Comparison of Phytochemical Content and Toxicity of n-hexana Extracts and Fractions of Padina australis

Renovascular hypertension was produced in Wistar-King strain female rats by left renal artery clipping. The clipped kidney and the contralateral untouched kidney were cut into cortical, corticomedullary and medullary portion. The cortical extract was divided into subcellular fractions by the stepwise centrifugation method. Enzyme activity of acid phosphatase, cytochrome C oxidase and glucose-6-phosphatase was determined to ascertain the nomenclature of each fraction.Renin content in these fractions was determined by bioassay and it was expressed as equivalent activity of angiotensin II per mg of tissue protein.Renin content in whole kidney extract of the clipped kidney was 2 times higher than that in the normal kidney described in our previous report. Cortex had also high renin content and it was 2 times higher than in the normal kidney, while medulla did not show any renin activity as well as that in the normal kidney. The renin content was highest in lysosomal fraction, which was 4 times greater than that in the normal kidney. Mitochondrial, mitochondrialmicrosomal or soluble fraction had also higher renin content than respective fraction in the normal kidney. No renin activity was found in microsomal fraction as well as in that fraction in the normal kidney.In the contralateral untouched kidney, renin activity could not be demonstrated in any of the extracts or subcellular fractions.

Antioxidants are compounds that inhibit free radicals. Many plants have antioxidant activity because they contain antioxidant compounds such as flavonoids. This study aimed to evaluate the antioxidant activity, phytochemical contents, total flavonoid content and to establish the relationship of total flavonoid content and antioxidant activity of extract and fractions of walnut (Canarium indicum L.) leaves. Antioxidant activity of ethanolic extract, n-hexane fraction, ethyl acetate fraction, and ethanolic-water fraction of walnut leaves was examined by using the DPPH method and determination of total flavonoid content was determined spectrophotometrically. The results showed that antioxidant activity of ethanolic extract, n-hexane fraction, ethyl acetate fraction, and ethanolic-water fraction based were 25.294 ± 0.055; 175.245 ± 0.4999; 20.135 ± 0.009; and 28.806 ± 0.0424 μg/ml, respectively. The phytochemical content of ethanol extract of walnut leaves are saponins, flavonoids, tannins, and polyphenols. The total flavonoid content of ethanolic extract, n-hexane fraction, ethyl acetate fraction and ethanolic-water fraction were 2.624 ± 0.012; 0.499 ± 0.023; 3.846 ± 0.006; and 1.596 ± 0.006 gram quercetin equivalent/gram extract, respectively. Correlation between antioxidant activity and total flavonoid content revealed that 63.2 % of antioxidant activity was influenced by the presence of flavonoid compounds.

Vertical Zinc Migration in Various Soil TypesIn this contribution the zinc content in selected soil types and soil reaction on zinc loading in kinetic model conditions with knock-down columns filled with soil samples are evaluated. The total zinc content after their draining, finish and decomposition by HF + HClO4mixture and its fractions content in extract of 2 mol dm-3HNO3, in extract of 0.05 mol dm-3EDTA and in extract of 0.01 mol dm-3CaCl2using atomic absorption spectrometry method were determined.The knock-down columns filled with soil samples in model kinetic conditions were used for observation of zinc sorption measure. The maximal water capacity of soils was determined and then solution of 280 mg zinc (ZnSO4.2H2O) per kilogram of soil was applied. The zinc contents in extraction solutions 2 mol dm-3HNO3, 0.05 mol dm-3EDTA and 0.01 mol dm-3CaCl2in drained 0.05 m high soil columns by atomic absorption spectrometry method were determined.The obtained results were evaluated by mathematical-statistical methods - multiple range analysis and linear regression. Achieved data were compared to allowed limit values.The results show different behavior of individual soil types against zinc loading. The soils showed different properties, where zinc migration to the lower column layers was determined. The accent is given to zinc dynamics in neutral and acid soils. The limit value A (140 mg kg-1) was observed in Luvic Cambisol. The limit value A1 (40 mg kg-1) was observed in Eutric Regosol and Luvic Cambisol in upper layer of soil in column. The obtained results show high mobility of zinc in tested soils and thus its risk for ecosystems.

