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55 sources of heat and use of energy, the
drying method can be performed either by natural or artificial method. It reduces the enzymatic activity by inhibiting chemical reactions such as hydrolysis, oxidation, and fermentation (Costa et al., 2005). Many drying methods such as convection oven drying, freeze-drying, microwave drying etc. are also used to preserve medicinal herbs (Harbourne et al., 2009).
Ocimum americanum L. commonly called as Ocimum canum Sims. belonging to the family Lamiaceae [Wealth of India] is a wild herb with a distinct mint flavor, hairy leaves and scented flowers that is native to tropical Africa [Steel J. 2006 ] and is useful for its antimicrobial, antioxidant, antihelmintic and anti diabetic activities (Khare, C.P., 2007). Fresh lamiaceae herbs as spearmint usually contain 75–80%
water, and the water levels needs to be lowered to less than 15% for their successful preservation (Dìaz-Maroto et al., 2002). Drying may improve the herb quality such as appearance and aroma due to the loss of volatile compounds or the formation of new volatile compounds through desertification or oxidation reactions (Hossain et al., 2010).
(Nykanen and Nykanen, 1987) have reported that the total yield of essential oils decreased between 36–45% in sweet basil during drying at ambient temperature. Di Cesare et al., (2003) observed microwave drying to retain high percentages of characteristic volatile compounds (eucalyptol, linalool, eugenol, and
methyleugenol) in basil (Ocimum basilicum L.) compared to samples dried by air-drying and freeze-drying with blanching, except freeze-dried unblanched leaves.
Baritaux et al., (1992) found that the contents of methyl chavicol and eugenol decreased during drying, however, the levels of trans-bergamotene, linalool and 1, 8 - cineole significantly increased in basil (Ocimum basilicum L.). Some more reports are available on the effect of drying processes on Calendula essential oil from Nigeria (Okoh et al., 2008), Mentha from Norway, Nigeria and Egypt (Rohloff et al., 2005). In Mentha longifolia, the yield of dried plant material was more than that of fresh plant material (Asekun et al., 2006).
Shade dried and sun dried leaves of Plectranthus glandulosus showed difference in their chemical composition (Katamssadan et al., 2014). These investigations revealed that the chemical composition of oil varied with temperature and humidity conditions. Selection and optimization of drying methods could help in minimizing the loss of volatiles and also maintain the quality of the medicinal species. After surveying literature, it was observed that different drying methods like sun drying, shade drying, oven drying, fire heat drying etc. have been used to dry different aromatic and medicinal plants, but to the best of our knowledge, no work has been reported, on the impact of drying on the chemical composition of O.
americanum from India. Therefore, the
56 purpose of this study is to investigate the
effect of different drying on yield and chemical composition of O. americanum and find out the ways to obtain optimum active constituent condition.
Materials and Method Collection of plant material
Fresh O. americanum plants at flowering stage were harvested from the field of Ranikhet, Uttarakhand in September 2014.
A fraction of plant material was sun dried.
Botanical identification of plant was done at C.C.R.A.S. Thapla. Voucher no.
(24660).
Isolation of essential oil
Fresh and sun dried plant material was sliced into small parts and 500 gm of each sample was extracted by using hydro distillation technique in a glass Clevenger apparatus for about 4 hours. The extracted crude oil was stored in glass vials and excess moisture was dried over anhydrous sodium sulphate. The sealed glass vial was stored in BOD incubator.
Analysis of the essential oil
A Shimadzu 2010 auto system GC fitted with Rtx-5 MS and FID detector was used to analyze the oil. Carrier gas was the N2/air with linear velocity of 30.0 cm/sec.
Column temperature was set at 50°C for 2 min, then programmed until 210°C at a rate of 3°C/min with hold time of 2 min, and finally increased to 280°C at the 10°C/min rate with 11 min hold time, using N2 at 3 mL /min column head pressure as carrier gas, Injector and detector (FID, Flame ionization detector) temperatures were
260°C and 270°C respectively. Injection mode, split; split ratio 110.0, volume injected, 1 μl of the oil. The GC-MS used was Auto system 2010 GC (MS Serial Number: 1009701) capillary column (30 m
× 0.25 mm, 0.25 μm film thickness).
Column temperature was set at 50°C for 2 min, then programmed until 210°C at a rate of 3°C/min with hold time of 1 min, and finally increased to 280°C at the 10°C/min rate with 7 min hold time, using N2/Air as a carrier gas. Injector temperature was 260˚c and 1.0 uL in n- hexane. Split ratio 110.0 Relative percentage amounts of the essential oil components were evaluated from the total peak area (TIC) by apparatus software.
Identification of the components
Identification of components in the oils was done by comparing their mass spectra and retention time with literature data and by computer matching with NIST and WILEY library as well as by comparison of the fragmentation pattern of the mass spectral data with those reported in the literature (Adams, 2007).
