SHORT COMMUNICATION VOLATILE CONSTITUENTS OF HAPLOPHYLLUM BUHSEI BOISS. FLOWERING AERIAL PARTS

The essential oil composition of flowering aerial parts of Haplophyllum buhsei Boiss. (Rutaceae) from Iran was analyzed for the first time using gas chromatography (GC) and gas chromatography–mass spectrometry (GC–MS). Thirty six compounds comprising 92.2% of the total oil were characterized. The main components were -caryophyllene (12.9%), limonene (9.7%), -pinene (7.9%), linalool (7.4%), α-pinene (6.4%) and 1,8-cineole (5.5%). Volatile oil of Haplophyllum buhsei Boiss. flowering aerial parts was dominated by the high content of monoterpene hydrocarbons (39.2%) and oxygenated monoterpenes (23.2%).


INTRODUCTION
The genus Haplophyllum A. Juss.belongs to Rutaceae family and encompasses approximately 70 species which mainly distributed around the Mediterranean region of Europe and through western Asia up to Siberia [1,2].Eighteen species of Haplophyllum A. Juss.genus are found in Iran which among them nine species are endemic and Haplophyllum buhsei Boiss. is one of them [3].
Plants of Haplophyllum A. Juss.genus have long been used in folk medicine for toothache, stomach and skin diseases, and in the treatment of some types of cancer.The extracts of some Haplophyllum species exhibit cytotoxic activity [4].This genus has been shown to possess alkaloids, lignans, flavonoids, coumarins and essential oils with important biological properties [5].
A thorough literature review revealed that volatile constituents of many species of the genus Haplophyllum A. Juss.have been widely investigated in various regions from all over the world [6][7][8][9][10][11][12][13][14][15][16][17] but to the best of our knowledge, there was no report on Haplophyllum buhsei Boiss.and in this article essential oil composition of this endemic species is informed for the first time.GC and GC-MS analyses.Chemical composition of the volatile oil was investigated by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS).Haplophyllum buhsei Boiss.oil was analyzed by GC-MS using a Hewlett-Packard 6890 gas chromatograph with DB-5 capillary column (30 m х 0.25 mm; film thickness 0.25 μm).The carrier gas was helium with a flow rate of l mL/min.The column temperature was programmed from 60 o C to 220 o C at 6 o C/min.The gas chromatograph was coupled to a Hewlett-Packard 5973 mass selective detector.The MS was operated at 70 eV ionization energy.The retention indices were calculated by using retention times of n-alkanes that were injected after the essential oil at the same conditions.The components were identified by comparison of retention indices with those reported in the literatures and also by comparison of their mass spectra with the published mass spectra or Wiley library [18,19].Gas chromatography using flame ionization detection (GC-FID) analysis was carried out under the same experimental conditions with the same column as described for the GC-MS.The relative percentage of the identified compounds was computed from the GC peak area without applying correction factors.

RESULTS AND DISCUSSION
The dried flowering aerial parts of Haplophyllum buhsei Boiss.yielded 0.35% V/W of a pale yellow volatile oil.Thirty six compounds comprising 92.2% of the total oil were detected which are presented in Table 1.
Plant material.Flowering aerial parts of Haplophyllum buhsei Boiss.were collected in May 2013 from Dalahu mountain (Dalahu County, Kermanshah Province, Iran).Essential Oil Isolation.The air-dried crushed flowering aerial parts of Haplophyllum buhseiBoiss.were subjected to hydrodistillation using a Clevenger-type apparatus for 4 hour.The obtained essential oil was dried over anhydrous sodium sulphate and stored at 4-6 o C.

Table 1 .
The essential oil composition of Haplophyllum buhsei Boiss.aerial parts.