Hypertension is still one of the biggest health problems in the world, including in Indonesia. The high prevalence rate has encouraged a lot of research to find antihypertensive drugs and other alternative healing methods, especially those using natural ingredients. One plant that has antihypertensive activity is the roselle flower (Hibiscus sabdariffa L.). This research aimed to observe the inhibitory activity of roselle flower extract on the angiotensin-converting enzyme (ACE), determine the potential toxicity of roselle flower extract on shrimp larvae (Artemia salina Leach), and determine the phytochemical content in it. Roselle flower simplicia was extracted using the maceration method using a 70% ethanol solvent. The extract obtained was then tested for its inhibitory activity against ACE photometrically using a microplate reader. The observed inhibitory activity was calculated in terms of IC50. The toxicity of the extract was determined by the brine shrimp lethality test (BSLT) to see how toxic the roselle flower extract was to shrimp larvae. The secondary metabolite content in the extract was determined qualitatively and quantitatively. The research results showed that the extraction yield obtained was 32,63%. The IC50 value of roselle flower extract against ACE was 295,36 ppm. The toxicity test on shrimp larvae showed that the LC50 value obtained was 334,02 ppm. The results of qualitative phytochemical tests show that roselle flower extract contained flavonoids, quinones, and steroids. The flavonoids and phenolic content in roselle flower extract were 0,42% and 0,91%, respectively. Based on these results, the phytochemical content of roselle flower extract inhibited ACE activity, and its compounds can be used as ingredients for developing hypertension drugs

Scrophularia striata is a perennial plant which is native in all parts of Iran, Turkey, and Azerbaijan. In this study, the total phenol content, antioxidant and larvicidal activities of total extract and different fractions of this plant were evaluated. The aerial parts of S. striata were collected from Boli village, Illam Province, western Iran in Apr 2013. The total phenol content of total extract and different fractions were evaluated by Folin-Ciocalteu method. Moreover, antioxidant activity was tested by DPPH and FRAPS assays. Larvicidal activity was investigated according to standard method described by WHO. Ethyl acetate fraction (EF) had the highest content of total phenol (75.9±0.06mg Gallic acid equivalent/g dry extract). Furthermore, among the tested extract, methanol-water fraction (MWF), total methanol extract (TME) and water fraction (WF) showed the highest antioxidant activity in the DPPH assay (IC50= 226.8, 283.66 and 299.4 μg.ml-1, respectively). In FRAP assay MWF and WF and TME had the highest antioxidant activities (664.4±0.002, 565.3±0.003, 519.5±0.003mmol FeII/g dry extract, respectively). Ethyl acetate fraction had maximum larvicidal activity (LC50 49.1ppm) followed by TME (LC50 64.26ppm) and hexane fraction (HF) (LC50 89.69). Scrophularia striata collected from west of Iran illustrated considerable antioxidant and larvicidal effects and further in vitro and in vivo experimental models for investigation would be required.

The purpose of this study was to analyze the extraction efficiency of antioxidants from mango peel by comparing two techniques: microwave-assisted (MAE) and traditional solvent (TE) extraction. The number of extraction steps, water content in the extractant, peel weight-to-solvent volume ratio in extractions and extraction time all had an influence on obtaining extracts with high antioxidant capacity, but the extraction technique and the water content in the extractant were the factors with the greatest effect. Using three steps, a water content of 50 % in the ethanol:water extractant, an extraction time of 60 min and a weight-to-volume ratio of 1:10 or 1:50 (w/v) led to the highest antioxidant activity and phytochemicals content in extracts. The extraction time needed to extract phytochemicals from mango peel was similar when MAE and TE were used. However, the antioxidant capacity and phytochemical content were around 1.5-6.0 times higher in the extracts obtained by MAE.

Aim: The aim of this work is to evaluate the in ‐vitro antioxidant activity as well as estimate the total phenolic and flavonoid content of methanolic extract and fractions of Cleome viscosa L. Study Design: Evaluation of the antioxidant capacity of Cleome viscosa L. Methodology: Antioxidant activity of the methanol extract (MCV) and derived hexane (HCV) and ethyl acetate (ECV) fractions of Cleome viscosa L. was performed by DPPH radical scavenging, Ferric reducing antioxidant power (FRAP) assay and phosphomolybdate assay. The total phenolic and total flavonoid contents were also estimated using Folin-Ciocalteu and colorimetric aluminum chloride methods respectively. Results: The ethyl acetate fraction showed the highest total phenolic content (77.33±4.21 mg of gallic acid equivalent/g of extract (r 2 =0.9646)) and total flavonoid content (71.677±8.125 mg of quercetin equivalent/g of extract (r 2 =0.9449)). In the DPPH radical scavenging test, the IC 50 value of MCV, ECV and HCV was 0.91, 0.34 and 1.06 mg/mL, respectively also showing ethyl acetate

