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IDENTIFICATION OF BIOACTIVE COMPOUNDS OF NYCTANTHES ARBORTRISTIS LINN BY GC-MS


Mrunal K. Shirsat*1, Indra Jeet Singhvi2, Khemchand Gupta3, Ayush Garg4
1. Dept. of Pharmacognosy, Pacific College of Pharmacy, Udaipur- Rajasthan, India
2.  Dept. of Pharmaceutical Chemistry, Pacific College of Pharmacy, Udaipur, Rajasthan – India
3.  Dept. of Pharmaceutics, Pacific College of Pharmacy, Udaipur, Rajasthan – India
4.  Dept. of Pharmaceutics, Pacific College of Pharmacy, Udaipur, Rajasthan – India

ABSTRACT

The objective of the present study is to determine possible bioactive compounds of the different extract fractions of entire plant of Nyctanthes arbortristis linn. The present research work was carried out by using GC-MS analysis, while mass spectra of the compounds found in the extract fraction was conducted by using the database of national institute of standard and technology (NIST) having more than 62000 patterns.

Forty components from different extract fraction of entire plant were identified. Active principles with their retention time, area, height of peak, percentage of area of peak were obtained. The research reveals the potential of entire plant of Nyctanthes arbortristis Linn as a good source of bioactive such as fatty acid esters, alcohol, hydrocarbon, aldehyde, ketones, amide, terpenes, sterols that justify used of this plant for its various ailments for traditional practitioners.

Keywords: GC-MS, Nyctanthes arbortristis linn, Phytol, Alpha-Comphenol, Extraction, Furfural.

INTRODUCTION

The flowers of Nyctanthes arbortristis linn was used as stomachic, carminative, astringent to bowel, antibilious, expectorant, hair tonic and in the treatment of piles and various skin diseases 1 and in the treatment of ophthalmic purposes 2. The bright orange corolla tubes of the flowers contain a coloring substance nyctanthin, which is identical with α- Crocetin (C20H24O4) from Saffron. The corolla tubes were formerly used for dyeing silk, sometimes together with Safflower or turmeric 3.

Traditionally the powdered stem bark is given in rheumatic joint pain, in treatment of malaria and also used as an expectorant 4. The bark is used for the treatment of snake bite and bronchitis [1, 5]. The resulting paste of Nyctanthes arbortristis on mixing with Arjuna bark is rubbed on the body to treat internal injury and for joint broken bones [3]. Medicinal used for the treatment of various diseases such as sciatica, chronic fever, rheumatism, and internal worm infections, and as a laxative, diaphoretic and diuretic 6. Leaves are used in cough. Leaf juice is mixed in honey and given thrice daily for the treatment of cough. Paste of leaves is given with honey for the treatment of fever, high blood pressure and diabetes 7. Therefore characterization extracts fraction of medicinal plant necessary due to the numerous benefits to the sciences and society.

MATERIAL AND METHODS

Plant Material

The plant Nyctanthes arbortristis linn     were collected from local area of Udaipur in the month of January 2011 and authenticated from Department of botany Nagpur University. Voucher specimen no. 9112 was deposited in university.

Chemicals

All solvents used in this present study such as hexane, Petroleum ether, chloroform and methanol were analytical grade.

Extraction

Nyctanthes arbortristis linn entire plant powders (500g) were extracted with petroleum ether, Chloroform methanol solvent (1000 ml, 72 h) by using successive hot Soxhlet extractor. After extraction, it was filtered and each solvent was evaporated by using rotary evaporator (Yamato Rotary Evaporator, Model RE 801). The solvent free Petroleum ether, Chloroform and methanol crude extract (11.5g, 10.6g, 9.6g respectively) was obtained that was free from particle and dried under vacuum then fractionation by different solvent by formation of three different faction i.e. Petroleum ether, Chloroform, Methanol then send by GC-MS Study.

Preliminary Phytochemical Screening and Fluorescence Analysis

The fresh plants were subjected to shade drying (22oC) for two weeks and then proceed at laboratory mail. Air dried coarsely powder of plant was 1 kg coarse powder was extracted with petroleum ether, chloroform, ethanol and distilled water in soxhlet extractor by continues successive hot method. Finally the extract was collected and concentrated for various qualitative phytochemical tests and fluorescence analysis in observed under UV 366 nm (Table 1) for identification of chemical constituents present in plant material and results were presented (Table 2, 3, 4).

GC-MS Analysis

The GC-MS analyses were carried out in Perkin Elmer, auto system XL GC+.