Results and Discussion
The GC and GC/MS results revealed that there was a difference in the chemical composition of oils after drying. Oil yield of fresh and sun dried material was also significantly different .with values ranging from 0.10% for the fresh herb to 0.66%
(v/w) for the sun dried material. During drying process the amount of essential oil can decrease from 2.55% to 1.94% (Halva, 1987). Whereas, in our study, the yield of
57 essential oil from sun dried plant material
was greater than that from the fresh plant.
Total number of components in the oil of fresh and sun dried O. americanum were 48 and 116 respectively. The chemical composition of fresh aerial parts and sun dried of O. americanum is listed in Table 1.
The major compounds in fresh sample of O. americanum were bisabolene <β>
(29.06%), bisabolene <(E)-γ-> (17.49%), cineole <1,8-> (9.14%), methyl chavicol (7.56%), ocimene <(Z)-β-> (7.18%), germacrene D (5.80%) farnesene <(E)-β>
(4.09%) and caryophyllene <(E)->(2.61%).
The chemical characterization of O.
americanum showed the presence of camphor (60%) (Xaasan et al., 1981), trans-β-ocimene (29%) and 1, 8-cineole (41.3%) (Tchoumbougnang et al., 2006), citral (62.43%) (Saeio et al. 2011) and (E)- methyl cinnamate (95.8%) (Vieira and Simon, 2000).
After sun drying and the major compounds of the same were cineole <1,8-> (17.86%), camphor (38.34%), bisabolene <β>
(7.77%) and copaene <α-> (3.99%).
Our results showed that sun-drying caused significant loss of some major components like bisabolene <(E)>-γ-> (17.49% to 0%), methyl chavicol (7.56% to 0.01%), ocimene <(Z)-β-> (7.18% to 0%), eugenol (2.68% to 0%) and sesquisabinene (2.12%
to 0%). . Drying also brought about significant appearance of some components in oil after sun drying which were totally absent in the fresh oil like camphene (0%
to 1.93%) and 1-octen-3-ol (0% to 1.10%) in sun dried sample.
In Mentha longifolia, the yield of dried plant material was observed to be more as compared to fresh plant material. In a previous investigation, the oil of fresh plant material contained 92.6% of monoterpeinoids and major compounds were pulegene (35.0%), menthone (31.1%) and 1,8- cineole (13.0%). On the other hand, in sun dried plant material, percentage of monoterpenoid was 91.8%
and major compounds were menthone (38.3%), pulegone (20.2%) and 1,8-cineole (16.6%) (Asekun et al., 2006). Oven dried (45˚C) Rosemary resulted in loss of 7.25%
volatile components, while microwave drying causes 61.5% loss of volatile component in the same plant (Jaganmohan et al., 1998).
The present study indicated that there is loss of some components during drying whereas appearance of some components also occurs in drying. In O. americanum collected from Ranikhet, concentration of camphor increased significantly (0.08% to 38.34%). Fresh plant contains high concentration of methyl chavicol and linalool. Methyl chavicol is suspected to be carcinogenic and linalool is used as a scent in perfumed hygiene products and cleaning agents and also in mosquito repellent products, thus it is good to use the plant to obtain the above benefits. On the other hand, camphor has stimulant, antiseptic, decongestant, anesthetic and insecticidal properties and hence it is an important
58 constituent of many medicines and is used
for the treatment of cough, pain and skin irritation. The oil yield increased by drying and the quality of the volatile constituents
was also maintained thereby also increasing the commercial potential of the herb.
