PRELIMINARY PHYTOCHEMICAL SCREENING

AND ADAPTOGENIC ACTIVITY OF

Chlorophytum borivillianum ROOTS

Key Words: Chlorophytum borivillianum, saponins, alcoholic extract, adaptogenic activity

 ABSTRACT
BACKGROUND AND OBJECTIVE
In India, Chlorophytum borivillianum Linn. (Safed Musli) is under commercial cultivation for its peerless healing
and health-giving properties. Due to the wide range of applications, it is witnessing steadily growing use in Pharmaceutical, Phyto-
Pharmaceutical and Nutraceutical products. In Ayurveda, Safed Musli is reputed for its tonic and aphrodisiac properties. Since very
less phytochemical data is available for such an important herb, it is planned to carry out phytochemical screening. Also there is no
proper scientific evidence to show its adaptogenic activity hence it is selected to prove scientifically its adaptogenic activity.

METHOD
A. Phytochemical Investigation:
 Extraction - with alcohol & water.
 Successive extraction - with petroleum ether, butanol, ethyl acetate and alcohol.
 Qualitative chemical analysis.
 Thin Layer Chromatography.
 Isolation of Saponins.

B. Adaptogenic Activity:
Alcoholic extract & butanolic fraction of alcoholic extract of powdered roots were tested for adaptogenic activity
using cold stress method.

RESULTS AND CONCLUSION

Qualitative chemical analysis of various extracts showed presence of Carbohydrates, Proteins, Saponins & Steroids.
Two saponins (S1 & S2) were isolated using preparative TLC. Alcoholic extract showed significant adaptogenic activity.
SAFED MUSLI

Synonyms:
Telugu - Swetha Musli
Hindi - Khiruva
Sanskrit - Swetha Musli
Marathi -Safed Musli

Biological Source:
It consists of dried peeled tuberous roots of plant known as Chlorophytum borivillianum Linn. belonging to family Liliaceae.

Chemical Constituents:
Carbohydrates (42%), Proteins (8.9%), Steroidal Saponins (0.1 – 1.7%), Alkaloids (25%), Fibers (34%), Vitamins and
Polysaccharides.

Therapeutic Uses:
• Finds therapeutic application in Ayurveda, Unani & Allopathy, as an Adaptogenic.
• As a curative for physical weakness and many other illnesses.
• As a curative for natal and post-natal problems.
• As a general sex tonic.
• As an effective alternative to Viagra.
• As an immunity improving agent.
• As a remedy for Diabetes & Arthritis.
A.COLLECTION AND AUTHENTICATION

Peeled & dried roots of Safed Musli were collected from Dharme Aromatic And Medicinal Estates, Dist. Akola,
Maharashtra and was authenticated by Dr. U. S. Choudhary, Professor and head of the department of Botany, Amravati University,
Amravati, India.

B.PHYTOCHEMICAL INVESTIGATIONS
1.Extraction
The dried powdered roots of Safed Musli were subjected to systematic phytochemical investigation by successively
extracting with various organic solvents of increasing polarity and the respective extracts were weighed and percentage extractive
values were determined.

% DRY WEIGHT
EXTRACTS COLOUR ODOUR CONSISTENCY
IN Gm.
Alcoholic 7.50 Dark brown Characteristic Sticky
Aqueous 54.06 Dark brown Characteristic Sticky
S U C C E S I V E E X T R A C T I O N
Petroleum ether
0.21 Pale yellow Characteristic Waxy
(40-600C)
Butanol 2.50 Dark brown Characteristic Sticky
Ethyl acetate 0.15 Brown Characteristic Sticky
Alcohol 3.33 Dark brown Characteristic Sticky
Water 45.69 Dark brown Characteristic Sticky
2. Qualitative Chemical Investigation
The qualitative chemical investigations were carried out to check the presence of various phytoconstituents. The
tests revealed the presence of Carbohydrates, Proteins, Steroids and Saponins.

Phytoconstituents Alcoholic Aqueous Successive extraction

extract extract
PE BU EA AL AQ

Carbohydrates + ve + ve - ve +ve +ve +ve +ve

Flavonoids - ve - ve - ve - ve - ve - ve - ve
Alkaloids - ve - ve - ve - ve - ve - ve - ve
Steroids + ve - ve + ve - ve - ve - ve - ve
Triterpenoids - ve - ve - ve - ve - ve - ve - ve
Tannins - ve - ve - ve - ve - ve - ve - ve
Proteins & AA’s + ve + ve - ve + ve + ve + ve + ve
Saponins + ve + ve - ve + ve + ve + ve + ve
KEY WORDS:
PE = Petroleum ether extract (40-600C) - = Absent
BU = Butanolic extract + = Present
EA = Ethyl acetate extract
AL = Alcoholic extract
AQ = Aqueous extract (by maceration)
3. Thin Layer Chromatography (TLC )

 TLC was performed to confirm the presence of saponins.

 TLC Profile For Saponins
 Stationary phase - Silica Gel G
 Mobile phase - Toluene: Chloroform: Acetone: Methanol ( 50 : 20 : 08 : 05 )
 Visualizing agent - Carr-Price reagent.
 Colour of the spots - Pink

Extracts No. of spots Rf Value

Alcoholic 2 spots 0.32

0.12
Aqueous 2 spots 0.32
0.12
Successive Butanolic 2 spots 0.32
0.12
Successive Ethyl Acetate 2 spots 0.32
0.12
Successive Alcoholic 2 spots 0.32
0.12
4. ISOLATION OF SAPONINS
I. Isolation Of Crude Saponins
Defatting with petroleum ether by hot extraction.
Exhaustive extraction with methanol.
Methanolic extract - conc. get a semisolid residue, suspended in water.
Fractionated with n-butanol in a separating funnel, combined
and concentrated to little volume.
Ethyl acetate was added drop-by-drop to precipitate saponins.

