fruticosa (Craib) J. Sinclair*

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1 «± µ π ªï Ë Ûapple Ë Ú..-.. ÚıÙ The Influence of Stages of Floral Development on Essential Oil Composition in Cananga odorata (Lam.) Hook.f. & Thomson var. fruticosa (Craib) J. Sinclair* W. Wongkaew 1 Associate Fellow of the Academy of Science, The Royal Institute, Thailand K. Chiangthong 2 Abstract The stages of floral development showed little effect on essential oil component in Cananga odorata (Lam.) Hook. f. & Thomson var. fruticosa (Craib) J. Sinclair. The flowers, however, emit different volatile compounds at different stages of development. The odour is more floral at the mature stage than the early one. This floral type is the one of commercial value. Further study is needed to rectify the contradicting result. Key words : essential oil, Cananga odorata var. fruticosa, stages floral development Introduction Thailand is enriched with plant diversity amounting to 11,000 species of vascular plants many of which produce essential oils. Essential oils are natural compounds found in parts of plants and animals. The world population is now moving into an era with increasing demand for naturally grown products with minimal use of synthetic chemicals both in agriculture and for other purposes. Therefore, essential oils are of potentially economic importance, especially in the spa business for health and beauty. However, some essential oils available in the spa market are still very expensive. Since most ylangylang oil is produced in Mardagascar, this study is part of a project to search for low-cost, essential oils-bearing plant such as Kradang-nga Songkhla which can be a potentially alternative crop for Thai farmers. This species is native to Thailand but unknown in the perfume industry, unlike the ylang-ylang or Kradang-nga Thai (Cananga odorata var. odorata) which yields pleasant odour suitable for commercial use. If Kradang-nga Songkhla (Cananga odorata var. fruticosa) can be an alternative crop for Kradang-nga Thai, it will be very convenient to cultivate and collect essential oil since Kradang-nga Songkhla is a shrub with easy-to-root * Presented at the meeting of the Academy of Sience, The Royal Institute on October 16, Department of Botany, Faculty of Science, Kasetsart University, Bang Khen Campus 2 Department of Biology, Rajabhat University, Kanchanaburi W. Wongkaew, K. Chiangthong 403

2 The Journal of the Royal Institute of Thailand Vol. 30 No. 2 Apr.-Jun cuttings which flowers readily as compared to the woody Kradang-nga Thai. Materials and Methods Plant Material Sixty out of 100 potted plants grown in 10 inches pot for 2 months were selected for experimental material. Plant cuttings were obtainable at Chatuchak market. Experimental Design Completely randomized design was used by lottery technique. Determination of flower number and fresh weight were done by tagging every flower to be counted and recorded on the first day of the month. At the end of the season, total number of flowers and fresh weight were calculated. Seasons were determined as summer (March-June), rainy season (July-October) and winter (November-February) Essential oil study Flowers were collected in the summer, rainy and winter for essential oil extraction which will be analysed by GC and GC-MS. Extraction was done by steam distillation, solvent extraction with dichloromethane, reagent grade. GC used in this experiment is CP (Chrompack) GC column was HP-5 fused silica capillary column (30 m Ó 0.25 mm. Ó 0.25 micromill.) produced by J and W Scientific Co, ltd. FID detector was used. Temperature program was set at 45 degree celsius increasing at the rate of 3 degree C/min from degree celsius and maintain temperature at 220 for 10 minutes. Injector temperature was set at 230 degree celsius while the detector was set at 240. Helium was the carrier gasat the rate of 2 ml/min. working on split mode at the ratio of 1:100. Injection volume was 1 ul. GC-MS used is the product of Varian model STAR 3400 Cx. with column HP-5 with the same temperature program as the one used in GC. Injector temperature was 230 and 270 for interface line. Carrier gas is helium at the rate of 2 ml/min., working as split mode in the ratio of 1:100. Injection volume was 0.3 ul. MS is SATURN 4 D made by varian directly coupled with GC. lon source was set at 230 and average electron energy was 70 ev. Identification of each compound of the essential oil was done by comparing the retention indices and mass spectra compared with NIST library computerized in the GC-MS set up. Determination of the quantity of the component was done by calculating percentage of peak area in GC chromatograms, computerized by normalization method from the equation. % A = area under peak Ó 100 total peak area Experimental Site Analysis was done in KU laboratory both at Kamphaeng Saen and Bang Khen Campus. Field work was done in Kanchanaburi. Result and Discussion The floral development in Cananga odorata or Kradang-nga Thai was first observed from floral bud to green stage in days and then from green to greennish yellow and finally yellow in about days, when the petal base developed a reddish spots. This is the stage ready for flower harvest (Weiss 1997), but the situation is different in Cananga odorata var. fruticosa which last days for full maturity ready to harvest. This differences may be due to genetics and/ or environmental factors. (Figure 13) Essential oil composition of Kradang-nga Songkhla seemed to exhibit little degree of differences, during stages of floral development. At the floral green, yellowish green, greenish yellow and yellow stages, the most abundant compound of the essential oils were mainly the same. The most abundant was α-farnesene. Other compounds with high amount are N-amylpyrrole, β- caryophyllene and germacrene D. Ding, et al (1988) also found similar essential oil composition. The highest one was β-caryophyllene followed by r-muurole, α-farnesene and α- humulene respectively. Hudaib, et al (2002) studied essential oil harvested from 2 year and 5 year old thyme in the same season, both yielded thymol at the highest level in June and July. Overton and Manura (2002) found that essential oil of chrysanthemum increased as flowers became more mature, perhaps due to increasing growth rate and as a consequence, increasing metabolism. 404 The Influence of Stages of Floral Development on Essential Oil Composition in Cananga odorata (Lam.) Hook.f. & Thomson var. fruticosa (Craib) J. Sinclair

