Background
Methods
Formulation of plant crude extracts
Bacterial and fungal strains
Antimicrobial assay
Antioxidant assay
Anticancer assay
Cell culture
MTT assay
Qualitative phytochemical screening for secondary metabolites
Determination of total phenolic contents
Determination of total flavonoid contents
Isolation of active compounds
Identification of active compounds
Statistical analysis
Results
Antimicrobial activity
Microrganisms | Extracts | Zones of Inhibition (mm) | |||
---|---|---|---|---|---|
1.25 mg/mL | 2.5 mg/mL | 5 mg/mL | 10 mg/mL | ||
Acinetobacter junii
| Methanol | 0.0 ± 0.0 | 0.0 ± 0.0 | 7.5 ± 0.1** | 10 ± 0.2** |
Acetone | 0.0 ± 0.0 | 0.0 ± 0.0 | 7.9 ± 0.1** | 9.5 ± 0.1** | |
Ethyl acetate | 0.0 ± 0.0 | 8.0 ± 0.2** | 10.5 ± 0.2** | 11 ± 0.36** | |
Ethanol | 0.0 ± 0.0 | 0.0 ± 0.0 | 8.0 ± 0.36** | 10 ± 0.2** | |
Aqueous | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Serratia mercescens
| Methanol | 8.5 ± 0.2** | 8.8 ± 0.3** | 10.8 ± 0.26* | 11.6 ± 0.2** |
Acetone | 9.5 ± 0.36** | 9.7 ± 0.17** | 10.5 ± 0.60* | 12.8 ± 1** | |
Ethyl acetate | 10.5 ± 0.26* | 10.8 ± 0.1** | 11.6 ± 1.2* | 12.5 ± 1.1* | |
Ethanol | 9.0 ± 0.60** | 10.0 ± 0.17** | 12.1 ± .95* | 13.3 ± 0.51** | |
Aqueous | 7.3 ± 0.33** | 7.8 ± 0.57** | 9.0 ± 0.60** | 10.0 ± 0.20** | |
Enterobacter cloacae
| Methanol | 7.3 ± 0.26** | 7.5 ± 0.20** | 8.6 ± 0.36** | 10.6 ± 0.20** |
Acetone | 0.0 ± 0.0 | 0.0 ± 0.0 | 7.5 ± 0.3** | 8.5 ± 0.1** | |
Ethyl acetate | 7.8 ± 0.78* | 8.5 ± 0.1** | 10.5 ± 0.26** | 12.5 ± 0.50** | |
Ethanol | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Aqueous | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Pseudomonas aeruginosa
| Methanol | 7.5 ± 0.20** | 8.5 ± 0.1** | 10.0 ± 0.20** | 12.1 ± 0.43** |
Acetone | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Ethyl acetate | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Ethanol | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Aqueous | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Salmonella typhi
| Methanol | 7.8 ± 0.3** | 8.8 ± 0.52** | 10.8 ± 0.65** | 12.17 ± 0.43** |
Acetone | 7.1 ± 0.6* | 7.8 ± 0.30** | 8.2 ± 0.36** | 8.9 ± 0.43** | |
Ethyl acetate | 8.8 ± 0.28** | 9.3 ± 0.45** | 10.0 ± 0.20** | 12.5 ± 0.62** | |
Ethanol | 7.1 ± 0.26** | 8.0 ± 0.34** | 10.5 ± 0.50** | 11.8 ± 0.40** | |
Aqueous | 7.6 ± 0.40** | 8.8 ± 0.40** | 9.5 ± 0.43** | 10.0 ± 0.34** | |
Candida albicans
| Methanol | 8.5 ± 0.30** | 10.5 ± 0.78** | 11.5 ± 0.36** | 12.0 ± 0.8** |
Acetone | 10 ± 0.34** | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | |
Ethyl acetate | 9.0 ± 0.20** | 9.5 ± 0.51** | 10 ± 0.20** | 0.0 ± 0.0 | |
Ethanol | 10.0 ± 0.1** | 10.8 ± 0.40** | 11.0 ± 1.3* | 11.3 ± 0.55** | |
Aqueous | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 |
Controls | Chemicals | Zone of Inhibition (mm) | |||||