S. No. Name of compound RT RI
calculated RI Adams O. americanum
Fresh Sun-dried
1. Isovalerate <ethyl-> 4.39 822 849 0.57 2.17
2. Hex-(3Z)-enol 5.71 864 850 - 0.01
3. Thujene<α-> 7.70 936 924 - 0.05
4. Pinene<α-> 7.95 942 932 - 0.02
5. Camphene 8.54 958 946 - 1.93
6. Sabinene 7.96 984 969 0.17 0.01
7. Pinene<β-> 8.10 986 974 0.82 0.21
8. 1-octen-3-ol 9.77 992 974 - 1.10
9. 3-octanone 10.06 999 979 - 0.04
10. Myrcene 8.64 1002 988 - 0.56
11. Octanal<n-> 9.30 979 998 - -
12. Phellandrene-α 10.74 1015 1002 - 0.10
13. Hexenyl acetate<(3Z-> 10.94 1020 1004 - 0.05
14. Terpinene<α-> 9.67 989 1014 0.05 -
15. Cymene<p-> 10.05 1035 1020 0.04 -
16. Limonene 11.75 1039 1024 - -
17. Cineole<1,8-> 10.38 1041 1026 9.14 17.86
18. Ocimene<(z)-β-> 10.53 1011 1032 7.18 1.54
19. Ocimene<(E)-β-> 12.76 1059 1044 - 0.07
20. γ-Terpinene 13.14 1071 1054 - 1.28
21. Sibinene hydrate<cis-> 13.56 1080 1065 - 0.25
22. Mentha-2,4(8)-dinene<p-> 11.42 1037 1085 0.08 -
23. Linalool 13.85 1087 1095 0.28 0.35
24. Menth-2-en-1,ol<cis-para-> 14.93 1111 1118 - 0.07
25. Sabinol<trans-> 12.10 1046 1137 0.10 -
26. Camphor 15.29 1119 1141 0.08 38.34
27. Isoborneol 18.21 1183 1155 - 0.02
28. Terpineol(δ) 16.58 1119 1162 0.22 -
29. Borenol 18.10 1181 1165 - -
30. Menthol 18.49 1189 1167 - 0.03
31. Terpinen-4-ol 16.92 1192 1174 - 0.06
32. Terpinol<α-> 19.49 1208 1186 0.04 0.26
33. Methyl chavicol 17.88 1184 1195 - 0.01
34. Octanol acetate 20.20 1227 1211 7.56 -
35. Frenchyl acetate<endo-> 20.54 1235 1220 - -
36. Neral 21.06 1246 1235 - -
37. Geranial 22.84 1286 1264 - -
38. Anethole<(E)-> 23.50 1301 1284 - -
39. Copaene<α-> 27.44 1407 1345 - 0.02
40. Eugenol 24.81 1331 1356 2.68 -
41. Copaene<α-> 25.27 1343 1374 0.17 3.99
42. Bourbonene<β-> 25.60 1400 1387 0.65 0.08
43. Elemene<β-> 25.93 1358 1389 0.28 -
44. Methyl eugenol 28.78 1425 1403 - 0.06
45. Gurjunene<α> 28.82 1426 1409 - -
59
46. Caryophyllene<(E)-> 27.13 1385 1417 2.61 0.04
47. Cymene<2,E-dimethyl-p-> 29.49 1442 1424 - -
48. Copaene<β-> 29.67 1447 1430 - 0.02
49. Gurjunene<β-> 29.75 1449 1431 - -
50. Bergamotene<α-trans> 27.78 1453 1432 0.17 1.42
51. Sesquisabinene 30.29 1408 1440 2.12 -
52. Humulene<α-> 30.65 1471 1452 - 0.75
53. Farnesene<(E)-β-> 28.74 1424 1454 4.09 0.34
54. Caryophyllene<9-epi-(E)-> 30.94 1438 1464 - -
55. Germacrene-D 29.76 1484 1484 5.80 0.90
56. Bisabolene<β-> 31.24 1489 1505 29.06 7.78
57. Amorphene<δ-> 31.40 1472 1511 0.26 -
58. Cubebol 32.19 1534 1514 - -
59. Sesquiphellandrene<β-> 31.53 1492 1521 0.05 -
60. Cadiene<δ-> 33.51 1542 1522 - 0.03
61. Bisabolene<(E)-γ-> 32.47 1516 1529 17.49 -
62. Nerolidol 35.07 1583 1531 - -
63. Spathulenol 35.68 1598 1577 - -
64. Caryophyllene oxide 33.75 1603 1582 1.31 1.46
65. Viridiflorol 36.18 1612 1592 - -
66. Fokienol 37.29 1642 1596 - 0.10
67. Humulene epoxide ΙΙ 34.75 1631 1608 0.96 0.19
68. Murrola-4,10(14)-dien-1-pentol 38.58 1649 1630 - -
69. T-murrolol 38.08 1663 1644 - -
70. Eudesmol<β-> 38.38 1671 1649 - -
71. Cadinol<α-> 38.55 1676 1652 - -
72. Thujopsanone<3-> 38.89 1685 1653 - -
73. Bisabolol<α-> 37.71 1705 1685 - 0.05
74. Shyobunol 39.96 1713 1688 - -
75. Bergamotol<(Z)-α-trans-> 34.34 1564 1690 0.19 0.28
76. Mintsulphide 41.54 1759 1740 - 0.15
77. Bisabolenal<β-> 39.17 1485 1768 0.36 -
78. Hexadecanoic acid 49.12 1989 1959 - 0.03
% oil identified 95.83 84.08
Table 1: Comparative essential Oil Composition of Ocimum americanum Collected From Different Sites
05 1015 2025 3035 4045
Fresh Sun dried
Fig. 1: Comparison between essential oil composition of fresh and sun dried aerial parts of O.
americanum.
60 Acknowledgement
The authors are grateful to the Department of Science and Technology, New Delhi for financial support under DST INSPIRE fellowship, Dr. Chitra Pande, Professor, Department of Chemistry, D. S. B. Campus, Nainital for her sincere guidance, constant support and encouragement during the present research work and Head, Department of Chemistry, Kumaun University Nainital, for providing necessary laboratory facilities.
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