  • Carrier gas: helium with a flow rate of 0.7 ml/min.
  • Column temperature : 5minute in 180oC, 180-260oC at 3oC/min., 5minute in 260oC, 260-280 at 0.2oC, and finally 5 in 280oC injector temperature 280oC to 290oC detector temperature.
  • Volume injected: 1μL of sample
  • Ionization potential: 70 eV
  • Ion source temperature: 290oC

Identification of compound GC-MS chromatogram of Petroleum ether fraction, Chloroform fraction and methanol fraction of Nyctanthes arbortristis linn (Figure 1, 2, 3) showed peaks indicating the presence of 40 phyto-constituents. Interpretation of mass spectrum and GC-MS was conducted by using the database of national institute of standard and technology (NIST) having more than 62000 patterns (Figure 1, 2, 3, 4, 5, 6 and Table 2, 3, 4).

Identification of Compounds

The identity of the compounds in the extract was assigned by the comparison of their retention indices and Interpretation of mass spectrum and GC-MS was conducted by using the database of national institute of standard and technology (NIST) having more than 62000 patterns.

RESULTS AND DISCUSSION

The bioactive compounds present in the different extract fraction of entire plant of Nyctanthes arbortristis linn. The active principles with their retention time (RT), area, height, percentage of area and their structure were presented.

3.1: Study for petroleum ether Fraction of Nyctanthes arbortristis linn

Figure 1: GC-MS Chromatogram of Petroleum ether Fraction  

Table 1: Percentage extractive, fluorescence analysis, and preliminary phytochemical screening of entire plant of Nyctanthes arbortristis linn.

Solvent Extracts (%W/W) Fluorescence

Observed

(UV 366nm)

 

                         Chemical constituents
Alkaloids Sterols Terpenoid Sugar Glycoside Phenolic
Pet. Ether (60-800C) 2.75±0.076 Orange color —-  ++ +++ —–     —–
Chloroform 1.20±0.060 Reddish orange color —– ++ + ++ ++     ++
Ethanol 3.25±0.070 Yellowish Orange ++ ++ ++ + +++     ++
 Distilled Water 7.09±0.067 Greenish Blue —- —- + ++ ++      +

+ Present                                                                        +++ Prominently Present

++ Significantly Present                                                 —- Absent

 

Table 2: List of Probable Compound of Petroleum Ether Fraction for Peak at 37.26 RT (NAT)

Peak # Name of Compound RT (Retention time) Area Height Area %
1 Alpha-Comphenol 17.81 637.8 20,928 0.34
2 2-Methyl -4,6 quinolinediol 18.70 5,578.7 214,746 3.01
3 Octacosane 20.37 9,636.0 270,418 5.20
4 1-methyl-2-Chlorobenzen 20.61 3,131.5 123,348 1.69
5 Furfural 22.15 2,201.3 50,108 1.19
6 2,3,5-Trimethyl naphthalene 24.02 3,100.4 76,518 1.67
7 2,3-Dihydro benzofuran 25.27 729.6 18,522 0.39
8 Phytane 28.62 1,162.2 17,514 0.63
9 1,5-Heptadiene-3yne 33.17 6,226.1 56,904 3.36
10 1-Butoxy-2-ethyl-1-Hexene 34.91 21,454.6 214,531 11.57
11 Eicosamethyl- cyclodecasiloxane 37.26 57,276.4 495,939 30.90
12 1-Hexacosanol 41.33 4,808.5 34,068 2.59
13 Oxirane 43.03 5,807.4 36,893 3.13
14 2-Methylbenzoic acid 46.94 16,802.3 91,726 9.06

Figure 2: Phyto-components identified in the Petroleum ether fraction of the entire plant of Nyctanthes arbortristis linn by GC-MS

3.2. Study for Chloroform Fraction of Nyctanthes arbortritis linn.

Figure 3: GC-MS Chromatogram of Chloroform Fraction 

 

Table 3: List of Probable Compound of Chloroform Fraction for Peak at 37.22 RT (NAT)

