2.3.3 Capsule and herbage essential oil contents and chemical compositions
The amount of essential oil in drug herbage and dry capsule were determined by hydro Clevenger distillation device. 100 grams of sample for drug herbage and 50 grams of sample for dry capsule were used. The samples were ground in the blender and 10 times pure water was added, and distilled for 3h. Essential oil values (%, v/w) were calculated by volume over dry mater. The obtained essential oils were taken into dark colored flasks and stocked at 4oC in a refrigerator until they were analyzed (Damyanova et al., 2016).
The chemical components of the essential oil samples from four collection times were defined by gas chromatography (GC) and gas chromatography-mass spectrometry (GC/MS) analyses. The analyses were performed using a Shimadzu GC-2010 Plus and Shimadzu GC/MS-QP2010 Ultra instruments. The 0. 1mI oil sample was dissolved in 10ml hexane and shaken vigorously. It was kept in the dark for 1-2 hours. The sample was taken into vials and given to the device. The information about the chromatographic method is given below:
Column: RXI-5MS (0.25um X 30m X 0.25mm)
Scan range: 35-600 m/z
Split ratio: 30
Oven temperature: 60oC for 1 min followed by a temperature rise at a 4 oC/min rate to 250oC (held for 4 min).
Flow rate: 1.50 ml/min
The essential oil components were identified by comparing their mass spectra, retention times, retention indices, and relative to C5-C40 n-alkanes, the FFNSC 1.2 and W9N11.1 mass spectral library, and the literature (Babushok et al., 2011).