Spiny burs of Castanea mollissima Blume (Chinese chestnut) are usually discarded as industrial waste during post-harvesting processing. The objective of this study was to establish an extraction and isolation procedure for tannins from chestnut burs, and to assess their potential antioxidant activity. Aqueous ethanol solution was used as extraction solvent, and HPD 100 macroporous resin column was applied for isolation. The influence of solvent concentration in the extraction and elution process on extraction yield, tannins and polyphenols content, as well as antioxidant potential, including DPPH and ABTS radical scavenging ability, reducing power ability and cellular antioxidant ability were assessed. In both the extraction and isolation process, 50% aqueous ethanol led to superior total tannins and polyphenols content as well as significantly higher antioxidant activity. In addition, the antioxidant activity and the total tannins content in extracts and fractions had a positive linear correlation, and the predominant components responsible for antioxidant activities were characterized as hydrolysable tannins. To the best of our knowledge, this is the first report on the enrichment of tannins from burs of C. mollissim using macroporous resin chromatography, and to assess the cellular antioxidant activity of them.

Quantification of phenolic contents and antioxidant capacity of Atlantic sea cucumber, Cucumaria frondosa

Indonesia is an archipelagic country with a very wide coastal area. Lombok Island has a coastal area with quite extensive mangrove forest areas. Rhizopora mucronata was the most dominant species found in the region. The Rhizopora mucronata species was a plant from the Rhizoporaceae family reported to have antibacterial, antifungal, and antioxidant activity. These biological activities were caused by the presence of secondary metabolite compounds in R. mucronata, one of which is flavonoids. Research regarding the flavonoid content of R. mucronata leaves has not been explored. Therefore, the study aimed to determine the total flavonoid content in extracts and fractions of R. mucronata leaves using the spectrophotometric UV-Vis method. R. mucronata mangrove leaves simplicia were extracted using the sonication method using 96% ethanol solvent, then fractionated using the liquid-liquid extraction method with n-hexane, ethyl acetate, and water. The percentage yield of 96% ethanol extract, water fraction, ethyl acetate fraction, and n-hexane fraction respectively were 42.33%; 43.147%; 40.49%; 9.74%; and 32.49%. The TLC test showed the presence of flavonoid compounds detected in the extract and fractions which were marked with blue spots. The total flavonoid contents in the 96% ethanol extract, water fraction, ethyl acetate fraction, and n-hexane fraction were respectively (12,980; 14,160; 23,880; and 25,350) mg Quercetin equivalent/gram sample (mg QE/ g). The total flavonoid content of the extract and fractions of R. mucronata leaves were analyzed using the One-way ANOVA test method. The total flavonoid content in the n-hexane fraction and ethyl acetate fraction was significantly higher than the 96% ethanol extract and water fraction. The total flavonoid content of R. mucronata leaves was relatively high, which contributes to several of its biological activities.

The stem bark and wood of Berberis aristata DC (Daruharidra) are one of the principal ingredients of traditional skin lighting and exfoliating scrub preparation in India. The standardised extract of B. aristata was screened to evaluate their in vitro antityrosinase activity and inhibition kinetics. Phytochemical and pharmacological studies were carried out with different solvent fractions of the methanol extract of B. aristata (MEBA). RP-HPLC analysis was used to determine the berberine content in extract and fractions of B. aristata. MEBA showed maximum berberine content. Extract and fractions of B. aristata contain the maximum amount of alkaloids than other constituents. In tyrosinase inhibition assay, MEBA was found to possess highest dose-dependent monophenolase and moderate diphenolase activity. The enzyme kinetic study revealed that MEBA possessed mixed type inhibition of monophenolase activity of tyrosinase. These bioactivities indicate that the MEBA has antihyperpigmentation potential in human skin.

