10 mg of homogenous sample was scaled and diluted with 1 ml of pentane containing 0.04 % of decane as internal standard. The tube containing the sample solution was placed in ultrasonic bath for 5 min and then mixed. 200 µL of prepared solution was diluted with 800 µL of pure pentane mixed and individually analysed by GC-FID.
An Agilent HP 6890 gas chromatograph equipped with FID was used for the analysis of terpenoids. Separation was accomplished on a Rtx-5 w/Integra-Guard capillary column (30 m length, 0.25 mm i.d. and 0.25 µm df). Injections were carried out in split mode using a general purpose split/splitless liner packed with glass wool. The program started at 50 °C, increased to 280 °C (at 15 °C/min) and held for 15 min for a total of 31 min. 2 µL of each sample was injected with helium as the carrier gas (constant flow mode, 1 ml/min) using a split ratio 1:10. Temperatures were applied 280 °C for the injector, 260 °C for the transfer line. Data was analysed using Chemstation v.D.02.00.275 (Agilent Technologies).
List of the target Terpenes:
Decane (IS) 5,165
1. Decane concentration in pentane 0,04% = 1 ml pentane contain 0.292 mg decane.
2. 0.01g of sample dissolved with 1000 µL pentane containing 0.04 % decane.
3. Extract with concentration of 10 mg/ml dissolved 5 times. Final concentration of the extract in solution 2.5 mg/ml (0.0025 g/ml).
4. Final concentration of decane in solution 0.073 mg/ml.
5. 0.073 mg of decane / 0.0025 g of extract = 29.2 mg/g extract
Peak Area of decane - 29.2 mg/g extract
Peak area of target compound - x mg/g extract
x= 29.2 × Peak area of target compound / Peak area of decane = amount of target compound mg/ g extract.
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