ANEXO IX: PROTOCOLOS DE TRASMISION DE DATOS DE LOS LIMITADORES
JUNTA DE ANDALUCIA
TABLE 10: DPPH radical scavenging activity of leaf essential oil, saponin and methanolic extract with ascorbic acid (standard)
Samples Concentration (μg /ml) Inhibition (%) IC50 (μg/ml)* 10 14.945 20 57.126 Ascorbic acid 30 95.627 19.07±0.02 40 96.572 50 97.653 10 9.845 20 45.812
Leaf essential oil 30 90.291 22.14±0.04
40 94.951 50 96.674 10 11.162 20 29.095 Saponin 30 91.810 23.79±0.01 40 95.154 50 95.863 10 9.845 20 46.826 Methanolic extract 30 91.000 22.00±0.01 40 94.343 50 95.560
Data are expressed as mean ±SD.*Values statistically different P > 0.05as measured by Tukey HSD test.
Fig. 3:Radicalsc avenging activity (%inhibition) and IC 50 values of methanolic extract, leaf essential oil ands aponin with ascorbic acid as stdin DPPH assay.
Fig.4:Antimicrobial activity in the methanolic extract, leaf essential oil ands aponin
1. Trichophytonrubrum ; 2. Cryp to coccusneo for mans; 3. Staphylococcusaureus4. Escherichiacoli
C-Positive Control ; a-leaf essential oil(neat);b- methanoli cextract ;c-saponin ;d- negative control(methanol)
TABLE 11: MIC values (mg/ml) for leaf essential oil, methanolic extract and saponin
Organism Leaf essential oil Methanolic extract Saponin
Trichophyton rubrum 0.12 0.25 00
Cryptococcus neoformans 0.25 0.50 00
Staphylococcus aureus 0.25 0.50 1.00
TABLE 12: Screening of antimicrobial activity in the leaf essential oil, methanolic extract and saponin
Organism Zoneofinhibition(mm) (mean±SD)
Leafessential Methanolic Saponin
oil extract Staphylococcus aureus 16.33±1.15 12.66±1.15 11.00±0.00 Staphylococcus epidermidis 7.13±0.11 - - Salmonella typhi - - - Trichophytonrubrum 17.33±0.57 16.66±0.56 - Cryptococcus neoformans 11.00±0.00 11.33±0.57 - Candida albicans 8.67±0.57 8.00±0.00 - Enterococcus faecalis - - - Pseudomonas aeruginosa 7.09±0.57 6.80±0.00 - Klebsiella pneumoniae - - -
Fig. 5: MIC values (mg/ml) for leaf essential oil, methanolic extract and saponin.
TABLE 13: In vitro cytotoxicity of leaf essential oil , methanolic extract and saponin against HeLa, A-549 and PC-3 cell lines
Cellline Concentration Cytotoxicity (%)
(µg/ml) Leafessential oil Methanolic Saponin
extract HeLa 1000 74.03±1.2 76.60±1.0 52.77±1.0 500 69.28±1.5 70.39±1.0 45.77±1.2 250 60.14±4.3 59.22±1.3 40.77±1.8 125 35.65±1.4 47.94±1.3 36.38±0.8 62.5 26.62±1.1 23.25±2.0 25.75±1.2 31.25 12.43±1.2 NA 25.33±1.3 A-549 1000 80.63±0.5 90.41±2.3 78.63±0.5 500 59.09±1.0 75.91±2.1 69.09±1.0 250 38.96±1.4 73.22±1.1 48.96±1.1 125 31.88±1.2 63.69±1.6 33.88±1.2 62.5 21.33±2.2 31.05±2.4 22.33±2.3 31.25 NA 20.02±1.3 21.88±1.4 PC-3 1000 89.77±0.1 76.70±1.2 31.25±0.6 500 86.24±1.1 65.23±2.3 21.91±1.0 250 86.09±0.8 36.61±1.4 13.91±0.9 125 69.00±0.9 18.73±3.4 9.59±1.0 62.5 60.11±0.6 9.69±1.00 NA 31.25 52.08±1.1 NA NA Data expressed as mean ± SD
Fig. 6:Invitrocytotoxicity ofleafessentialoil,methanolicextractand saponin against HeLa, A-549 and PC-3 celllinesrespectively
[LE-leafessentialoil;ME-Methanolic extract;SP-Saponin]
TABLE 14 : CTC 50 values of leaf essential oil, methanolic extract ans saponin against various cell lines
Sample CTC50 (µg/ml)
HeLa A-549 PC-3
Leafessential oil 203.33±1.9a 393.33±1.3b
42.5±0.08c
Methanolic extract 340.00±1.4a 103.33±1.9b
a
Data are expressed as mean ± SD. *Values statistically differat P >0.05 with in the row as measured by
Tukey HSD test.
Fig.6:In vitrocytotoxic activity of the leaf essentialoil, saponin and methanolicextract against HeL a, A-549 andPC-3celllines in terms of CTC50values
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