754 F. ASKARI F. SEFIDKON FLAVOUR ANDAND FRAGRANCE JOURNAL Flavour Fragr. J. 2006; 21: 754–756 Published online 17 May 2006 in Wiley InterScience (www.interscience.wiley.com) DOI: 10.1002/ffj.1619 Essential oil composition of Pimpinella affinis Ledeb. from two localities in Iran F. Askari* and F. Sefidkon Research Institute of Forests and Rangelands, PO Box 13185-116, Tehran, Iran Received 2 August 2004; Revised 13 February 2005; Accepted 1 March 2005 ABSTRACT: Essential oils were isolated by hydrodistillation from the stems plus leaves, inflorescences and seeds of Pimpinella affinis Ledeb. individually. The plants were collected from Khojir and Chaloos (north-east and north of Tehran province, respectively). The yields of stem plus leaf, inflorescence and seed oils for the Khojir samples were 0.04%, 1.98% and 5.33% w/w and for the Chalous samples were 0.37%, 1.74% and 4.05% w/w, respectively. Limonene is a major constituent in the inflorescence and seed oils of the Khojir (47.9% and 90.5%, respectively) and Chalous samples (37.8% and 70.8%, respectively), whereas it was found in the stem plus leaf oil of the Khojir and the Chalous samples in low amounts (1.4% and 0.8%, respectively). γ -Terpinen-7-al is the major constituent in the stem plus leaf and inflorescence oils of the Khojir (69.9% and 37.6%) and Chalous samples (72.8% and 49.1%), but was not found at all in the seed oils. Caryophyllene oxide (9.1%) was found in the stem plus leaf oil of the Khojir sample, and methyl eugenol (9.7%) and (E)nerolidol acetate (9.1%) in the seed oil of the Chalous sample. Copyright © 2006 John Wiley & Sons, Ltd. KEY WORDS: Pimpinella affinis; Umbelliferae; essential oil composition; limonene; γ-terpinen-7-al Introduction Pimpinella affinis [syn. P. reuteriana Boiss., P. griffithiana Boiss., P. ambigua W. D. Koch ex Wolff, P. multiradiata (Boiss.) Korov., P. korovinii R. Kamelin] presents in different regions of Iran, Iraq, Soiree and Israel. It is a biennial aromatic plant, 20–110 cm in height, with white umble inflorescences and ellipsoid fruits. It grows wild in the center and north of Iran and is more abundant in Chalous and Khojir.1,2 Both of these places are mountainous regions with cold weather. The Chalous region has a temperature range of −30.5 °C to 28.2 °C and an average rainfall of 531 mm; the Khojir region has a temperature range of −17.6 °C to 37.2 °C and an average rainfall of 420 mm (measured over a 20 year period). We have reported the essential oil composition of P. aurea and P. anisum, previously. The yields of the stem plus leaf, inflorescence and seed oils of P. aurea were 0.4%, 1.5%, and 2.0% w/w, respectively. The major constituents of the stem plus leaf oil were found to be 1,8-cineol and limonene (21.4%), viridiflorol (12.8%), α-pinene (11.5%) and kessane (10.5%); the major constituents of the flower oil were viridiflorol (32.5%) and β -bisabolene (29.5%); while the main constituents of * Correspondence to: F. Askari, Research Institute of Forests and Rangelands, PO Box 13185-116, Tehran, Iran. E-mail: fatemeh_askari2002@yahoo.com Contract/grant sponsor: Research Institute of Forests and Rangelands, Iran. Copyright © 2006 John Wiley & Sons, Ltd. the seed oil were β-bisabolene (50.8%) and viridiflorol (37.0%).3 The yield of oil seeds of P. anisum by steam distillation was 3.3% w/w, and 11 compounds were identified. The main compound was trans-anethol (>90%).4 There are some reports on essential oil analyses of other Pimpinella species in the literature.5–11 The oil yield of P. anisum seed (by supercritical fluid extraction) and P. serbica were respectively 3.13–10.67% and 2–3.2%. The major compound of