---|---|---|---|---|---|---|---|
A. junii
|
S. mercescens
|
E. cloacae
|
P. aeroginosa
|
S. typhi
|
C. albicans
| ||
Positive | Tigecycline Amikacin Cefepime | 12.1 ± 0.2 8 ± 0.20 0.0 ± 0.0 | 15.0 ± 0.50 13.3 ± 0.28 0.0 ± 0.0 | 16.1 ± 0.28 20.1 ± 0.28 19.8 ± 0.28 | 12.5 ± 0.50 12.0 ± 0.50 0.0 ± 0.0 | 16.0 ± 0.50 20.1 ± 0.29 17.0 ± 0.50 | 13.2 ± 0.2 0.0 ± 0.0 12.1 ± 0.3 |
Negative | DMSO(a) | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 |
Antioxidant assay
Test material | DPPH radical IC50 in μg/mL | |
---|---|---|
Plant extracts | Methanol | 2.49 ± 0.01* |
Ethanol | 2.41 ± 0.02* | |
Acetone | 3.06 ± 0.04** | |
Ethyl acetate | 2.74 ± 0.02* | |
Aqueous | 3.03 ± 0.04** | |
Standards | Gallic acid | 2.52 ± 0.02 |
Ascorbic acid | 2.54 ± 0.00 |
Anticancer assay
Cell Lines | Extracts | % Cell death | I.C50 | |||
---|---|---|---|---|---|---|
100 μg/μL | 150 μg/μL | 200 μg/μL | 250 μg/μL | μg/μL | ||
HCEC | Methanol | 2.3 ± 0.76 | 1.83 ± 0.76 | 3.43 ± 0.50* | 13.7 ± 2.42* | 21.09 |
Ethanol | 1.2 ± 0.52 | 6.23 ± 1.02* | 9.20 ± 0.81* | 12.8 ± 1.01* | 16.23 | |
Taxol | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 ± 0.0 | 0.0 | |
Tamoxifen | 14.5 ± 0.5 | 19.3 ± 0.7 | 26 ± 0.5 | 26.8 ± 0.7 | 6.29 | |
Hep2 | Ethanol | 6.06 ± 0.49** | 11.83 ± 0.65** | 21.0 ± 0.90* | 28.26 ± 0.25* | 7.28 |
Taxol | 26.5 ± 0.5 | 31.76 ± 0.66 | 35.53 ± 0.25 | 45.16 ± 0.60 | 3.97 | |
HeLa | Methanol | 16.7 ± 0.70** | 22.56 ± 0.40** | 27.73 ± 0.2** | 32.46 ± 0.61** | 5.45 |
Ethanol | 5.46 ± 0.40** | 15.53 ± 0.37** | 18.43 ± 0.4** | 22.9 ± .075** | 8.18 | |
Taxol | 11.46 ± 0.40 | 19.7 ± 0.51 | 24.63 ± 0.50 | 30.56 ± 0.45 | 6.07 | |
MCF-7 | Methanol | 23.33 ± 0.7** | 26.23 ± 0.25** | 42.4 ± 0.36** | 43.6 ± 0.45** | 4.0 |
Ethanol | 6.8 ± 0.52* | 11.5 ± 0.37** | 16.1 ± 0.1** | 23.56 ± 0.45** | 8.79 | |
Tamoxifen | 12.46 ± 0.40 | 17.36 ± 0.55 | 24.63 ± 0.50 | 28.56 ± 0.45 | 6.4 |
Phytochemical screening
EXTRACTS | Methanol | Ethanol | Acetone | Ethyl acetate | Aqueous |
---|---|---|---|---|---|
Steroids | + | + | + | + | + |
Alkaloids | + | + | - | + | - |
Tannins | + | + | - | + | + |
Phlobatannins | + | + | + | + | + |
Monoterpenes | + | + | + | + | + |
Flavonoids | + | + | + | + | + |
Coumarins | + | + | - | - | + |
Cardiac Glycosides | + | + | + | + | + |
Saponins | + | + | + | + | + |
Diterpenes | + | - | - | - | + |
Anthraquinones | + | + | - | - | + |
Phenols | + | + | + | + | + |
Total phenolic contents
S.No | Plant extracts/ chemicals (all 400 μg/mL) | Total phenolic contents (amg of GAE/g of extract) | Total flavonoid content (bmg QE/g dried extract) |
---|---|---|---|
1 | Methanol | 2.328 ± 0.003** | 5.00 ± 0.04** |
2 | Ethanol | 1.769 ± 0.001** | 3.73 ± 0.08** |
3 | Acetone | 1.535 ± 0.004** | 3.31 ± 0.06** |
4 | Ethyl acetate | 1.318 ± 0.002* | 3.70 ± 0.04** |
5 | Aqueous | 0.535 ± 0.002** | 1.29 ± 0.03** |