3 «± µ π ªï Ë Ûapple Ë Ú..-.. ÚıÙ It is suggested here that Kradang-nga Songkhla yields essential oil in flower since the green stage and gradually emits the volatile oils towards the mature stage because of more accumulation of noticeable component. However, from the human perception the odour become more floral as the flowers mature. Further study is needed to rectify this difference. Reference Ding, J., Y. Yi, Y. Wu, Z. Lin and S. Dao Studies on the constituents of the essential oils from Cananga odorata in the different varieties and the flowered periods. Acta Botanica Yunnanica. 10 : Hudaib, M., Speroni, A.M.D. Pietra and V. Cavrini GC/MS evaluation of thyme (Thymus vulgaris L.) oil composition and variations during the vegetative cycle. J. Pharm. Biomed. Anal. 29 : Overton, S.V. and J.J. Manura Seasonal variation in flower volatiles. Available Source : http : //www. Sisweb.com/reference/applnote/ app-5.5htm, April 3, Weiss, E.A Essential Oil Crops. Cab international, New York. Figure 1 collected in summer at green stage. W. Wongkaew, K. Chiangthong 405

4 The Journal of the Royal Institute of Thailand Vol. 30 No. 2 Apr.-Jun Figure 2 collected in summer at yellowish green stage. Figure 3 collected in summer at the greenish yellow stage. 406 The Influence of Stages of Floral Development on Essential Oil Composition in Cananga odorata (Lam.) Hook.f. & Thomson var. fruticosa (Craib) J. Sinclair

5 «± µ π ªï Ë Ûapple Ë Ú..-.. ÚıÙ Figure 4 collected in summer at yellow stage. Figure 5 collected in the rainy season at green stage. W. Wongkaew, K. Chiangthong 407

6 The Journal of the Royal Institute of Thailand Vol. 30 No. 2 Apr.-Jun Figure 6 collected in the rainy season at yellowish green stage. Figure 7 collected in the rainy season at greenish yellow stage. 408 The Influence of Stages of Floral Development on Essential Oil Composition in Cananga odorata (Lam.) Hook.f. & Thomson var. fruticosa (Craib) J. Sinclair

7 «± µ π ªï Ë Ûapple Ë Ú..-.. ÚıÙ Figure 8 collected in the rainy season at yellow stage. Figure 9 collected in the winter at green stage. W. Wongkaew, K. Chiangthong 409

8 The Journal of the Royal Institute of Thailand Vol. 30 No. 2 Apr.-Jun Figure 10 collected in the winter at yellowish green stage. Figure 11 collected in the winter at greenish yellow stage. 410 The Influence of Stages of Floral Development on Essential Oil Composition in Cananga odorata (Lam.) Hook.f. & Thomson var. fruticosa (Craib) J. Sinclair

9 «± µ π ªï Ë Û Ë Ú..-.. ÚıÙ Figure 12 collected in the winter at yellow stage. W. Wongkaew, K. Chiangthong 411

10 The Journal of the Royal Institute of Thailand Vol. 30 No. 2 Apr.-Jun Figure 13 Photographs showing different stages of floral development of Kradang-nga Songkhla. 412 The Influence of Stages of Floral Development on Essential Oil Composition in Cananga odorata (Lam.) Hook.f. & Thomson var. fruticosa (Craib) J. Sinclair

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