الفهرس 
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Abstract
SUMMARY
This study was conducted to evaluate the biological activity of plum seeds of Santarosa variety and olive seeds of Al-Agizi Al-Shami variety. where graduated polar extracts (hexane-ethanolic-aqueous) were prepared from both the seeds and the woody shell of the plants under study, where 12 extracts were obtained and placed in the freezer until use.
These extracts were used to study the following:
(1) The content of these plant parts of phenols, flavonides, tannins, sugars, amino acids and proteins.
(2) Its effect as antioxidant by estimating both:
(a) Reducing power.
(b) DPPH Radical Scavenging Activity.
(c) Superoxide Radical Anion (O2.-) Scavenging Activity.
(3) Its effect as anticancer on:
(a) HT-29 (Colorectal Cancer)
(b) MCF-7 (Breast Adenocarcinoma)
(4) Their effect as antimicrobial agents.
(a) Antibacterial activity on 5 strains
(Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Pectobacterium carotovorum, and Bacillus cereus)
(b) Activity as antifungal on Rhizopus stolonifer
(5) Separation and identification of some chemical components in extracts with the highest biological activity.
The results obtained are summarized in the following basic points:
1- The percentage of phenols, flavonides and tannins in each extract.
- In general, the ethanolic extract for each of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) compared to aqueous and hexane extract for each of the four plant parts.
- In particular, the ethanolic extract of olive seeds was the highest in phenols content (0.276%), the ethanolic extract of plum seeds was the highest in flavonoid content (0.191%) and the aqueous extract of olive seeds was the highest in tannin content (4.04%).
- The aqueous extract contains the highest percentage of sugars (6.07%) in PWS extract, amino acids (11.31%) in OS extract, proteins (1.43%) in PS extract.
2- Activity as antioxidant: -
(a) Reducing power
In general, there was a gradual and significant increase in the reduction power by increasing the concentration in all plant extracts under study.
-The ethanolic extract of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) showed the highest reducing strength compared to the aqueous and hexane extract of the same plant parts at the same concentrations used (0.5-15 mg/ml) because the ethanolic extract contains the highest percentages of phenols and flavonoids.
- The ethanolic extract of olive seeds showed the highest reducing power, possibly due to the content of olive seeds on the highest percentages of phenols and tannins compared to the rest of the four plant parts, at the same concentrations used (0.5-15 mg/ml).
(b) DPPH Radical Scavenging Activity
There is a gradual increase in scavenging activity with increasing concentration used in all plant extracts under study.
- The ethanolic extract of each plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) showed the highest scavenging activity compared to aqueous and hexane extract of the same plant parts at the same concentrations used (0.5-15 mg/ml).
- The ethanolic extract of olive seeds showed the highest scavenging activity over the rest of the four parts of the plant at the same concentrations used (0.5-15 mg/ml). This may be due to the content of olive seeds on the highest percentages of phenols and tannins compared to the rest of the other parts.
(c) Superoxide Radical Anion (O2.-) Scavenging Activity
- There is a gradual increase in the scavenging activity of the superoxide incision by increasing the concentration used in all plant extracts under study.
- The ethanolic extract of each of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) showed the highest scavenging activity compared to the aqueous and hexane extract of the same plant parts at the same concentrations used (1-5 mg/ml) and This may be due to the fact that the ethanolic extract contains the highest proportions of phenols and flavonoids.
- The ethanolic extract of plum seeds showed the highest scavenging activity over the rest of the four plant parts at the same concentrations used (1-5 mg/ml). This may be due to the content of plum seeds on the highest levels of flavonoids compared to the rest of the other parts
3- Activity as anticancer: -
(a) HT-29 (Colorectal Cancer)
- The results showed that all plant extracts (aqueous - ethanol - hexane) for each of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) have strong activity as anti-colon cancer (HT-29 cell line) and the ethanolic extract was the best followed by aqueous extract and then hexane at the same concentrations used (100-1200 μg/ml).
- The ethanolic extract of olive seeds also showed the highest anti-cancer activity, followed by plum seeds, followed by olive woody shells, then plum woody shells, and this may be due to the ethanolic extract containing the highest proportions of phenols and flavonoids, as well as due to the high activity of the ethanolic extract as an antioxidant.
(b) MCF-7 (Breast Adenocarcinoma)
- The results showed that the three extracts (aqueous - ethanolic - hexane) for each of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) have effectiveness as anti-breast cancer agents (MCF-7 cell line).
- The aqueous extract of (plum seeds – plum woody shells) had stronger anti-carcinogenic activity than the ethanolic and hexane extract of the same plant parts at the same concentrations used (100-1200 μg/ml).
- The ethanolic extract of (olive seeds – olive woody shells) had the best effect on carcinogenic cells than the aqueous and hexane extract of the same plant parts at the same concentrations used (100-1200 μg/ml).
- It was also found that the effect of these extracts was better if it worked as an anti-colon cancer than as an anti-breast cancer.
4- Activity as antimicrobial: -
(a) Antibacterial activity on 5 strains
(Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Pectobacterium carotovorum, and Bacillus cereus)
- First, for Pseudomonas aeruginosa, the ethanolic extract of the four plant parts showed the highest inhibitory activity of this microbe than the aqueous and hexane extract. The ethanolic extract of olive seeds was the best plant part in inhibiting microbial growth (26.83) at a concentration of 5 mg/ml.