Peak # Name of Compound RT (Retention time) Area Height Area %
1 Phytol 17.81 176.9 7,334 0.20
2 2-Methyl -4,6 quinolinediol 18.70 3,087.4 114,707 3.48
3 Octacosane 20.37 3,766.9 111,273 4.25
4 1-Methyl -2-Chlorobenzene 20.61 1,925.7 74,689 2.17
5 Furfural 22.15 1,284.5 28,377 1.45
6 2,3,5-Trimethyl naphthalene 24.02 2,195.7 48,267 2.47
7 Tetracotane 25.51 748.2 18,323 0.84
8 Alpha tocopherol 26.73 297.6 8,880 0.34
9 1,5-Heptadiene-3yne 33.17 2,116.7 26,779 2.39
10 Betulin 33.96 11,435.2 142,515 12.89
11 1-Butoxy-2-ethyl-1-Hexene 34.91 14,741.3 156,967 16.61
12 Tetradecane 37.22 18,846.5 186,492 21.24
13 2-Methylpropyl Phenol 41.35 3,687.8 25,493 4.16
14 Oxirane 43.03 4,241.6 29,012 4.78
15 2-Methylbenzoic acid 46.94 7,567.0 42,806 8.53

Figure 4: Phyto-components identified in the Chloroform fraction of the entire plant of Nyctanthes arbortristis linn by GC-MS  

3.3. Study for Methanol Fraction of Nyctanthes arbortritis linn.

Figure 5: GC-MS Chromatogram of Methanol Fraction

Table 4 List of Probable Compound of Methanol Fraction for Peak at 28.62 RT (NAT)

Peak # Name of Compound RT(Retention time) Area Height Area %
1 Alpha-Comphenol 17.81 727.3 17,812 0.72
2 2-Methyl -4,6 quinolinediol 20.37 2,090.5 79,137 2.07
3 Furfural 22.15 767.4 21,276 0.76
4 Squalene 23.76 1,716.9 45,729 1.70
5 Bis(2ethylhexyl) phthalate 25.29 2,593.6 74,022 2.57
6 Heneicosane 31.34 12,994.1 197,882 12.89
7 1-Butoxy-2-ethyl-1-Hexene 34.93 7,223.5 75,344 7.17
8 Tetradecane 37.24 15,571.3 147,885 15.45
9 D-erythro pentose,2-deoxy 41.35 2,101.6 11,818 2.09
10 Hexadecyl 43.05 2,263.2 15,822 2.25
11 11-Tridecane-1-ol 46.99 5,251.0 31,501 5.21

 

Figure 6: Phyto-components identified in the methanol fraction of the entire plant of Nyctanthes arbortristis linn by GC-MS

Conclusion

Among the identified Phytochemical of different fraction of Nyctanthes arbortritis linn: Petroleum ether, Chloroform and methanol fraction different type bioactive chemical constituents useful future study for nematicide, fungicide, anti-helminthes, anti-inflammatory, anti-diarrhea, ant-microbial, anti-oxidants, analgesics, antibacterial, anti-cancer, anticoronary, insectifuge, anti-androgenic, anti-arthritic all activity was observed in different database.

Acknowledgements

The authors are grateful to the Sicart (sponsored by Department of science & Technology, Govt. of India, New Delhi) for analysis of GC-MS.

REFERENCES

  • Wallander, E, and Albert, V. A, “Phylogeny and classification of Oleaceae based on RPS16 and TRNL-F sequence data,” American Journal of Botany, 2000; 87: 1827-1841.
  • Nawaz, Ahmed, Hossain, M, Karim, M, Khan, M, Jahan, R, and Rahmatullah, M, “An Ethnobotanicals Survey of Jessore district in Khulna Division Bangladesh,” American-Eurasian Journal of Sustainable Agriculture, 2009; 3: 238-243.
  • Nair, R, Kalariya, T, and Chanda, S, “Antibacterial activity of some selected Indian Medicinal Flora,” Turkish Journal of Biology, 2005; 29: 41-47.
  • Narendhirakannan, R. T, and Smeera, T, “In-vitro antioxidant studies on ethanolic extracts of leaves and stems of Nyctanthes arbortristis L. (Night-flowering jasmine),” International Journal of Biological and Medical Research, 2010; 1:188-192.
  • Kumar, S, Gupta, P, Sharma, S, and Kumar, D, “A review of immune-stimulatory plants,” Journal of Chinese Integrative Medicine, 2011; 9: 117-128.
  • Kannan, M, Singh, Ajay, Kumar, T. T. A, Jegatheswari, P, and Subburayalu, S, “Studies on immune-bioactivities of Nyctanthes arbortristis (Oleaceae),” African Journal of Microbiology Research, 2007; 1: 088-091.
  • Gadgoli, C, and Shelke, S, “Crocetin from the tubular calyx of Nyctanthes arbortristis,” Natural Product Research, 2010; 24: 1610-1615.