The document compares the phytochemical profiles and biological activities of green and black Ocimum sanctum (tulsi). Key findings include: - Black tulsi has higher total phenolic and eugenol content than green tulsi, while green tulsi has more volatile oil. - Black tulsi showed stronger antioxidant activity and broader-spectrum antimicrobial effects against various bacteria compared to green tulsi. - The higher biological activity of black tulsi correlates with its higher phenolic and eugenol content levels. - The study supports the belief that black tulsi has greater medicinal value than green tulsi.

1. PHYTOCHEMICALAND BIOLOGICAL
EVALUATION OF GREEN AND BLACK
OCIMUM SANCTUM
Shree Joshi, Anshu Srivastava, Milind Bagul, Avani Kothari and Neeta Shrivastava
B. V. Patel Pharmaceutical Education and Research Development (PERD) Centre,
Thaltej, Ahmedabad – 380 054
Phone: 079/ 27439375, Website: www.perdcentre.com

2. THE PLANT
Ocimum sanctum (Lamiaceae)
Chemical constituents
Major: Volatile oil (consists eugenol - 70.5%, nerol,
caryophyllene camphene, etc.)
Minor: Alkaloids, glycosides, saponins and tannins
Therapeutic uses
Classical: Indigestion, cough, malarial fever, wound, poisoning,
eye diseases, etc.
Scientific: Antitubercular, antiulcerogenic, antifertility,
anticancer, antiasthmatic, hepatoprotective, immunomodulatory,
etc.

3. Green Tulsi (GT) Black Tulsi (BT)

4. OBJECTIVE
Comparative evaluation of phytochemical profile
and biological activity of Green Tulsi and
Black Tulsi

5. PHYTOCHEMICAL EVALUATION

6. Parameters
ESTIMATION OF TOTAL PHENOLICS
(Singleton and Rossi method, 1965)
ESTIMATION OF VOLATILE OIL
(Hydrodistillation method)
ESTIMATION OF MARKER COMPOUND
EUGENOL
(HPTLC method)

7. Parameters Green Tulsi Black Tulsi
Total phenolic
content
0.65% 1.75%
Volatile oil 2.0% 1.6%
Eugenol 0.4% 0.6%
Results

8. Chromatogram for Estimation of Eugenol
Mobile phase - Toluene: Ethyl Acetate: Formic Acid (6: 4: 0.4)
Eugenol
BT
GT

9. TLC FINGERPRINTING
1) Test sample
Volatile oil and Methanolic extract
2) Mobile phase
Volatile oil with Eugenol
Toluene: Ethyl Acetate (8.5: 1.5)
Methanolic extract
Toluene: Ethyl Acetate: Formic acid (6: 4: 0.4)

10. Fingerprinting of Volatile Oil with Eugenol
Rf VALUE OF BANDS
GT STANDARD (EUGENOL) BT
0.56 - -
0.67 0.67 0.67
0.77 - -
0.97 - 0.97
BT
EUGENOL
GT

11. Fingerprinting of Methanolic Extract
TLC Fingerprinting after derivatization
GT BT
Color of bands Rf value
GT BT
Violet 0.07 0.07
Violet 0.12 0.12
Light blue 0.16 0.16
Light blue 0.33 0.33
Dark blue 0.52 0.52
Dark green 0.59 0.59
Purple 0.70 0.70
Dark green 0.73 0.73
Pink 0.81 0.81
Violet 0.95 0.95
Violet 0.98 0.98

12. BIOLOGICAL EVALUATION

13. FREE RADICAL SCAVENGING ACTIVITY
DPPH MODEL
1) Test sample – Methanolic extract
2) Stock solution - DPPH in methanol
3) Reaction mixture
2 ml 100 l extract in different
+ +
methanol DPPH concentrations
Incubation for
15 mins.
Absorbance recorded at 516 nm

14. Concentration of
extract (g)
% Inhibition
GT BT
50 4.6 20.2
100 7.8 46.6
200 17.9 80.7
300 22.7 83.1
400 30.7 86.7
Results

15. ANTIMICROBIAL ACTIVITY
1) Test sample
Volatile oil and petroleum ether extract
2) Microorganisms
Eleven bacterial ATCC cultures
3) Medium
Antibiotic assay medium No. 1
4) Positive control
Ciprofloxacin: 4 g/ ml
AGAR PLATE METHOD

16. Bacterial cultures (ATCC No.) Blank Positive
control
Volatile oil Petroleum ether extract
GT
(2l/ ml)
BT
(2l/ ml)
GT
(3 mg/ ml)
BT
(3 mg/ ml)
Bacillus cereus (11778) +++ - - - +++ +++
Bacillus pumilis (14884) +++ - - - +++ +++
Bacillus subtilis (6633) +++ - - - +++ -
Bordetell bronchiseptia (4617) +++ - - - +++ -
Micrococcus luteus (9341) +++ - - - +++ -
Staphylococcus aureus (29737) +++ - - - +++ -
Staphylococcus epidermis (6538) +++ - - - +++ -
Escherichia coli ( 10536) +++ - + - +++ -
Klebsiella pneumonia (10031) +++ - - - +++ -
Pseudomonas aeruginosa (9027) +++ - ++ - +++ -
Steptococcus faecalis (8043) +++ - - - +++ -
Antimicrobial activity of Volatile oil and Petroleum ether extract with agar plate method.

17. Antimicrobial Activity of Volatile Oil
Blank Green Tulsi
Black Tulsi

18. Antimicrobial Activity of Petroleum Ether Extract of
Black Tulsi

19. SUMMARY
● Phytochemical profile and Biological activity
of Green and Black Tulsi are considerably
different
● Volatile oil content is more in Green Tulsi
while total phenolic content and eugenol
content is higher in Black Tulsi
● TLC fingerprinting profile of Volatile oil of
both the types of Tulsi showed variation
● Biological activity showed higher potential in
Black Tulsi

20. CONCLUSION
Black Tulsi showed higher potential in Biological
activity than Green Tulsi that may be attributed to
the High Eugenol and Total phenolic content
The study scientifically supports the belief that Black Tulsi is
medicinally more valuable than Green Tulsi