On Timor island, Nusa Tenggara Timur, faloak barks (Sterculia quadrifida R.Br.) has been used empirically to restore stamina. Faloak bark ethanolic extract proved to have immunomodulatory activity in vitro, which can increase macrophage phagocytosis activity. This research aimed: (i) to determine the immunomodulatory active fraction of faloak bark ethanolic extract, (ii) to determine the total flavonoid contents of faloak extract and fractions, and (iii) to evaluate the correlation of the total flavonoid contents of those extract and fractions with their macrophage phagocytosis activity. The simplisia powder is macerated with 96% ethanol. The extract was dissolved in methanol:water (9:1v/v) was then subsequently partitioned with n-hexane, ethyl acetate, and water to obtain n-hexane fraction, ethyl acetate fraction, water fraction, and insoluble fraction. Faloak extract and fractions at concentration 62,5; 125; 250; 500μg/mL were tested for their effect on the peritoneal macrophage phagocytosis of Balb/c mice in vitro by the latex beads method. Phagocytosis capacity and phagocytosis index were analyzed using one-way anova and post hoc Tukey HSD test with 95% confidence level. The results showed that ethyl acetate fraction had the highest macrophage phagocytosis capacity and the highest total flavonoid content compared to other fractions. The highest macrophage phagocytosis capacity of ethyl acetate fraction at concentration of 250 μg/mL was 51,94±4,67%, this value was significantly different from cell control (7,50±1,29%), negative controls of 0,0625% dimethylsulphoxide (6,25±0,36%), as well as positive control of 200 μg/mL echinaceae extract syrup® (9,97±0,33%). The total flavonoid content of ethyl acetate fraction determined by aluminum chloride method was 4,290±0.029 mg of quercetin equivalent/g fraction. There was a positive and strong correlation between the total flavonoid content of these extract and fractions with their macrophage phagocytosis capacity (Pearson correlation coefficient of 0,781) and showing linear relationship y=4,721x+19,663; R2=0,61.

Anti-tyrosinase and antimelanogenic effect of cinnamic acid derivatives from Prunus mahaleb L.: Phenolic composition, isolation, identification and inhibitory activity.

Phenolic compounds are one of the main parts in secondary metabolites. Parijoto fruit is a family of Melastomataceae which contains flavonoids as phenolic compounds which are known to have antioxidant potential. Flavonoids have an-OH groups that play an active role as free radical scavengers. The study was conducted as initial screening in testing the antioxidant activity of extracts and fractions of the Parijoto fruit (Medinilla speciosa). The study began with the phytochemical screening process using the Thin Layer Chromatography method to ensure the presence of flavonoid content in extracts and fractions of parijoto fruit, then proceed with the measurement of antioxidant power. Testing the antioxidant activity using the ABTS method (2,2 azinobis (3-ethylbenzothiazolin) -6-sulfonic acid) which is specific, simple and easy to apply. Parijoto fruit extracts and fractions were made in a series of concentrations, measured antioxidant activity, and each IC50 value was determined. Phytochemical screening test results showed the presence of flavonoids in extracts and fractions of parijoto fruit. The results of measurements of antioxidant power in extracts and fractions of parijoto fruit gave IC50 values of 6,520 ppm in ethanol extracts with very strong antioxidant categories. The n-hexane fraction produced an IC50 value of 118,424 ppm with a moderate antioxidant category, while the IC50 value of ethyl acetate and ethanol fractions was 4,246 ppm and 3,874 ppm, respectively, with a very strong antioxidant category. The activity and value of IC50 produced by extracts and fractions of Parijoto fruits showed potential as a candidate of antioxidant.Keywords: ABTS, Antioxidant, Flavonoids, Medinilla speciosa, Phenolic

Jackfruit (Artocarpus heterophyllus Lamk) is a plant from the Moraceae family that is widespread in Indonesia. Empirically, jackfruit can be used to cure hypertension, diabetes, cancer, asthma, dermatosis, coughs, wounds, acne, and diarrhea. The bioactive compounds in jackfruit include phenolics and flavonoids, which function as natural antioxidants. This research investigated the antiradical activity of jackfruit seed extracts and fractions using DPPH (2-2-diphenyl-1-picrylhydrazyl) and examined the total phenolic and flavonoid contents which may be developed as functional medicines and foods. The jackfruit seed powder was extracted using the maceration method. Radical scavenging activities were measured using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method. Total phenolic contents were determined using the Folin-Ciocalteu method. Meanwhile, the contents of flavonoids were determined using the aluminum chloride complex colorimetric method. The IC50 value of ethyl acetate fraction, methanol extract, n -hexane fraction, and water fraction was 5.435±0.064 µg/mL, 5.639±0.302 µg/mL, 7.201±0.475 µg/mL, and 9.134±0.2911 µg/mL, respectively. The phenolic and flavonoid contents of ethyl acetate fraction, methanol extract, n-hexane fraction, and water fraction were 49.597±1.589, 47.949±1.966, 41.214±4.354 and 35.504±0.913 g GAE/100 g of sample, respectively for phenolic, and 70.199±0.458, 65.228±0.615, 59.907±0.719, and 54.234±0.351 g of quercetin equivalent/100 g of sample, respectively of flavonoids, with a correlation value (R2 ) to antiradical power of 0.4582 for phenolic and 0.5281 for flavonoids. The ethyl acetate fraction of jackfruit seeds can be further developed as an anti -radicals and functional food.