Wt. of powdered drug % Yield of crude

taken saponins obtained
70.00 gm. 0.44% w/w

II. Isolation Of Pure Saponins

From the crude saponin mixture obtained from the above method, pure saponins (S1 & S2) were isolated by Preparative TLC.

Preparative TLC Profile For Crude Saponins

Fraction Solvent system No. of Rf value Yield

Bands
Crude Saponins Toluene: Chloroform: 0.32 (S1) 20 mg
obtained from Acetone: Methanol 2
0.12 (S2) 15 mg
alcoholic extract. (50:20:08:05)
C. ADAPTOGENIC ACTIVITY

I. Materials And Methods

1. Animal selection:
Wistar strain Albino rats of either sex weighing between 130 and 180gm were selected for activity. Albino mice of either sex
weighing between 20 and 30gm were selected for acute toxicity studies.

2. Extracts Used:
Alcoholic extract and butanolic fraction of alcoholic extract.
1% w/v CMC in water was used as a vehicle for both the extracts.

3. Acute Toxicity Study:

The Up and Down Staircase method was adopted, and accordingly doses of alcoholic and butanolic fraction of alcoholic
extracts were calculated (500 mg/kg b.w).

4. Adaptogenic Activity:
Model Used: Cold stress method.
Procedure: The rats were grouped into four groups each containing 8 rats.
Group I --------- Normal
Group II -------- Stress induced
Group III------- Stress + Alcoholic Extract
Group IV------- Stress + Butanolic fraction of Alcoholic Extract
 Stress was induced by exposing the rats to cold environment (4 C  1C) for 3 hr under immobilized state
in a plastic cage. Adaptogenic activity of Safed Musli was evaluated by administrating the alcoholic extract and butanolic
fraction of alcoholic extract in a dose of 500mg/kg orally 1 hour prior to stress stimulus. Results were analyzed using
student “t” test.

ADAPTOGENIC ACTIVITY OF VARIOUS EXTRACTS OF SAFED MUSLI

Adrenal Weight Spleenic Weight BUN

Groups Ulcer Index (mg/100g) (mg/100g) (mg%)
I 0.375 14.218 395.235 21.128
(Normal)  0.125  0.232  8.011  0.053
II 4.625 22.803 424.625 28.00
(Stress Induced)  0.125  1.289  9.051  1.20
III 1.500** 16.368** 402.648 26.520
(Stress + Alco. Ext.)  0.125  0.918  17.638  0.965
IV 4.875 15.293** 410.363 25.370
(Stress +BFAE)  0.125  1.824  9.561  1.219.
P-value < 0.0001 < 0.0034 < 0.4188 < 0.2884

( BFAE - Butanolic fraction of alcoholic extract ; BUN – Blood urea nitrogen )

Results are expressed as Mean  SEM. (n = 8), ** p < 0.01
Group II compared with Group III & Group IV.
 CONCLUSION

 Qualitative chemical analysis of various extracts showed presence of Carbohydrates, Proteins,

Saponins & Steroids.

 The results obtained led to the conclusion that the two isolated compounds ( S1 & S2 ) are

saponins in nature.

 The results also revealed that alcoholic extract ( 500 mg/kg. b.w. ) showed moderate significant

adaptogenic activity whereas the butanolic fraction of alcoholic extract showed a greater

activity.

 Both the saponins identified ( S1 & S2 ) may be further investigated for its structural

conformity.

conformity.
REFERENCES

1. Kokate C.K, Purohit A.P, Gokhale S.B., Textbook of Pharmacognosy. 17th ed. New Delhi: CBS Publishers &
Distributors; 2001.
2. Singh A, Khanuja S.P.S, Singh S., Agronomic practices for the cultivation of Safed Musli in India. Natural Product
Radiance. 2003; 2 (6): 308 -13.
3. Gondi M, Shriniwasan K., Safed Musli – Herbal Viagra. Agrobios Newsletter.2004; 2 (11): 17-18.
4. http://www.safedmusli.net/applications.htm 06.06.2005
5. Gondi M, Shriniwasan K., Safed Musli – Herbal Viagra. Agrobios Newsletter.2004; 2 (11): 17-18.
6. Stahl E., Thin Layer Chromatography; 2nd ed. Heidelberg: Spinger-Verlag Berlin; 2003. 346-347 pp.
7. Mukherjee P.K., Quality Control of Herbal Drugs.1st ed. New Delhi: Business Horizons; 2002. 411 pp.
8. Kasture A.V, Wadodkar S.G, Mahadik K.R, More H.N., Pharmaceutical Analysis (Instrumental methods.) 2nd ed.
Pune: Nirali Prakashan; 1997.25-39 pp.
9. Ghosh M., Fundamentals of Experimental Pharmacology.1st ed. Kolkata: Scientific book Agency; 1984. 153-157 pp
10. Dadkar V.N, Joshi A.G, Jaguste V.S, Billimoria F.R, Dhar H.L., Antistress activity of Oscimum sanctum. (Tulsi).
Indian Drugs.1987; 25 (5): 172-75.
11. Kulkarni S.K., Handbook of experimental pharmacology. New Delhi: Vallabh Prakashan; 1999. 79 pp.
Document By
SANTOSH BHARADWAJ REDDY
Email: help@matlabcodes.com
1.Engineeringpapers.blogspot.com
2.www.matlabcodes.com
3.microcontroller-project-codes.blogspot.com
4.microcontroller-library.blogspot.com
5.arduino-projects-here.blogspot.com
6.labview-projects.blogspot.com
7.java-basics.blogspot.com
8.itsnanoworld.blogspot.com
More Papers,Projects and Presentations available on above sites.