INTRODUCTION:
Demands of traditional herbal medicines are increasing day by day not only by the developing countries but also by the developed contries throughout the world. The demand is due to the increased acceptance of Ayurveda and traditional herbal medicines, because of having their no side effects, and such modern people relies more on drug resource of plant origin1.
Medicinal plant is used World Wide in management of healthcare problems since time immermorial and approximately 60-80% of the world’s population still depending on the tradicinal medicine. Currently, the global demand of herbal medicine is incrasing rapidly because of their safety margin and low cost. With the help of medicinal plants still many of the disease are getting cured.
An applicability of theis herb is wide with the decoction of the root being used to control food poisoing, reduce fever, as a lung tonic and a bronchodilator.
Leaves of Millingtonia hortensis are used as antipyretic, remedy for asthma, in sinusitis, as a cholagogue and tonic in folklore medicine. The dried flower are used as a treatment for astuma2,3. The flower were reported to contain hispidulin (6 methoxy -5,7,4’- trihydroxyflavone), scutellarein, and scutellarein-5-glucuronide, respectively. The flower also contain a flavonoids, crisimaritin (hortensin; 5,4’- dihydroxyflavone)4.
MATERIALS AND METHODS:
Collection of plant material:
The flowers of Millingtonia hortensis were collected from its natural habitat in and around Mannargudi, Thiuvarur district, Tamilnadu, India.
Preparation of flower extract:
Fresh plant was shade dried at room temperature for 10 days and powdered course led in to cherry using electric blender. The plant powder (10 gm) was taken and mixed with ethanol (250 ml) .The mixture was boiled until and it was reduced to one third. The extract was filtered with a muslin cloth. The filtrate was transferred in to china dish and was allowed to evaporate using water bath. The obtained paste form of the extract was used for phytochemical and antioxidant activity.
Preliminary phytochemical screening:
Phytochemical analysis of the extract was conducted, by the standard procedure5. By this analysis, the presence of several phytochemical like alkaloids, flavonoids, tannins, saponins, esters, resins, sugars and glycosides were tested.
Gas chromatography- Mass spectrometry Analysis:
For quantization (area%), the GC analyses were carried
out by using JEOL JMS-700 by the electron impact method where an electronic
accelerating voltage of 75eV and an ion accelerating voltage of 8-10 kv. The
reservoir inlet systems were used. The capillary columns were: non polar column
DS-5MS (J&W Scientific; 30m 
0.25 mm, film thicknes 0.25 
column TC-Wax (60 m × 0.25 mm, film
thickness 0.25µm). The dynamic range for the peak intensities was 3 digits, and
the accuracy of the mass number was 0.5. The oven temperature was programmed
from 40°-240° C at a rate of 4°C/ min and held at 240° C for 5 min. The injector
and detector temperatures were 240° C and 280° C. The flow rates of carrier gas
(He) were 1.8mL. GLC data reported are given as area percentage. He at 49.9 KP
a was used as carrier gas and the FID detector was maintained at 250° C. The
phyto constituents were identified on the basis of their retention data and by
using GC /MS analytical conditions. The mass spectra were recorded on a mass
spectrometer coupled to a JEOL JMS -700 GAS chromatograph (EI mode70eV, source
temperature 230° C, scanned mass ranged 35-350 amu). The characteristic
fragmentation patternshave been analyzed and compared to those of Wiley 275.L
database.
Identification of compounds:
The identification of the compound was based on comparison with the library spectra (NIST-1, NIS NIST-2, Wiley 275 and Adams libraries) of their relative retention indices with literature values6. The relative percentage amount of each component was calculated by comparing its average peak was calclulated by comparing its average peak area to the total areas. The name, molecular wight, molecular formula and structure of the components of the material were determined and the data are presented.
RESULTS AND DISCUSSION:
Preliminary Phytochemical Screening:
The results of the preliminary phytochemical analysis of flower extract of ethanol showed positive test for flavonoids, glycosides, carbohydrate, phenolic compound and reducing sugar and negative test for alkaloids, saponins, tannins, steroids, protein & aminoacid (Table 1).
Table 1: Preliminary phytochemical screening of ethanolic extract of Millingtoniahortensis
|
S. No |
Phyto Constituents |
Results |
|
1 |
Alkaloids |
_ |
|
2 |
Flavonoids |
+ |
|
3 |
Saponins |
_ |
|
4 |
Tannins |
_ |
|
5 |
Glycosides |
+ |
|
6 |
Steroids |
_ |
|
7 |
Phenolic compounds |
+ |
|
8 |
Carbohydrate |
+ |
|
9 |
Protein & aminoacid |
_ |
|
10 |
Reducing sugar |
+ |
+ indicates presence whereas – indicates absence
Prelimiary phytochemical screening revealed the presence of flavonoids, alkaloids, tannins, phenols and alkaloids are very important plant constituents because of their free radical scavenging ability7.
Gas Chromatography Mass Spectrophotometry Analysis (GC- MS):
GC–MS is one of the best techniques to identify the constituents of volatile matter, long chain, branched chain hydrocarbons, alcohols acids and of thirty five compounds (Phytochemical constituents) that could contribute the medicinal quality of the plant. The Phytochemical compounds were confirmed based on the peak area, retention time and molecular formula. The result revealed that Aspidoseprmidin 17-ol, 1- acety-19, 21-epoxy-15, 16-dimethoxy (R.T:27.90); 2-methyltetracosane (R.T:28.24); Alfaxalone (R.T: 29.17); cis-13, 16-Docasdienoic acid (R.T: 30.86); Olean-12-ene-3, 15, 16, 21, 22, 28-hexol (R.T: 33.14); Stigmasta-5, 22-dien-3-ol, acetate, (3β) (R.T: 34.20); β-Sitosterol (R.T: 35.72); Stigmasta-5,24(28)-dien-3-ol,(3β) (R.T:36.10); Betulin(R.T:37.86), were founds as along with other major components in the ethanol extract and the other minor components such as E-N-Formyl-L-lysine (RT:0.64); 4H-Pyran-4-one,2,3-dihydroxy-6-(RT:4.08); Benzoic acid(RT:4.46); 5-Hydroxymethlfurfural (RT:5.37); 4-Hydroxyhistamine (RT:8.40) (Table 2 & Figure 1)
The GC- MS analysis of Cassia italic leaves revealed the presence of seventeen compounds. The identified compounds possess many biological properties. For instance, 9,12,15-Octadecatrienoic acid, (Z,Z,Z)- Linolic acid (R/T 20.06) possesses anti-inflamatory, insectifuge, hypocholesterolemic, cancer preventive, nematicide, hepatoprotective, antihistaminic, antieczmic, antiacne, 5-alpha reductase inhibitor, antiandrogenic, antiarthiritic and anticoronary properties. N-Hexadecanioc acid- palmatic acid (R/T 17.25) can be an antioxidant, hypocholesterlomic, nematicide, pesticide, lubricant activities and hemolytic 5- alpha is a reductase inhibitors. Phytol- Diterpene (R/T 19.67) is an antimicrobial, anticancer, anti-inflammatory and diuretic agent8. 9,12,15- Octadecatrienoic acid, methyl ester, 9(Z,Z,Z)-n-Hexadecanoic acid, 1, 2–Benzendicarboxylic acid anddi-isooctyl ester were present in Caesalpinia sappan ethanol extract9.