P. anisum was trans-anethol (90%) and that of P. serbica was β-caryophyllene (47.1%).5,6 The main components of the aerial parts oil of P. aromatica (6.1% yield) were methyl chavicol (91.1%) and trans-anethol (7.2%).7 Methyl chavicol was also the major compound (82.6%) in the fruit oil of P. aromatica.8 The main compounds of fruit and leaf oils of P. cumbrae were α-bisabolol (39% and 53%) and δ-3carene (16% and 11%), while the major compounds of the stem oil were α-bisabolol (39%) and isokessane (10%).9 The yield of stem plus leaf oil of P. squamosa was 0.17–0.29% and the fruit oil yield 4.6–7.0%. The main compound of the whole plant and fruit oils was trans-anethol (54.5% and 29.5%, respectively).10 The main compounds of P. achilleifolia aerial parts oil (pre-flowering and flowering) were p-cymene (52.2%), limonene (9.3%), α-phellandrene (8.8%) and car-2-ene (9.2%).11 To the best of our knowledge, there are no previous reports concerning the volatile constituents of P. affinis. The aim of this study was to determine of the quantity and quality of oils from different plant parts of P. affinis Flavour Fragr. J. 2006; 21: 754–756 ESSENTIAL OIL OF PIMPINELLA AFFINIS 755 from two localities. For this purpose, the oil compositions of three parts of P. affinis were studied. Experimental Plant materials were collected from Khojir and Chalous (north-east and north of Tehran province, respectively) at the flowering stage in June/July and at the seed stage in August/ September, 2003. The fresh plants were dried at room temperature. Herbarium specimens have been deposited in the Herbarium of the Research Institute of Forests and Rangelands (TARI). The dried parts of the plants (80–100 g samples of each) were crushed to small particles. Dried the stems plus leaves (S&L; collected during the flowering stage), inflorescences (IF) and seeds (S) were hydrodistilled for 3 h in a Clevenger apparatus to produce the oil. Three distillations were performed for each oil, then mixed for analysis. The sample oils were dried over anhydrous sodium sulphate and stored in sealed vials at 4 °C before analysis. GC Analysis The oils from the (S&L), (IF) and (S) samples of P. affinis were analysed using a Shimadzu GC-9A gas chromatograph equipped with a DB-5 fused silica column (30 m × 0.25 mm i.d., film thickness 0.25 µm; J&W Scientific); oven temperature, held at 40 °C for 5 min and then programmed to 260 °C at a rate of 4 °C/min; injector and detector (FID) temperatures, 270 °C; carrier gas, helium at a linear velocity of 32 cm/s. Percentages were calculated by area normalization method without the use of response factor correction. The retention indices were calculated for all compounds using a homologous series of n-alkanes. GC–MS Analysis GC–MS analyses were carried out on a Varian 3400 GC–MS system equipped with a DB-5 fused silica column (30 m × 0.25 mm i.d., film thickness 0.25 µm; J&W Scientific); oven temperature programme, 50–260 °C at a rate of 4 °C/min; transfer line temperature, 270 °C; carrier gas, helium at a linear velocity of 31.5 cm/s; split ratio, 1:60; ionization energy, 70 eV; scan time, 1 s; mass range, 40–300 amu. Identification of Compounds The constituents were identified by comparison of their mass spectra with those in computer libraries (LIBR-TR and Wiley- Copyright © 2006 John Wiley & Sons, Ltd. 5) or with authentic compounds. The identifications were confirmed by comparison of their retention indices, either with those of authentic compounds or with data in the literature.12 Results and Discussion The oil yields from samples of (S&L), (IF) and (S) of the Khojir samples were 0.04%, 1.98% and 5.33%, respectively, and