- Second, for the microbe Escherichia coli, the ethanolic extract of the plant parts (plum seeds and plum woody shells) had a higher inhibition than aqueous and hexane extract. The aqueous extract of olive seeds was the best extract for inhibiting microbial growth at the same concentrations used (0.5-5 mg/ml).
- Third, for the Staphylococcus aureus microbe, the ethanolic extract of the plant parts (olive seeds and olive woody shells) is the best extract in inhibiting the growth of the microbe, while in the case of plum seeds and plum woody shells, the ethanolic extract is not given any inhibition.
- Fourth, for the microbe Pectobacterium carotovorum, the ethanolic extract of olive seeds was the best at inhibiting microbe growth and had the highest inhibition (16.33) at a concentration of 5 mg/ml.
- Fifth, for Bacillus cereus, the ethanolic extract of olive seeds was the best in inhibiting microbe growth and had the highest inhibition (15.83) at a concentration of 5 mg/ml.
- - from the previous results, it is clear that the high activity of the ethanolic extract of the plant parts may be due to their high content of bioactive compounds and these compounds may lead to the decomposition of the outer membrane of the bacteria under study.
(b) Activity as antifungal on Rhizopus stolonifer
The results showed that the three extracts (aqueous - ethanol - hexane) for each of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) did not show any inhibitory effect against the fungus Rhizopus stolonifer.
5- Separation and identification of some chemical components in extracts that have the highest biological activity.
- In this part, we explain the separation and identification of the basic chemical components for the most effective extracts of the plant parts under study, where:
(a) HPLC was used to identify the components of the ethanolic extract of the four plant parts (olive seeds – olive woody shell - plum seeds - plum woody shell) where the ethanolic extract was the most effective and the following examination was explained:
- First of all, for olive seed, they contained a high concentration of phenolic compounds, which were identified through HPLC and were as follows:
Ellagic acid (4075.71 µg/g DW) was the highest compound, followed by vanillin (2037.49 µg/g DW), syringic (1480.22 µg/g DW), chlorogenic (1024.78 µg/g DW), quercetin (935.56 µg/g DW), ferulic (837.58 µg/g DW), caffeic acid (833.34 µg/g DW, pyrocatechol (506.43 µg/g DW), daidzein (432.38 µg/g DW), gallic (283.35 µg/g DW), methyl gallate (225.49 µg/g DW), kaempferol (171.11 µg/g DW) and naringenin (102.10 µg/g DW, while Catechin and Hesperetin disappeared. On the other hand, the compounds rutin, coumaric, cinnamic and apigenin were present in low concentrations.
- Second, the woody shell of the olive seeds contains a high concentration of the following compounds: Ferulic acid (1910.37 μg/g DW), ellagic acid (811.24 μg/g DW), chlorogenic (381.44 μg/g DW) and gallic (120.31 μg/g DW). While Daidzein, Apigenin and Kaempferol disappeared. The rest of the other compounds were present in low concentrations.
- Third, plum seeds contain a high concentration of the following compounds: chlorogenic acid (797.48 µg/g DW), followed by ellagic acid (403.97 µg/g DW), gallic (253.79 µg/g DW) methyl gallate (195.92 µg/g DW), vanillin (173.85 µg/g DW), caffeic (125.04 µg/g DW), rutin (111.67 µg/g DW), catechin (106. 87 µg/g DW), syringic (106.16 µg/g DW) and quercetin (102.11 µg/g DW), while apigenin, kaempferol and hesperetin disappeared, while coumaric, ferulic, naringenin, daidzein and cinnamic acid were present in low concentrations.
- Fourth, for the woody shell of plum seeds, its extract was the least extract in its content of phenolic compounds that were separated by HPLC and were as follows: chlorogenic (219.92 μg/g DW), caffeic (206.25 μg/g DW), catechin (107.39 μg/g DW), while quercetin, apigenin and hesperetin disappeared, the rest of the compounds were present in low concentrations.
- In general, from the analysis of HPLC as well as the identification of chemical compounds in the ethanol fraction in the extracts under study, we note that the previous results are consistent with the content of phenols, flavonoids, high reducing power, high scavenging activity of free radical (DPPH) and scavenging activity on the Superoxide Anion (O2.-)
- Also, HPLC analysis confirms the strength of the anti-cancer activity of the ethanolic extract for the four plant parts and also shows the antibacterial activity of the extracts.
(b) GC-MS was used to identify the components of the hexane extract of (olive seeds –plum seeds). Total fatty acids in hexane extract from plum and olive seeds showed 9 fatty acids in each of them. Oleic acid (18:1 Cis 9), a monounsaturated fatty acid was identified as the major component in hexane extract which constituted 67.42% of the plum seeds, and 69.6% of the olive seeds. The olive seed extract had the highest amount of saturated palmitic acid (9%) as compared to the plum seed extract (5.53%). The polyunsaturated fatty acids (linoleic acid, 18:2 and linolenic acid, 18:3) content of olive seed was 16.23% and 0.28%. while linoleic acid and linolenic acid in plum seed were 24.46% and 0.05% respectively.
Recommendations
The plant extracts under study, especially the ethanolic extract, can be used as safe natural products such as antioxidants, anticancer and human pathogenic microbes.