Formulation of Ethyl Cellulose Microparticles Incorporated Pheophytin A Isolated from Suaeda vermiculata for Antioxidant and Cytotoxic Activities

Abstract

Background: This study is designed to discover a method for delivering an efficient potent pheophytin a (pheo-a) into more absorbed and small polymeric ethyl cellulose (EC) microparticles. Methods: Silica gel and Sephadex LH-20 columns were used to isolate pheo-a from the chloroform extract of the edible plant, Suaeda vermiculata. Pheo-a was incorporated into EC microparticles using emulsion-solvent techniques. The antioxidant activity of pheo-a microparticles was confirmed by the level of superoxide radical (SOD), nitric oxide (NO), and reducing power (RP) methods. Meanwhile, the cytotoxic effect of the product was investigated on MCF-7 cells using MTT assay. Results: Pheo-a was isolated from S. vermiculata in a 12% concentration of the total chloroform extract. The structures were confirmed by NMR and IR spectroscopic analysis. The formulated microparticles were uniform, completely dispersed in the aqueous media, compatible as ingredients, and had a mean diameter of 139 ± 1.56 µm as measured by a particle size analyzer. Pheo-a demonstrated a valuable antioxidant activity when compared with ascorbic acid. The IC50 values of pheo-a microparticles were 200.5 and 137.7 µg/mL for SOD, and NO respectively. The reducing power of pheo-a microparticles was more potent than ascorbic acid and had a 4.2 µg/mL for IC50 value. Pheo-a microparticles did not show notable cytotoxicity on the MCF-7 cell line (IC50 = 35.9 µg/mL) compared with doxorubicin (IC50 = 3.2 µg/mL). Conclusions: the results showed that water-soluble pheo-a microparticles were prepared with a valuable antioxidant activity in a wide range of concentrations with a noteworthy cytotoxic effect.

Suaeda vermiculata Aqueous-Ethanolic Extract-Based Mitigation of CCl4-Induced Hepatotoxicity in Rats, and HepG-2 and HepG-2/ADR Cell-Lines-Based Cytotoxicity Evaluations

“…For instance, the quercetin (12.45%), and quercetin-3- O -rutinoside (7.80%) were previously isolated from Suaeda japonica, Suaeda salsa, and Suaeda physophora [ 43 ], while kaempferol and kaempferol glycoside are also common constituents of the genus Suaeda plants, and have been reported both from Suaeda japonica and Suaeda asparagoides [ 43 ]. The LC-MS analysis of the aq.-ethanolic extract showed a base peak at m/z 872.1408 [M + H] + which was identified as the pheophytin-a (23.69%), the reported major chlorophyll-based constituent from Suaeda vermiculata species [ 48 ]. The fragmentation pattern for pheophytin-a showed mass peaks at m/z 256.26268 [phytyl-C 2 H 5 + H], 280.26241 [phytyl moiety], 593.15607 [M-phytyl], 577.12459 [M-phytyl–methyl], and m/z 615.13623 [M-phytyl + Na], which together confirmed the structure of the compound.…”

“…In the current study, the relatively high mg per gram content values of the phenolics and flavonoids are in the chloroform and ethyl acetate fractions compared to the mother liquor and confirms the higher antioxidant activity of these fractions over the mother liquor. Furthermore, the pheophytin-a, which was reported for the mild antioxidant activity, and isolated previously form Suaeda vermiculata chloroform fraction [ 48 ] as part of the ongoing study, also played a role in the antioxidant activity of the chloroform fraction. Additionally, the steroidal constituent, β-sitosterol [ 48 ], along with other unsaturated oxygenated products [ 54 ] identified in the chloroform fraction of the Suaeda vermiculata contributed to the higher antioxidant value of the chloroform fraction.…”

“…Furthermore, the pheophytin-a, which was reported for the mild antioxidant activity, and isolated previously form Suaeda vermiculata chloroform fraction [ 48 ] as part of the ongoing study, also played a role in the antioxidant activity of the chloroform fraction. Additionally, the steroidal constituent, β-sitosterol [ 48 ], along with other unsaturated oxygenated products [ 54 ] identified in the chloroform fraction of the Suaeda vermiculata contributed to the higher antioxidant value of the chloroform fraction. Nonetheless, the mother liquor aq.-ethanolic extract was pursued in further studies into the biological evaluations as a hepatoprotective extract, since the traditional practice of the use of plants’ aerial parts as a whole for liver disorders provided an equivalence between the prevailing practice and the aq.-ethanolic extract, the mother liquor obtained from the plant’s extraction.…”

“…Notably, the presence of higher concentrations of antioxidant compounds, measured as radical scavenging capacity in DPPH and ABTS assays is a direct indication of the antioxidant potential of the plant’s constituents, but not necessarily the high presence of phenolics and flavonoids, which can only be the reasons for higher antioxidant potential, though these two categories of products, i.e., phenolics and flavonoids, seem to be the reason for the dominating antioxidant potential of the plant. The higher antioxidant presence in the extract and the fractions of the Suaeda vermiculata can be attributed to the high-salinity habitat of the plant in comparison to its other counterpart species growing in different climatic locations [ 43 ]; the former has been the herb of choice for the local population for treatment of liver disorders, especially jaundice [ 48 , 55 ]. However, the biomechanistics connections between the higher antioxidant concentration levels of compounds in the plant, and its hepatoprotective action have not been conclusively established, though it has been well-observed, and a relationship is proposed at the primary levels ( Figure 5 ), albeit without intricate and basic biomechanistics roles playing, and interconnections, or the feedback responsibilities of the components in the presented cycle.…”

“…The comparatively higher concentrations of the flavonoids and phenolics in the halophytes, in comparison to the other normal-conditioned plants, might also be the scientific explanation for their liver-protecting activity [ 43 , 48 , 54 , 81 , 82 ]. Moreover, the hypoglycemic effects reported from other plants of the genus Suaeda , for example, Suaeda fruticosa produced a significant decrease in the blood glucose levels in normal rats, and a further reduction in the diabetic rat models was observed [ 83 ].…”