those from similar samples of the Chalous samples were 0.37%, 1.74% and 4.05%, respectively. The oil yields of the generative parts (especially the seeds) were greater than the vegetative parts. The Khojir sample oils were light green and the Chalous sample oils were darker green. Eight constituents in the (S&L) oil, eight in the (IF) oil and nine in the (S) oil of the Khojir samples were identified. Ten constituents in the (S&L) oil, 13 in the (IF) oil and 10 in the (S) oil of the Chalous samples were identified (Table 1). In the Khojir samples, the major constituents of the (S&L) oil were γ -terpinen 7-al (69.9%), caryophyllene oxide (9.1%), aromadendrene (5.4%) and daucene (5.0%). The major constituents of the (IF) oil were limonene (47.9%) and γ -terpinen-7-al (37.6%). The major constituent of the (S) oil was limonene (90.5%). In the Chalous samples, the major constituents of the (S&L) oil were γ -terpinen-7-al (72.8%), cis-thujopsene (4.9%) and α-trans-bergamotene (4.7%). The major constituents of the (IF) oil were limonene (37.8%) and γ terpinen-7-al (49.1%). The major constituents of the (S) oil were limonene (70.8%), methyl eugenol (9.7%) and (E)-neridolol acetate (9.1%). Ten compounds (especially three major constituents) were common in the sample oils of two localities. Limonene was common to all the oils of both samples. γ Terpinen-7-al was not found in the seed oil. Limonene was found only in the seed oils of P. achilleifolia (9.29%) and P. cumbrae (8.0%).3,11 Comparison of these results with those reported previously3–11 showed that the oils of all parts of P. affinis have a specific composition. It seems that the high amount of limonene and γ -terpinen-7-al is a characteristic parameter for P. affinis oil. Of course, a literature review of Pimpinella oils showed sharp differences between the oil compositions of different species. Acknowledgements—The authors would like to acknowledge financial support provided by the Research Institute of Forests and Rangelands for this work. We thank Dr Mirza and MS Barazandeh for injection of the oils to GC–MS and GC, and Dr Mozafarian for identification of this species. Flavour Fragr. J. 2006; 21: 754–756 756 F. ASKARI AND F. SEFIDKON Table 1. Percentage of volatile compounds identified in the oil of Pimpinella affinis Ledeb. Compounds Sabinene Myrcene α-Terpinene Limonene Linalool cis-Limonene oxide trans-Limonene oxide Geijerene trans-Carveol cis-Carveol Carvone γ -Terpinen-7-al Pregeijerene α-Copaene Daucene Methyl eugenol (Z )-α-Bergamotene β -Caryophyllene cis-Thujopsene (E )-α-Bergamotene Aromadendrene β -Humulene Germacrene D Bicyclogermacrene Elemicine Spathulenol Caryophyllene oxide (Z )-Nerolidol acetate Humulene (14-hydroxy-α) (E )-Nerolidol acetate Zerumbene Total identified Khojir Chalous RI* Stem + leaf (%) Inflorescence (%) Seed (%) Stem + leaf (%) Inflorescence (%) Seed (%) 967 992 1014 1029 1098 1133 1138 1143 1216 1228 1240 1278 1286 1370 1379 1395 1413 1417 1428 1433 1433 1438 1472 1492 1552 1574 1577 1675 1709 1714 1731 — — — 1.4 — — — 3.5 — — — 69.9 — — 5.0 — — 1.8 — — 5.4 — 2.2 — — — 9.1 — — — — 98.3 2.5 — — 47.9 — — — 2.0 — — — 37.6 — — — 2.2 — — — — 1.2 — — — — — — — 4.2 — 2.4 100.0 2.6 0.7 — 90.5 — — 0.4 — — — — — 1.2 0.3 — 2.3 t 1.1 — — — — — — 0.8 — — — — — — 99.0 — — 0.5 0.8 — — — 3.4 — — — 72.8 — 1.1 — — — — 4.9 4.7 — 0.7 1.7 — — 1.1 — — — — — 94.0 — 0.2 — 37.8 0.3 2.1 1.0 3.3 0.9 0.5 1.0 49.1 — 0.4 — — — — 1.6 — — — — — 0.1 — — — — — — 98.4 2.1 t — 70.8 — — — — — — — — — — — 9.7 t — — t — — — t 2.1 — — 2.2 — 9.1 — 96.0 * Retention indices calculated on DB-5 column. t, trace, <0.05%. 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