flow rate: 300mL/min

Hydrogen flow rate:40mL/min

Make-up flow rate: 10mL/min

Split injection: Split ratio 5:1

Injection volume: 1μL

1.4 Solution Preparation

Preparation of Reference Standard Solution

Weigh 20mg of eucalyptol reference standard and 10mg of borneol reference standard into a 100mL volumetric flask, and add ethyl acetate to prepare a mixed solution containing 0.2mg of eucalyptol and 0.1mg of borneol per 1mL.

2. Result and Discussion

2.1 Qualitative Test of Standard Solution

Fig 1 Solution Chromatogram

Fig 2 Reference Standard Solution Chromatogram

Table 2 Chromatography Parameters of Reference Solutions

Note: As shown in the chromatogram above, the theoretical plate number of the borneol peak is greater than 50,000, which meets the requirements for experimental analysis.

2.2 Repeatability

Fig 3 Reproducibility Chromatogram of Reference Standard Solution

Table 3 Reproducibility Chromatographic Parameters of Eucalyptol

Table 4 Reproducibility Chromatographic Parameters of Borneol

Note: After five consecutive injections of the reference standard solution, the relative standard deviation (RSD) values for the retention times of both eucalyptol and borneol were less than 0.050%, and the RSD values for their peak areas were less than 0.8%, indicating good reproducibility.

2.3 Sample Test

Sample preparation: Take an appropriate amount of mugwort leaves and cut them into approximately 0.5cm fragments. Accurately weigh 2.5584g of mugwort leaves into a round-bottom flask, add 300ml of water, and connect to the volatile oil determination apparatus.

Add water from the top of the determination apparatus until the graduated section is completely filled and water overflows into the flask, then add 2.5ml of ethyl acetate and connect the reflux condenser. Heat the mixture to boiling and maintain heating for 5 hours. Allow it to cool, then separate the ethyl acetate layer and transfer it into a 10mL volumetric flask. Rinse the determination apparatus and condenser tube several times with ethyl acetate, combine the washings into the same volumetric flask, dilute to the mark with ethyl acetate, and mix well to obtain the final solution.

Fig 4 Sample Test Chromatogram

Table 5 Chromatographic Parameters of Eucalyptol in Mugwort Leaf Sample Solution

Table 6 Chromatographic Parameters of Borneol in Mugwort Leaf Sample Solution

Table 7 Volatile Oil Content in Mugwort Leaf Sample

Note: Calculated on a dried basis, the mugwort leaf sample contains 0.10% eucalyptol and 0.032% borneol, which complies with the requirements of the pharmacopoeia.

3. Conclusion

This test employed the Wayeal gas chromatograph GC6100 equipped with an FID detector to analyze eucalyptol and borneol in the mugwort leaf sample. The experimental results indicate that the theoretical plate number of the borneol peak in the reference standard solution is greater than 50,000, meeting the requirements for experimental analysis. After sample pretreatment and calculation on a dried basis, the mugwort leaf sample contains 0.10% eucalyptol and 0.032% borneol, which complies with the requirements of the pharmacopoeia. This demonstrates that the method, equipped with the Wayeal’s GC6100, fulfills the detection requirements for eucalyptol and borneol in mugwort leaf samples.